CN113797338B - DBC1 for regulating cell aging and application thereof - Google Patents
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Abstract
The invention discloses DBC1 for regulating cell senescence and application thereof. The invention provides an application of a DBC1 promoter in preparing a medicament for preventing or treating diseases or symptoms related to aging. The invention also provides application of ML216 in preparation of a medicine for preventing or treating diseases or symptoms related to aging. The invention provides a new way for preventing or treating the diseases or symptoms related to the aging.
Description
Technical Field
The invention belongs to the field of biological medicines, and particularly relates to DBC1 for regulating cell senescence and application thereof.
Background
Cellular senescence refers to the shortening of telomeres by cell replication or external stress, manifested as cell cycle arrest. Cell senescence is mainly manifested in two forms, the first is that after cells are replicated for several times, telomeres are shortened, the cells are not proliferated any more, and a cell growth arrest state appears. The second is that cellular senescence can be induced by DNA damage or activation of aberrant oncogenes. Although senescent cells exhibit growth arrest, they are metabolically active and are characterized by distinct molecular and cellular changes, including cell cycle arrest, morphological changes, activation of pathways associated with p53 and p16 senescence, changes in chromatin structure, and significant changes in secreted factors such as cytokines and chemokines.
Although aging is inevitable and irreversible, the process and rate of aging can be intervened manually. Common methods of intervention in cellular senescence include:
1. preparing hypoxia state or adding active molecule and polyphenol medicine molecule to delay cell aging. For example, Eom et al use FGF-2, FGF-4, EGF, HGF growth factors to maintain the sternness of mesenchymal stem cells (see Young, Woo, Eom, et al. the role of growth factors in the main of bone in bone ground-derived mesenchymal stem cells [ J ]. Biochemical and Biophysical Research Communications,2014,445(1): 16-22.). Li Peng Cheng et al utilize a hypoxic environment to enhance the dryness of glioblastoma cancer stem cells (see Li P C, Zhou C, Xu L, et al. Hypoxia enhances stem of cancer stem cells in gliobastomama: An in vitro study [ J ]. International Journal of Medical Sciences, 2013,10(4): 399-.
2. The microcarrier is used for realizing three-dimensional culture of cells, namely a three-dimensional scaffold or gel is prepared to provide a proper growth environment for the cells, so that the effect of delaying cell aging is achieved. For example, Wei J et al successfully maintained the activity of mouse embryonic stem cells by the preparation of three-dimensional hydrogel scaffolds (see Wei J, Han J, ZHao Y, et al, the animal of the animal-dimensional scaffold on the structural of mouse embryonic stem cells [ J ]. Biomaterials, 2014,35(27): 7724-7733.).
However, these methods have some drawbacks, such as: the direct addition of active molecules often results in poor drug controllability, and the three-dimensional culture method has the problems of low collection efficiency and the like. Therefore, there remains a need in the art to find new agents that inhibit cellular aging.
Disclosure of Invention
In view of the defects of the prior art, the invention aims to provide a medicament for inhibiting cell aging, and the invention adopts the following technical scheme based on the aim:
in a first aspect, the present invention provides a pharmaceutical composition comprising an enhancer of DBC 1.
Further, the promoter includes an expression vector containing DBC1 or a recombinant protein of DBC 1.
In a second aspect of the invention, there is provided an agent for facilitating the interaction of DBC1 with BLM, said agent comprising ML 216.
In a third aspect, the present invention provides a method for screening a candidate drug for preventing or treating a disease or disorder associated with aging, the method comprising the steps of:
(1) treating a system expressing or containing DBC1 with a test substance;
(2) detecting the expression level of DBC1 in the system;
(3) selecting the substance capable of up-regulating the expression level of DBC1 as a candidate drug.
Further, the aging-related diseases or disorders include lung diseases and disorders, kidney diseases and disorders.
As an embodiment, the pulmonary diseases and disorders include pulmonary fibrosis, chronic obstructive pulmonary disease, asthma, cystic fibrosis, emphysema, bronchiectasis, and pulmonary diseases with age-related loss of lung function, preferably pulmonary fibrosis.
As an embodiment, the renal diseases and conditions include renal dysfunction, renal fibrosis, renal failure.
In a fourth aspect, the invention provides the use of any one of:
(1) use of a pharmaceutical composition according to the first aspect of the invention or an agent according to the second aspect of the invention in the manufacture of a medicament for inhibiting cellular senescence;
(2) use of a pharmaceutical composition according to the first aspect of the invention or an agent according to the second aspect of the invention in the manufacture of a medicament for the prevention or treatment of a disease or condition associated with aging;
(3) use of DBC1 in screening for a candidate drug for preventing or treating a disease or disorder associated with aging.
Further, the cellular senescence is DNA damage-induced cellular senescence.
Further, the cell is a mammalian cell.
Further, the mammalian cells include lung cells and kidney cells.
Further, the lung cell is a lung fibroblast.
Further, the lung fibroblast is a human embryonic lung fibroblast.
Further, the human embryonic lung fibroblast is IMR-90.
Further, the kidney cell.
Furthermore, the kidney cell is a human embryonic kidney cell.
Furthermore, the human embryonic kidney cells comprise 293 cells, 293T cells and 293FT cells.
Furthermore, the human embryonic kidney cell is 293T cell.
Further, the aging-related diseases or disorders include lung diseases and disorders, kidney diseases and disorders.
As one embodiment, the pulmonary diseases and disorders include pulmonary fibrosis, chronic obstructive pulmonary disease, asthma, cystic fibrosis, emphysema, bronchiectasis, and age-related pulmonary loss of function, preferably pulmonary fibrosis.
