CN110170051A - Application of the KLF12 albumen in preparation treatment non-small cell lung cancer drug - Google Patents
Application of the KLF12 albumen in preparation treatment non-small cell lung cancer drug Download PDFInfo
- Publication number
- CN110170051A CN110170051A CN201910431631.1A CN201910431631A CN110170051A CN 110170051 A CN110170051 A CN 110170051A CN 201910431631 A CN201910431631 A CN 201910431631A CN 110170051 A CN110170051 A CN 110170051A
- Authority
- CN
- China
- Prior art keywords
- klf12
- lung cancer
- cell lung
- small cell
- albumen
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
Landscapes
- Health & Medical Sciences (AREA)
- Veterinary Medicine (AREA)
- Chemical & Material Sciences (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Life Sciences & Earth Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Epidemiology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Organic Chemistry (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The present invention provides a kind of KLF12 albumen and applies in the drug of preparation treatment non-small cell lung cancer, and the amino acid sequence of the KLF12 albumen is as shown in SEQ ID No:1.The drug of the treatment non-small cell lung cancer includes: by the double stranded RNA of RNA AF panel KLF12 gene expression, or the protein for inhibiting KLF12 protein active, or the small molecule compound for inhibiting KLF12 protein function.KLF12 albumen of the invention promotes the expression of PD-L1 in non-small cell lung cancer by transcription, striking low KLF12 by siRNA can cause the expression of PD-L1 to be lowered, plasmid transient expression KLF12 can significantly raise the transcriptional level of PD-L1, provide new therapeutic targets and effective new drug for prevention and treatment non-small cell lung cancer.
Description
Technical field
The invention belongs to pharmaceutical field, it is related to a kind of application of KLF12 albumen in preparation treatment non-small cell lung cancer drug.
Background technique
Programmed death receptor -1 (Programmed death-1, PD-1) and its ligand (Programmed death
Ligand-1, PD-L1) combine after formed inhibitive ability of immunity microenvironment, key effect, Duo Geji are played in tumor immune escape
Controlling for clinical blood system tumor and solid tumor has been successfully applied in the monoclonal antibody drug for blocking PD-1/PD-L1 to combine
It treats, it is the breakthrough in immunotherapy of tumors field that some patientss, which even can achieve " clinical cure " state,.In view of PD-
Key effect of the 1/PD-L1 in tumor immune escape, the medicament research and development for targeting PD-1/PD-L1 also become a big research hotspot.
The monoclonal antibody of existing 5 kinds of PD-1 or PD-L1 is successively ratified to list at present, is used for melanoma and Huo Qijin lymph earliest
The treatment of tumor is then also applied in the entity tumors such as non-small cell lung cancer, bladder cancer.Although PD-1/PD-L1 monoclonal
Antibody has remarkable effect for oncotherapy, but antibody drug also faces low single medicine response rate, adverse reaction rate height, drug partly
A series of problems, such as phase length, poor controllability of declining.Compared with macromolecular antibody drug, small-molecule drug usually has organ or tumour
It is good penetrability, small to immune system stimulation and the advantages that can take orally.Therefore, the small-molecule drug for being directed to PD-1/PD-L1 is found
It is considered as the important channel for overcoming antibody drug defect, it has also become the research hotspot in the field.JQ1 at present, eFT508 etc. are small
Molecular drug is in the preclinical study stage, and the exploitation of these micromolecular inhibitors is mainly based upon the expression regulation of PD-L1
What mechanism carried out, thus the regulatory mechanism for fully stating PD-L1 is significant to the research and development of small-molecule drug.
People's KLF12 albumen is from KLF12 gene (Genebank No.NM_007249.4) coding, is Kr ü pple sample
One of the factor (Kr ü pple-like factor, KLF) family member.The family protein is a kind of with homologous dna bond area
Transcription factor, containing 3 highly conserved classical cysteine/histidine (Cys2/His2) zinc fingers, which can be tied
It closes on " CACCC-box " and " GC-box " of DNA, plays transcriptional control effect.KLF12 is initially found to can be used as repressor
Inhibit -2 α of transcription factor AP-1, is mainly expressed in the tissue such as brain, kidney, liver, lung.Most researchs at present are thought that KLF12 is played and are promoted
Function of tumor.In colorectal cancer, KLF12 can promote tumour growth by direct activation early growth reactive protein EGR1,
The level of KLF12 and EGR1 is jointly related to the poor prognosis of tumour, can be used as potential diagnostic marker and therapy target.
MiR-382 can inhibit the growth of osteosarcoma cell by targeting KLF12, inhibit miR-382 or overexpression KLF12 that can promote
The growth of osteosarcoma.Although having the relevant report of some KLF12 and tumour, KLF12 is to the tumour immunities such as PD-L1 correlation
Albumen is still unknown with the presence or absence of regulation relationship.
Summary of the invention
The object of the present invention is to provide a kind of application of KLF12 albumen in preparation treatment non-small cell lung cancer drug, especially
Its application caused by preparation is increased because of the protein level of tumour cell PD-L1 in non-small cell lung cancer drug.The KLF12 egg
White amino acid sequence is as shown in SEQ ID No:1.The drug of the treatment non-small cell lung cancer includes: to interfere to press down by RNA
The double stranded RNA of KLF12 gene expression processed, or the protein for inhibiting KLF12 protein active, or for inhibiting KLF12
The small molecule compound of protein function.
People's KLF12 albumen is from KLF12 gene (Genebank No.NM_007249.4) coding, is Kr ü pple sample
One of factor family member.People's KLF12 albumen is the transcription factor with homologous dna bond area, highly conserved containing 3
Classical cysteine/histidine (Cys2/His2) zinc fingers, the structure can be coupled to " CACCC-box " and " GC- of DNA
On box ", transcriptional control effect is played.Most researchs at present think KLF12 played in kinds of tumors promotion tumor development and
The effect of clinical poor prognosis, but KLF12 is still unknown with the presence or absence of regulation relationship to the tumour immunities GAP-associated protein GAP such as PD-L1
's.
Present invention experiment is expressed non-small with low expression by using sorting flow cytometry sorting PD-L1 film surface height
Cell lung cancer cell NCI-H460 to amplification cultivation and verifies this two groups of cell sample presentations progress full genome cores after PD-L1 expression
Higher expression is presented in piece analysis, discovery KLF12 in the highly expressed cell of PD-L1, and the two has apparent be positively correlated.
Using siRNA technology in the higher non-small cell lung cancer cell NCI-H460 and NCI-H1299 of two plants of KLF12 protein expression levels
Middle silencing KLF12 can significantly lower the transcriptional level of PD-L1, and in the lower non-small cell lung cancer of KLF12 protein expression level
Plasmid transient expression KLF12 can significantly raise the transcriptional level of PD-L1 in cell A549, and prompting KLF12 may be regulation PD-L1
Transcription factor, have no document report, be worth further investigation.
The present invention is experiments have shown that KLF12 is struck by the expression of PD-L1 in transcription promotion non-small cell lung cancer by siRNA
Low KLF12 can cause the expression of PD-L1 to be lowered, and plasmid transient expression KLF12 can promote the expression of PD-L1.Non-small cell
Lung cancer patient sample display KLF12 and PD-L1 expression has positive correlation, and after KLF12 expression and lung cancer patient long term survival
Poor prognosis there are certain correlations.Therefore, the present invention not only have found PD-L1 is completely new in non-small cell lung cancer regulation because
Son, and prompt to pass through the double stranded RNA of RNA AF panel KLF12 gene expression, or for inhibiting KLF12 protein active
Protein, or for inhibiting the small molecule compound of KLF12 protein function to can be used as the potential plan of Treatment for Non-small Cell Lung
Slightly, KLF12 also is likely used for the indication and patient's prognosis evaluation of PD-L1 expression.KLF12 albumen of the present invention is prevention and treatment non-small cell
Lung cancer provides new therapy target and effective new drug.
Detailed description of the invention
Fig. 1 is the CD274 and KLF12 that the present invention detects PD-L1 relatively high expression and low expression cell by q-RT-PCR
The transcriptional level of equal transcriptional controls related protein encoding gene.
Fig. 2 is that the present invention detects the influence for striking low KLF12 to PD-L1 transcription and protein expression by siRNA technology.Wherein
A figure be in non-small cell lung cancer NCI-H1299 cell respectively with siRNA technology silencing KLF12 and PD-L1 to KLF12 and
The influence of CD274mRNA level；B figure is influence of the silencing KLF12 48h to KLF12 and PD-L1 protein level；C figure is to pass through
Quantitative statistics figure of the gray analysis to B figure.D, E, F figure are the reproducible results on non-small cell lung cancer NCI-H460 cell.