As an embodiment, the kidney diseases and conditions include renal dysfunction, renal fibrosis, renal failure.
Further, the medicine also comprises a pharmaceutically acceptable carrier and/or an auxiliary material.
Furthermore, the dosage forms of the medicine comprise tablets, capsules, granules, pills, dripping pills, syrups, powders, suppositories, drops, emulsions, injections, solutions or suspensions.
The invention has the following advantages:
the invention provides application of a promoter of DBC1 in preparing a medicament for preventing or treating diseases or symptoms related to aging.
The invention also provides application of ML216 in preparing a medicament for preventing or treating diseases or symptoms related to aging.
The invention also provides a method for screening a candidate drug for preventing or treating the aging-related disease or disorder.
Drawings
FIG. 1 is a graph showing the results of experiments demonstrating that the knockdown of DBC1 promotes senescence and inhibits apoptosis against DNA damage, wherein, in graph A, the results of experiments for analyzing Cell viability of DBC1 gene-knocked-down cells treated with cisplatin (50. mu.M) using a Cell Counting Kit (CCK-8), and the expression level of DBC1 gene in DBC1 gene-knocked-down cells are shown in the right graph; panel B is a graph showing the results of an experiment using annexin V staining and flow cytometry to quantify the level of apoptosis in DBC1 gene knock-down cells treated with staurosporine (1 μ M,2 hours); FIG. C is a graph showing the results of experiments in which the DBC1 gene knockdown cells treated with etoposide (50. mu.M, 24 hours) were analyzed for BCL-2, p53, and p21 protein levels using Western Blot; panel D is a graph of the results of cell cycle analysis performed on etoposide (10 μ M,24 hr) treated DBC1 gene knock-down cells; FIG. E is a graph of SA- β -Gal staining analysis experimental results and statistical analysis of DBC1 gene knock-down IMR-90 cells induced by etoposide or dimethylsulfoxide (control); FIG. F is a statistical graph of protein levels for detection of IL-6, IL-8, IL-1 α, CXCL-1, CXCL-2, MMP-3 and MMP-9 in DBC1 gene knockdown cells using RT-PCR analysis; FIG. G is a graph of experimental results and a graph of statistical analysis of proliferation of DBC1 gene knockdown cells analyzed by Ki-67 staining after etoposide-induced cellular DNA damage.
Fig. 2 is an experimental graph demonstrating that DBC1 modulates senescence induction by preserving BLM during DNA damage, wherein panel a is a graph of experimental results of Western Blot analysis of BLM, p16, and p21 protein levels in etoposide (50 μ M,24 hr) -treated DBC1 gene knock-down cells; panel B is a graph of the results of Western Blot analysis of DBC1, BLM, p16, and p21 protein levels in etoposide (50 μ M,24 hr) treated BLM gene knock-down cells; panel C is a graph of the results of cell cycle analysis of BLM knockdown cells; FIG. D is a graph of the results of SA- β -Gal staining analysis of BLM knockdown IMR-90 cells after DIS induction by etoposide and statistical plots; FIG. E is a graph showing the results of experiments in which BLM overexpressing DBC1 was knocked down for levels of BLM and p21 protein in cells using Western Blot analysis after DNA damage induced by etoposide (50. mu.M, 24 hours); FIG. F is a graph of SA- β -Gal staining analysis experimental results and statistical analysis of BLM gene knockdown IMR90 cells overexpressing DBC1 after DIS induction by etoposide.
FIG. 3 is an experimental graph showing a similarity in senescence-associated transcription patterns between BLM-knocked-out cells and DBC 1-knocked-out cells by RNA sequencing analysis, in which graph A is a Venn diagram of a common differentially expressed gene in DBC 1-knocked-down cells and BLM-knocked-down cells; FIG. B is a graph showing the results of gene ontology analysis of differentially expressed genes commonly found in DBC1 gene-knocked-down cells and BLM gene-knocked-down cells; FIG. C is a graph showing the results of an experiment for analyzing the common differentially expressed genes of DBC1 gene-knocked-down cells and BLM gene-knocked-down cells using KEGG enrichment; panel D is a graph showing the results of experiments analyzing the effect of etoposide treatment (50. mu.M, 24 hours) on the expression levels of senescence-associated genes in DBC1 gene knock-out cells and BLm gene knock-out cells.
FIG. 4 is an experimental diagram demonstrating that DBC1 prevents BLM degradation by binding thereto and inhibits induction of p21 after DNA damage, wherein panel A is a diagram showing the results of experiments using Western Blot analysis for knocking down the levels of BLM and p21 protein in DBC1 gene-treated cells for 24 hours with etoposide (100 μ M) or Z-VAD (OMe) -FMK, panel B is a diagram showing the results of experiments using co-immunoprecipitation to detect the interaction of DBC1 with BLM, panel C is a diagram showing the results of co-immunoprecipitation of full-length or truncated DBC1 with BLM, bottom panels are a diagram showing the structures of full-length DBC1 and truncated mutants (FL represents full-length, δ Nudix represents deletion of Nudix domain 463, and δ NT represents deletion of N-terminal domain (residues 1-263); panel D is a graph of the results of experiments in which levels of BLM and p21 protein in DBC1 were knocked down in cells by Western Blot analysis after etoposide-induced (50. mu.M, 24 hours) cellular DNA damage.
FIG. 5 is an experimental graph demonstrating that ML216 protects BLM from degradation and inhibits DNA damage-induced senescence by promoting DBC1-BLM interaction, wherein, the graph A is a graph of the experimental result of detecting the influence of ML-216 on the DBC1-BLM interaction by adopting a co-immunoprecipitation experiment, panel B is a graph of the results of Western Blot analysis of BLM and p21 protein levels 24 hours after treating DBC1 gene knock-down cells with etoposide (100. mu.M) or ML216 (50. mu.M), panel C is a graph of the results of in vitro immunoprecipitation experiments with DBC1 and BLM at various concentrations of ML216, FIG. D is a graph showing the results of experiments using Western Blot to detect the levels of BLM, gamma H2AX and p21 protein in 293T cells treated with different concentrations of ML216 (24 hours), FIG. E is an experimental graph of the number of SA- β -Gal positive cells detected after co-treatment of IMR90 cells DIS induced by etoposide or etoposide with different concentrations of ML 216; FIG. F is a statistical plot of the number of SA- β -Gal positive cells detected after co-treatment of IMR90 cells DIS induced with etoposide or etoposide with different concentrations of ML 216; FIG. G is a statistical graph of expression levels of p16, p21, IL-1 alpha, IL-6, CXCL-1 in IMR90 cells after induction of DIS by RT-PCR; FIG. H is a graph showing the results of experiments to determine the repair efficiency of Homologous Recombination (HR) in 293T cells treated with different concentrations of ML216 for 24 hours; FIG. I is a graph showing the results of an experiment for measuring the HR repair efficiency of DBC1 gene knockout 293T cells treated with ML216 (50. mu.M at 24 hours).
FIG. 6 is a graph demonstrating that ML-216 reduces fibrosis and improves lung function in IPF mice, wherein graph A is a schematic design of bleomycin induced pulmonary fibrosis in C57BL/6J mice; panel B is a statistical plot of lung function parameter measurements in mice; panel C is a representative plot of Masson staining of mouse lungs; FIG. D is a graph showing the results of an experiment for measuring the collagen content in the lung tissue of a mouse by hydroxyproline assay; panel E is a statistical graph of mRNA expression levels of the fibrosis marker genes COL1A1, MMP2, FN1, TGF-. beta.and CTGF in mouse lungs.
FIG. 7 is a graph showing that ML-216 can delay cellular senescence at the lung molecular level in IPF mice, in which graph A is a statistical graph of the protein levels of SASP factors (IL-6, MCP-1, CXCL-1) in the lungs as measured by ELISA; FIG. B is a statistical chart of protein levels of SASP factors (IL-6, MCP-1, CXCL-1) in serum measured by ELISA; FIG. C is a graph showing the results of an experiment for analyzing the level of p21 protein in mouse lung by Western Blot and a statistical analysis graph; panel D is a representative image of mice lung aging levels analyzed using SA- β -Gal staining; panel E is a quantitative statistical plot of mouse lung SA- β -Gal positive cells.
FIG. 8 is a graph showing the effect of DBC1 overexpression on Cell survival rate and apoptosis level, wherein, graph A is a graph showing the result of Cell survival rate analysis experiment using Cell Counting Kit (CCK-8) on cisplatin (50. mu.M) -treated cells overexpressing DBC1 gene, and DBC1 overexpression level is shown in the right graph; panel B is a representative scatter plot and quantitative analysis statistical plot of levels of apoptosis of DBC1 over-expressed cells treated with staurosporine (1 μ M,2 hours) using annexin V staining and flow cytometry.
Detailed Description
Accelerator for DBC1
The invention provides a pharmaceutical composition, which comprises an accelerant of DBC 1.
The "DBC 1" refers to the Gene with Gene ID 57805.
The promoter of DBC1 refers to any substance that can increase the stability of DBC1 gene or expression product, up-regulate the expression of DBC1, increase the effective duration of DBC1, or promote the transcription of DBC1 gene, and can be used in the present invention as a substance useful for up-regulating the expression of DBC1 gene, thereby being useful for preventing or treating aging-related diseases or disorders.
The promoter comprises an expression vector containing DBC1 or a recombinant protein of DBC 1. In a preferred embodiment of the present invention, the promoter of DBC1 is an expression vector containing DBC 1. The expression vector usually further contains a promoter, an origin of replication, and/or a marker gene.
Methods well known to those skilled in the art can be used to construct the expression vectors required by the present invention. These methods include in vitro recombinant DNA techniques, DNA synthesis techniques, in vivo recombinant techniques, and the like. The expression vector preferably comprises one or more selectable marker genes to provide a phenotypic trait for selection of transformed host cells, such as kanamycin, gentamicin, hygromycin, ampicillin resistance.
In the present invention, the expression vector is various vectors known in the art, such as commercially available vectors, including plasmids, cosmids, phages, viruses, and the like. The expression vector can be introduced into the host cell by a known method such as electroporation, calcium phosphate method, liposome method, DEAE-dextran method, microinjection, viral infection, lipofection, or binding to a cell membrane-permeable peptide.
Age-related diseases or disorders
The present invention provides a medicament for treating a disease or disorder associated with aging. A senescence-associated disease or disorder may also be referred to herein as a senescent cell-associated disease or disorder. Aging-related diseases or disorders include, for example, cardiovascular diseases and disorders, inflammatory diseases and disorders, autoimmune diseases and disorders, lung diseases and disorders, ocular diseases and disorders, metabolic diseases and disorders, neurological diseases and disorders (e.g., neurodegenerative diseases and disorders); age-related diseases and disorders caused by aging; age-related diseases; skin diseases and disorders; and transplantation-related diseases and disorders. The prominent feature of aging is the gradual loss or deterioration of function that occurs at the molecular, cellular, tissue, and biological levels. Age-related degeneration causes well-recognized pathologies such as muscle atrophy, atherosclerosis and heart failure, osteoporosis, pulmonary valve insufficiency, renal failure, neurodegeneration (including macular degeneration, alzheimer's disease, and parkinson's disease), and many others.