Fig. 3 is influence of the present invention by plasmid transient expression KLF12 to PD-L1 transcription and protein expression.Wherein A figure is
Plasmid is overexpressed the KLF12 influence to KLF12 and PD-L1 protein level for 24 hours in Non-small Cell Lung Cancer A 549；B figure is
By gray analysis to the quantitative statistics figure of A figure；C, D figure is the reproducible results on non-small cell lung cancer NCI-H1299 cell.
Fig. 4 is that the present invention passes through Immunohistochemical Method detection KLF12 and PD-L1 table in non-small cell lung cancer patient's tumor tissue
The correlation reached.
Fig. 5 is the present invention by investigating TCGA database investigation KLF12 and Patients with Non-small-cell Lung by UCSC × ena
The correlation of clinical prognosis.
Specific embodiment
The present invention is further described in conjunction with the accompanying drawings and embodiments.
Embodiment 1
The relatively high expression (NCI-H460-PD-L1 of PD-L1 is detected using q-RT-PCRH) and low expression (NCI-H460-PD-L1L)
The transcriptional level of the transcriptional controls related protein encoding gene such as CD274 and KLF12 of cell.Specific step is as follows:
Choose the non-small cell lung cancer NCI-H460-PD- that same batch of secondary culture of genetic chip sample presentation is carried out after airflow classification
L1L and NCI-H460-PD-L1H cell inoculation is in 6 orifice plates, 1.0 × 106Cells/well, 37 DEG C, 5%CO2It is cultivated in incubator
Overnight.Its mRNA is collected, the transcriptional controls GAP-associated protein GAPs such as two kinds of cells CD274 and KLF12 are detected by q-RT-PCR and encode base
The transcriptional level of cause.Specifically the transcriptional controls GAP-associated protein GAP such as CD274 and KLF12 is compiled in non-small cell lung cancer NCI-H460 cell
The transcriptional level of code gene is shown in Fig. 1.
Embodiment 2
SiRNA technology is used to strike in the higher non-small cell lung cancer NCI-H1299 of KLF12 expression and NCI-H460 cell low
KLF12 is transcribed to PD-L1 and the influence of protein expression.Specific step is as follows:
Relatively high non-small cell lung cancer NCI-H1299 and the NCI-H460 cell of KLF12 protein expression level is chosen to connect respectively
Kind is in 6 orifice plates, and 1.0 × 106Cells/well, 37 DEG C, 5%CO2Overnight incubation in incubator；Next day uses siKLF12 (#1 and #
2) low KLF12 is struck, culture was changed liquid after 6 hours, and continue to cultivate 24 hours collection mRNA, cultivates 48 hours collection total proteins,
The influence for striking low KLF12 to PD-L1 transcription and protein expression is investigated by q-RT-PCR and western blot respectively.Specifically
SiKLF12 (#1 and #2) non-small cell lung cancer NCI-H1299 and NCI-H460 cell is transcribed to PD-L1 and the influence of protein expression
See Fig. 2.
Embodiment 3
It is expressed in KLF12 and transfects pcDNA3.0-KLF12-HA plasmid in lower A549 cell and higher NCI-H1299 cell
With the influence of the corresponding unloaded protein expression level to PD-L1.Specific step is as follows:
Non-small Cell Lung Cancer A 549 and NCI-H1299 cell inoculation are chosen in 6 orifice plates, 1 × 106Cells/well, 37 DEG C,
5%CO2Overnight incubation in incubator.Next day plasmid transient expression KLF12, culture were changed liquid after 6 hours, and it is small to continue culture 24
When collect total protein, investigated by western blot and be overexpressed influence of the KLF12 to PD-L1 protein expression.Specifically non-small
Fig. 3 is shown in influence of cell lung cancer A549 and NCI-H460 the bioblast transient expression KLF12 to PD-L1 protein expression.
Embodiment 4
Using Immunohistochemical Method investigate KLF12 and PD-L1 expression there are correlations in non-small cell lung cancer patient's tumor tissue.