In a preferred embodiment of the invention, the senescence-associated disease or disorder is a pulmonary disease or disorder, including but not limited to pulmonary fibrosis, chronic obstructive pulmonary disease, asthma, cystic fibrosis, emphysema, bronchiectasis, and age-related pulmonary loss of function, preferably pulmonary fibrosis.
In another preferred embodiment of the present invention, the senescence-associated disease or disorder is a renal disease or disorder, including renal dysfunction, renal fibrosis, renal failure.
Pharmaceutically acceptable carrier
The medicament for treating the disease or the symptom related to the aging also comprises a pharmaceutically acceptable carrier and/or an auxiliary material.
The term "pharmaceutically acceptable carrier" includes any and all solvents, diluents or other liquid vehicles, dispersing or suspending aids, surfactants, isotonic agents, thickening or emulsifying agents, preservatives, solid binders, lubricants and the like suitable for use in preparing the particular dosage form desired. Some examples of materials that can be used as pharmaceutically acceptable carriers include, but are not limited to, sugars such as lactose, glucose, and sucrose; starches, such as corn starch and potato starch; cellulose and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose, and cellulose acetate; powdered gum tragacanth; malt; gelatin; talc; excipients such as cocoa butter and suppository waxes; oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and soybean oil; glycols, such as propylene glycol; esters such as ethyl oleate and ethyl laurate; agar; buffering agents such as magnesium hydroxide and aluminum hydroxide; alginic acid; pyrogen-free water; isotonic saline; ringer's solution; ethanol and phosphate buffer, and other non-toxic compatible lubricants such as sodium lauryl sulfate and magnesium stearate, as well as coloring agents, releasing agents, coating agents, sweetening, flavoring and perfuming agents, preservatives and antioxidants may also be present in the composition, according to the judgment of the formulator.
In the present invention, the agent for preventing or treating the aging-related disease or disorder can be administered to the mammal by various methods well known in the art. Including but not limited to: subcutaneous injection, intramuscular injection, transdermal administration, topical administration, implantation, sustained release administration, and the like.
The effective amount of promoter of DBC1 or ML216 described herein may vary depending on the mode of administration and the severity of the condition to be treated, among other things. The selection of a preferred effective amount can be determined by one of ordinary skill in the art based on a variety of factors (e.g., by clinical trials). Such factors include, but are not limited to: the promoter of DBC1 or pharmacokinetic parameters of ML216 such as bioavailability, metabolism, half-life, etc.; the severity of the disease to be treated by the patient, the weight of the patient, the immune status of the patient, the route of administration, and the like. For example, divided doses may be administered several times per day, or the dose may be proportionally reduced, as may be required by the urgency of the condition being treated.
Cells
The term "cell" includes prokaryotic and eukaryotic cells. Examples of commonly used prokaryotic cells include Escherichia coli, Bacillus subtilis, and the like. Commonly used eukaryotic cells include yeast cells, insect cells, and mammalian cells. Preferably, the cell is a mammalian cell.
In one embodiment, the mammalian cell is a lung cell, preferably a lung fibroblast, and in a specific embodiment of the present invention, the lung fibroblast is a human embryonic lung fibroblast, including but not limited to WI-38, KMB17, IMR-90, MRC-5, 2BS, preferably IMR-90.
In one embodiment, the mammalian cells are kidney cells including, but not limited to, monkey kidney CVI cell lines (e.g., COS-7, ATCC CRL 1651); human embryonic kidney lines (e.g., 293T, 293 FT); baby rat kidney cells (e.g., BHK, ATCC CCL 10); monkey kidney cells (e.g., CVI ATCC CCL 70); vero kidney cells (e.g., VERO-76, ATCC CRL-1587); canine kidney cells (e.g., MOCK, ATCC CCL34), and the like.
Drug screening
The present invention provides a method for screening a candidate drug for preventing or treating an aging-related disease or disorder, the method comprising the steps of:
(1) treating a system expressing or containing DBC1 with a test substance;
(2) detecting the expression level of DBC1 in the system;
(3) selecting the substance capable of up-regulating the expression level of DBC1 as a candidate drug.
The system for expressing or containing DBC1 can be a cell (or cell culture) system, and the cell can be a cell endogenously expressing DBC 1; or may be a cell recombinantly expressing DBC 1. The system expressing or containing DBC1 can also be a subcellular system, a solution system, a tissue system, an organ system or an animal system (such as an animal model, preferably a non-human mammalian animal model, such as mouse, rabbit, sheep, monkey, etc.), and the like.
Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Further, some terms are explained as follows:
the term "and/or" as used herein in phrases such as "a and/or B" is intended to include both a and B; a or B; a (alone); and B (alone). Likewise, the term "and/or" as used in phrases such as "A, B and/or C" is intended to encompass each of the following embodiments: a. B and C; A. b or C; a or C; a or B; b or C; a and C; a and B; b and C; a (alone); b (alone); and C (alone).
The term "treatment" as used herein generally relates to the treatment of a human or animal (e.g., as applied by a veterinarian) wherein some desired therapeutic effect may be achieved, for example, inhibiting the development of a condition (including reducing the rate of development, halting development), ameliorating the condition, and curing the condition. Treatment as a prophylactic measure (e.g., prophylaxis) is also included. The use of a patient who has not yet developed a condition but who is at risk of developing the condition is also encompassed by the term "treatment".
The present invention is further illustrated below with reference to specific examples, which are intended to be illustrative only and are not to be construed as limiting the invention. Those of ordinary skill in the art will understand that: various changes, modifications, substitutions and alterations can be made to the embodiments without departing from the principles and spirit of the invention, the scope of which is defined by the claims and their equivalents. The following examples are examples of experimental methods not indicating specific conditions, and the detection is usually carried out according to conventional conditions or according to the conditions recommended by the manufacturers.
Example 1 Effect of DBC1 deletion on cells
The first experiment method comprises the following steps:
1. to examine the effect of DBC1 gene knock-down or overexpression on 293T Cell survival after DNA damage, Cell viability analysis was performed on DBC1 gene knock-down 293T cells (pRS-DBC1, Origene, TR303704) or DBC1 overexpression 293T cells (pMSCV-Flag-DBC1, self-constructed plasmid) treated with cisplatin (50. mu.M) using a viable Cell Counting Kit (Cell Counting Kit-8, CCK-8, jd 226). To examine the effect of DBC1 gene knockdown or overexpression on 293T cell Apoptosis, the level of Apoptosis of DBC1 gene-knockdown cells or DBC 1-overexpressing 293T cells treated with staurosporine (1 μ M,2 hours) was quantitatively analyzed using Annexin V staining (FITC Annexin V Apoptosis Detection Kit I, BD Biosciences, 556547) and flow cytometry. Western Blot is adopted to detect the expression level of the DBC1 gene in DBC1 gene knockdown cells or DBC1 overexpression 293T cells.
2. The DBC1 gene treated with etoposide (50. mu.M, 24 hours) was analyzed by Western Blot for knock-down of BCL-2, p53 and p21 protein levels in 293T cells.
3. Cell cycle analysis was performed on etoposide (10 μ M,24 hr) treated DBC1 gene knockdown 293T cells. Cells were seeded into 6-well cell culture plates and treated with 10 μ M etoposide for 24 hours. Cells were then harvested and washed twice with PBS. Cells were fixed at least one night in 70% pre-chilled ethanol at-20 ℃. The following day, the fixed cells were washed twice with PBS and stained in 50. mu.g/ml PI supplemented with RNase A (100ug/ml) for 30 min in the dark. The final cells were quantitatively analyzed by flow cytometry.
4. Etoposide, a DNA damaging agent, is used to induce cellular senescence, a process that is DNA damage-induced senescence (DIS). SA-beta-Gal staining analysis is carried out on the IMR-90 cells subjected to DBC1 gene knock-down induced cell senescence by etoposide, and the influence of DBC1 gene knock-down on IMR-90 cell senescence is researched, wherein the cells treated by DMSO are used as a control group. The DIS induction method comprises the following steps: early passage IMR-90 cells were treated with etoposide (20. mu.M) at 16 th passage for 24 hours. After 24 hours, the culture medium is replaced by fresh culture medium to replace cells, the cells continue to grow for 7 days, and the fresh culture medium is frequently replaced within 7 days, so that the IMR-90 cells are aged. SA- β -Gal staining was: cells were fixed in G/F fixative (PBS solution containing 50% glutaraldehyde and 37% formaldehyde) for 5 minutes at room temperature. After washing twice in PBS, the cells were placed in a staining solution [ containing 200mM potassium ferricyanide, 200mM potassium ferrocyanide, 200mM MgCl26M NaCl and 1mg/ml X-gal citrate/sodium phosphate buffer (aqueous solution containing 0.1M citric acid, 0.2M sodium phosphate)]No CO at 37 ℃2The humidifying chamber is placed for 2-4 hours. Finally, the film was observed under a microscope and photographed.
5. RT-PCR analysis detects the expression level of IL-6, IL-8, IL-1 alpha, CXCL-1, CXCL-2, MMP-3 and MMP-9 in DBC1 gene knock-down IMR-90 cells.
6. Ki-67 staining was performed on day 4 and 7 after etoposide-induced DNA damage to analyze proliferation of DBC1 gene knock-down IMR-90 cells, where DMSO-treated cells were the control group.
Second, experimental results
1. As shown in fig. 1A, knocking down the DBC1 gene can improve the cell survival rate after DNA damage;
2. as shown in fig. 1B, the number of cells undergoing apoptosis in the DBC1 gene knock-down cells was small;
3. as shown in fig. 8A, 8B, DBC1 overexpression increased cell mortality to cope with DNA damage, particularly increased levels of apoptosis.
4. As shown in fig. 1C, the decreased level of apoptosis in DBC1 gene knockdown cells following DNA damage was primarily attributed to the high level expression of the apoptosis inhibitor BCL-2, while the key pro-apoptotic factor p53 was not significantly altered.
5. p21 is a CDK inhibitor that prevents cell cycle progression through G1/S phase, and as shown in fig. 1D, DBC1 gene knock down cells showed prolonged G1 phase arrest following DNA damage as the level of p21 induction increased.
6. As shown in FIG. 1E, compared to the control group, in the IMR-90 cells with reduced DBC1, the number of senescence-associated galactosidase (SA-. beta. -gal) staining positive cells was increased 2-3 fold regardless of the presence or absence of DNA damage, and the expression levels of senescence-associated secreted phenotype (SASP) factors (e.g., IL-6, IL-8, MMP-3) were also significantly increased (as shown in FIG. 1F).
6. As shown in fig. 1G, Ki-67 staining results showed that the level of proliferation of DBC1 knockdown cells was reduced at day 4, day 7 after etoposide-induced DNA damage.
The above results demonstrate that inhibition of the expression level of DBC1 promotes DNA damage, which in turn promotes cellular senescence and reduces apoptosis.
Example 2 mechanistic study of DBC1 Regulation of DNA Damage-induced cellular senescence
First, experiment method
1. Western Blot analysis DBC1 gene treated with etoposide (50 μ M,24 hours) knockdown BLM, p16 and p21 protein levels in 293T cells. Western Blot analysis BLM gene treated with etoposide (50 μ M,24 hours) knockdown DBC1, BLM, p16, and p21 protein levels in 293T cells.
2. Cell cycle analysis was performed on BLM knockdown 293T cells.
3. After induction of DIS by etoposide, SA- β -Gal staining analysis was performed on BLM knockdown IMR-90 cells.
4. BLM overexpressing DBC1 was knocked down for BLM and p21 protein levels in 293T cells using Western Blot analysis after DNA damage was induced by etoposide (50 μ M,24 hours).
5. After induction of DIS by etoposide, SA- β -Gal staining analysis was performed on BLM gene knockdown IMR90 cells overexpressing DBC 1.
6. To further investigate the correlation between DBC1 and BLM and its function in senescence, RNA sequencing analysis was performed on DBC1 knockdown cells or BLM knockdown 293T cells, respectively. Differentially expressed genes common in DBC 1-knockdown cells and BLM-knockdown 293T cells were presented in the form of venn plots and subjected to gene ontology analysis, -log10 (P-value) as a function of classification, satisfying a P-value < 0.05. The genes were differentially expressed in common between the DBC1 gene-knockdown cells and the BLM gene-knockdown cells compared to control cells using KEGG enrichment analysis.
7. The effect of etoposide treatment (50 μ M,24 hours) on the expression levels of senescence-associated genes in DBC1 gene knockout 293T cells and BLM gene knockout 293T cells was analyzed.
Second, experimental results
1. As shown in fig. 2A, BLM protein levels decreased in DBC1 gene knock-down cells after induction of DNA damage compared to control cells, with a trend approximately inversely proportional to the increase in p 21.
2. As shown in fig. 2B, when DNA damage was induced, the expression level of p21 was increased in BLM gene knock-down cells, the cell cycle G1 phase was prolonged (as shown in fig. 2C), and the expression level of p53 or p16 was not changed, which was similar to that of DBC1 gene knock-out cells (as shown in fig. 1C and fig. 1D), compared to the control group.
3. As shown in fig. 2D, in the pattern of cellular senescence induced by DNA damage, the level of senescence was increased in BLM-deleted IMR-90 cells compared to control cells.
4. As shown in fig. 2E, after DNA damage was induced by transfection of DBC1 into BLM-deleted cells, more BLM protein was retained with more DBC1 present, and the level of p21 decreased.
5. In a pattern of DNA damage-induced cellular senescence, transfection of DBC1 into BLM-deficient IMR-90 cells reduced the number of SA- β -gal positive cells, inhibiting senescence levels (as shown in fig. 2F). Unlike DBC 1-knockdown cells (see FIGS. 1B and 1C) in which BCL-2 expression levels were elevated and apoptosis levels were reduced, knockdown of BLM had no effect on both BCL-2 expression levels and apoptosis levels.
6. RNA sequencing was performed on the DBC 1-knockdown cells and the BLM-knockdown cells, respectively, and transcripts with common changes in both sets of data were analyzed to obtain Gene Expression Profiles (GEPs) with 200 gene overlaps. Analysis of GO enrichment in Biological processes (Biological Process) indicates that "cell aging" is one of the most enriched upregulation pathways common to both datasets (as shown in FIG. 3B). KEGG analysis of DBC1 and BLM co-regulated genes showed that the most enriched pathways are associated with oxidative phosphorylation, huntington's disease, metabolic pathways, parkinson's disease and alzheimer's disease, all of which are diseases associated with aging that if defective in function would lead to aging (as shown in figure 3C). In this context, comparison of transcriptional analysis of the effect of knockdown BLM or DBC1 on marker senescence-associated genes revealed that they had very similar patterns of action under DNA damage, and that absence of either DBC1 or BLM promoted senescence (as shown in fig. 3D), providing evidence that BLM and DBC1 had functional overlap in regulating cellular senescence.
Taken together, the above results demonstrate that DBC1 regulates DNA damage-induced cellular senescence by maintaining BLM abundance.
Example 3DBC1 protection of BLM from degradation by binding to BLM
First, experiment method
1. To further examine whether the reduction in BLM protein levels in DBC1 gene-knocked-down 293T cells was associated with caspase, cells were treated with the pan-caspase inhibitor Z-vad (ome) -FMK. Western Blot analysis was performed on levels of BLM and p21 protein in DBC1 gene-knocked-down 293T cells treated with etoposide (100. mu.M) or Z-VAD (OMe) -FMK for 24 hours.
2. Co-immunoprecipitation was used to analyze the interaction of DBC1 and BLM.
3. A series of truncations were performed on DBC1 and full-length or truncated DBC1 was subjected to co-immunoprecipitation experiments with BLM. Schematic structures of full-length and truncated mutants of DBC1(pcDNA3.1-DBC1-V5, pcDNA3.1-DBC 1-V5. DELTA.NT, pcDNA3.1-DBC 1-V5. DELTA.Nudix, self-constructed plasmids) are shown in the bottom panel of FIG. 4C. FL represents full length; Δ Nudix denotes the deletion of the Nudix domain (residues 339-463); Δ NT denotes the deletion of the N-terminal domain (residues 1-263).
4. To test whether binding of DBC1 to BLM helped DBC1 protected BLM from cleavage, introduction of full-length (FL) or N-terminal truncation mutant DBC1(Δ NT) into DBC1 knockdown 293T cells, and levels of BLM and p21 protein in the cells were analyzed by Western Blot after etoposide-induced (50 μ M,24 hours) DNA damage.
Second, experimental results
1. As shown in FIG. 4A, at a concentration of 50. mu.M, Z-VAD (OME) -FMK reduced the loss of BLM after DNA damage to a level comparable to that of the control group, with a corresponding reduction in the level of p 21. This result demonstrates that the presence of DBC1 reduces the cleavage of BLM by DNA-damage activated caspases.
2. As shown in fig. 4B, DBC1 interacts with BLM. Further defining the domains in DBA1 that bind to BLM, experimental results showed that the DBC1 mutant lacking the N-terminal 243 amino acid residues was unable to interact with LM, whereas the Nudix domain in DBC1 was absent no effect on the interaction of DBC1 with BLM (as shown in fig. 4C).
3. As shown in fig. 4D, reintroduction of N-terminally truncated mutant DBC1 had no significant effect on BLM reduction compared to DBC1 gene knock-down cells, while reintroduction of full-length DBC1 inhibited BLM protein reduction due to DNA damage, similar to that of control cells. When full-length DBC1 was reintroduced, the induction of p21 by DNA damage was reduced, while the introduction of non-interacting mutants did not give similar results.
Taken together, the above results demonstrate that DBC1 protects BLM from DNA damage-induced cleavage by binding to BLM.
Example 4ML216 protects BLM from degradation by promoting DBC1-BLM interaction and inhibits DNA damage-induced senescence.
First, experiment method
1. Co-immunoprecipitation experiments were performed 24 hours after treating 293T cells with etoposide (100. mu.M), ML216 (50. mu.M), or both, and the effect of ML-216 on the DBC1-BLM interaction was observed.
2. DBC1 gene knockdown 293T cells were treated with etoposide (100. mu.M) or ML216 (50. mu.M) for 24 hours and then analyzed for BLM and p21 protein levels by Western Blot.
3. In vitro immunoprecipitation experiments of DBC1 with BLM were performed at different concentrations of ML 216. After 4 equal parts of lysates of 293T cells highly expressed by FLAG-DBC1 were incubated with 0, 20, 50 and 100. mu.M ML-216 for 12h, immunoprecipitation experiments were performed with FLAG antibody, and the level of BLM protein was detected by Western Blot.
4. Western Blot detects the protein levels of BLM, γ H2AX and p21 in 293T cells treated with different concentrations of ML216 (24 hours) with etoposide treated group as positive control (100 μ M,24 hours).
5. The number of SA-beta-Gal positive cells is detected after IMR90 cells DIS are induced by co-treatment of etoposide or etoposide and ML216 with different concentrations, and the expression levels of p16, p21, IL-1 alpha, IL-6 and CXCL-1 in the induced IMR90 cells are detected by RT-PCR.
6. 293T cells were treated with different concentrations of ML216 for 24 hours and the efficiency of cellular Homologous Recombination (HR) repair was examined. The HR step is as follows: the pcDNA3.1-HR plasmid (the self-constructed plasmid), the pCBASCEI plasmid (Addgene, 26477) and the dsRed2-N1 plasmid (Addgene, 54493) were transfected into cells. Drug treatment was performed 48 hours after transfection and the final cells were quantified by flow cytometry. The ratio of GFP positive cells to RFP positive cells was used to calculate HR repair efficiency.
7. DBC1 gene knock-down on the HR repair efficiency of 293T cells treated with ML216 (50. mu.M, 24 hours) was tested.
Second, experimental results
1. As shown in fig. 5A, after ML216 treatment, more BLM protein was pulled down by Flag-DBC1 regardless of the presence of DNA damage, and ML216 also increased BLM protein levels in the input group.
2. As shown in fig. 5B, ML216 increased BLM protein levels almost completely inhibiting p21 protein levels after etoposide-induced DNA damage in control cells, whereas BLM protein levels were less increased and p21 protein levels were only inhibited by about 50% in DBC1 knock-out cells, showing the functional dependence of ML216 on DBC 1.
3. As shown in fig. 5C, the results indicate a dose-dependent increase in the interaction of BLM and DBC1 with ML 216.
4. As shown in fig. 5D, the expression levels of senescence markers, including p21, γ H2AX, were not significantly changed in ML 216-treated cells. However, ML216 can reduce the levels of p21 and γ -H2AX in etoposide-induced DNA damage.
5. As shown in fig. 5E, 5F, in agreement with the results of downregulating senescence marker p21 and γ -H2AX protein levels, ML216 significantly reduced the proportion of SA- β -Gal positive senescent cells in DIS-induced cells by etoposide. As shown in FIG. 5G, ML216 also reduced the expression levels of SASP factors IL-1a, IL-6 and CXCL 1.
6. As shown in fig. 5H, ML216 processing increased HR repair efficiency, while when DBC1 was knocked down, ML216 processing did not increase HR repair efficiency (as shown in fig. 5I), which demonstrates that ML216 has a functional dependence on DBC 1.
Taken together, the above data demonstrate that ML216 protects BLM from degradation by promoting DBC1-BLM interactions, thereby inhibiting DNA damage-induced cellular senescence.
Example 5ML216 reduction of age-related pathological changes in Idiopathic Pulmonary Fibrosis (IPF) mouse model-Experimental methods
1. A single intratracheal instillation of 2mg/kg bleomycin induced pulmonary DNA damage and senescence in C57BL/6J mice (as shown in FIG. 6A).
2. One week after gavage, both groups of mice were injected intraperitoneally with ML217(0.5mg/kg) or vehicle 2 times a week for 3 weeks. Lung function parameters were measured in mice after 3 weeks, including: dynamic compliance, peak compliance, and chord compliance. The pulmonary fibrosis status was detected by Masson staining (Masson trichrome staining Kit, Solarbio, G1343) and H & E staining (H & E staining was performed by the department of histology in the key laboratory of the national institute of molecular biology of medicine of the kyoto institute of medicine). Hydroxyproline was used to determine the collagen content in mouse lung tissue (Hydroxyproline Assay Kit, Solarbio, BC 0255). Detecting the mRNA expression level of the mouse lung tissue fibrosis marker gene. ELISA was used to detect key SASP factors in Mouse lung and serum (Mouse CXCL1/KC (IL-8) ELISA Kit, Absin, ABS 520017; Mouse CCL2/JE/MCP-1ELISAKit, Absin, ABS 520016; Mouse IL-6ELISAKit, Absin, ABS 520004). Levels of pulmonary cell senescence in mice were analyzed using SA- β -Gal staining. For tissue SA- β -Gal staining, frozen sections were fixed in ice-cold fixative (pre-cooled PBS containing 2% formaldehyde and 0.2% glutaraldehyde) for 7 minutes. The samples were then washed 3 times with pre-cooled PBS for 10 minutes each. Frozen sections were surrounded with PAP pens and stained with staining solution (cocytotic staining solution) overnight in a humidified chamber without CO2 at 37 ℃. Finally, the film was observed under a microscope and photographed.
Second, experimental results
1. As shown in fig. 6B, bleomycin treatment reduced lung elasticity by 20-50% in mice, which was measured by compliance indices, such as dynamic compliance, peak compliance, and chord compliance, all of which were significantly improved after ML216 treatment.
2. As shown in fig. 6C, Masson staining, H & E staining results show that ML216 can reduce pulmonary fibrosis in IPF mice.
3. As shown in fig. 6D, the hydroxyproline assay results showed that the collagen content of the lung tissue of ML216 mice was significantly lower than that of the control group.
4. As shown in fig. 6E, the mRNA expression level of the pulmonary fibrosis marker gene was decreased in the ML216 group of mice.
5. Fibrosis of lung tissue significantly increased the protein levels of IL-6, CXCL1 and MCP-1 in the lung and serum. ML216 administration significantly reduced the protein levels of IL-6 in the lungs and MCP-1 in serum, while other factors were in a downward trend (as shown in FIGS. 7A and 7B). The induction rate of p21 in the lungs of the ML216 treated group was significantly lower than that of the control group (as shown in fig. 7C). As shown in fig. 7D and 7E, the lungs of the ML 216-treated mice were decreased in SA- β -Gal positive cells, which demonstrated that the overall senescence status of the ML 216-treated mice was lighter than that of the control mice.
Taken together, the above results demonstrate that ML216 administration can inhibit lung cell aging to reduce pulmonary fibrosis in IPF mice, thereby improving lung function.
The above description of the embodiments is only intended to illustrate the method of the invention and its core idea. It should be noted that it would be apparent to those skilled in the art that various modifications and variations can be made in the present invention without departing from the spirit of the invention, and these modifications and variations also fall within the scope of the claims of the present invention.
Claims (6)
1. Use according to any one of the following:
(1) use of an agent that promotes the interaction of DBC1 with BLM in the manufacture of a medicament for inhibiting cellular senescence;
(2) use of an agent that promotes the interaction of DBC1 with BLM in the manufacture of a medicament for the prevention or treatment of pulmonary fibrosis;
the reagent is ML216, and the reagent is,
the cells are lung fibroblasts.
2. The use according to claim 1, wherein the cellular senescence is DNA damage-induced cellular senescence.
3. The use of claim 1, wherein the lung fibroblast is a human embryonic lung fibroblast.
4. The use of claim 3, wherein the human embryonic lung fibroblast is IMR-90.
5. The use according to claim 1, wherein the medicament further comprises a pharmaceutically acceptable carrier and/or adjuvant.
6. Use according to claim 1, wherein the medicament is in the form of tablets, capsules, granules, pills, syrups, powders, suppositories, drops, emulsions, solutions or suspensions.
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| CN102985553A (en) * | 2010-04-15 | 2013-03-20 | 西特里斯药业公司 | Sirtuin activators and activation assays |
| WO2017211983A1 (en) * | 2016-06-09 | 2017-12-14 | Centre Hospitalier Et Universitaire De Montpellier | Methods using blm as a marker of multiple myeloma |
| WO2021084540A1 (en) * | 2019-10-30 | 2021-05-06 | Yeda Research And Development Co. Ltd. | Inhibitors of mmej pathway for prevention and treatment of pre-myeloid and myeloid malignancies |
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| CN102985553A (en) * | 2010-04-15 | 2013-03-20 | 西特里斯药业公司 | Sirtuin activators and activation assays |
| WO2017211983A1 (en) * | 2016-06-09 | 2017-12-14 | Centre Hospitalier Et Universitaire De Montpellier | Methods using blm as a marker of multiple myeloma |
| WO2021084540A1 (en) * | 2019-10-30 | 2021-05-06 | Yeda Research And Development Co. Ltd. | Inhibitors of mmej pathway for prevention and treatment of pre-myeloid and myeloid malignancies |
Non-Patent Citations (5)
| Title |
|---|
| DBC1的蛋白质结构与生物学功能的研究进展;窦剑明等;《生命科学》;20170228;第29卷(第2期);第124页左栏 2，第128页右栏第1段 * |
| Deleted in breast cancer-1 (DBC-1) in the interface between metabolism, aging and cancer;Eduardo Nunes CHINI 等;《Biosci. Rep.》;20131231;第33卷(第4期);第637页摘要，第641-642页结论 * |
| 乳腺癌缺失基因1对肝癌生物学行为的影响;顾勇等;《武警医学》;20121015(第10期);第871页"摘要" * |
| 窦剑明等.DBC1的蛋白质结构与生物学功能的研究进展.《生命科学》.2017,第29卷(第2期), * |
| 顾勇等.乳腺癌缺失基因1对肝癌生物学行为的影响.《武警医学》.2012,(第10期), * |
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