Specific step is as follows:
33 non-small cell lung cancer patient's tumor tissues samples are soaked in 10% neutral formalin (4% formaldehyde), and to fix 48 small
When more than, tissue is put into dewaterer and is dehydrated, using paraffin embedding, paraffin section is carried out, obtains the slice with a thickness of 3 μm.
Then the expression of KLF12 and PD-L1 albumen is analyzed by immunohistochemical staining.Fig. 4 is shown in specific effect.
Embodiment 5
The correlation of KLF12 and Patients with Non-small-cell Lung clinical prognosis are investigated using UCSC × ena investigation TCGA database.Tool
Steps are as follows for body:
TCGA database has been investigated by UCSC × ena, has depicted KLF12 and adenocarcinoma of lung (LUAD), lung using bi-component group method
Gland cancer (LUSC) and both summation patient (LUNG) overall survival (OS) Kaplan Meier Plot with analyze KLF12 with
The correlation of Patients with Non-small-cell Lung clinical prognosis.Particularly relevant property is shown in Fig. 4.
Sequence table
<110>Zhejiang University
<120>application of the KLF12 albumen in preparation treatment non-small cell lung cancer drug
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 402
<212> PRT
<213>KLF12 albumen (Kr ü pple-like factor 12)
<400> 1
Met Asn Ile His Met Lys Arg Lys Thr Ile Lys Asn Ile Asn Thr Phe
1 5 10 15
Glu Asn Arg Met Leu Met Leu Asp Gly Met Pro Ala Val Arg Val Lys
20 25 30
Thr Glu Leu Leu Glu Ser Glu Gln Gly Ser Pro Asn Val His Asn Tyr
35 40 45
Pro Asp Met Glu Ala Val Pro Leu Leu Leu Asn Asn Val Lys Gly Glu
50 55 60
Pro Pro Glu Asp Ser Leu Ser Val Asp His Phe Gln Thr Gln Thr Glu
65 70 75 80
Pro Val Asp Leu Ser Ile Asn Lys Ala Arg Thr Ser Pro Thr Ala Val
85 90 95
Ser Ser Ser Pro Val Ser Met Thr Ala Ser Ala Ser Ser Pro Ser Ser
100 105 110
Thr Ser Thr Ser Ser Ser Ser Ser Ser Arg Leu Ala Ser Ser Pro Thr
115 120 125
Val Ile Thr Ser Val Ser Ser Ala Ser Ser Ser Ser Thr Val Leu Thr
130 135 140
Pro Gly Pro Leu Val Ala Ser Ala Ser Gly Val Gly Gly Gln Gln Phe
145 150 155 160
Leu His Ile Ile His Pro Val Pro Pro Ser Ser Pro Met Asn Leu Gln
165 170 175
Ser Asn Lys Leu Ser His Val His Arg Ile Pro Val Val Val Gln Ser
180 185 190
Val Pro Val Val Tyr Thr Ala Val Arg Ser Pro Gly Asn Val Asn Asn
195 200 205
Thr Ile Val Val Pro Leu Leu Glu Asp Gly Arg Gly His Gly Lys Ala
210 215 220
Gln Met Asp Pro Arg Gly Leu Ser Pro Arg Gln Ser Lys Ser Asp Ser
225 230 235 240
Asp Asp Asp Asp Leu Pro Asn Val Thr Leu Asp Ser Val Asn Glu Thr
245 250 255
Gly Ser Thr Ala Leu Ser Ile Ala Arg Ala Val Gln Glu Val His Pro
260 265 270
Ser Pro Val Ser Arg Val Arg Gly Asn Arg Met Asn Asn Gln Lys Phe
275 280 285
Pro Cys Ser Ile Ser Pro Phe Ser Ile Glu Ser Thr Arg Arg Gln Arg
290 295 300
Arg Ser Glu Ser Pro Asp Ser Arg Lys Arg Arg Ile His Arg Cys Asp
305 310 315 320
Phe Glu Gly Cys Asn Lys Val Tyr Thr Lys Ser Ser His Leu Lys Ala
325 330 335
His Arg Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Thr Trp Glu
340 345 350
Gly Cys Thr Trp Lys Phe Ala Arg Ser Asp Glu Leu Thr Arg His Tyr
355 360 365
Arg Lys His Thr Gly Val Lys Pro Phe Lys Cys Ala Asp Cys Asp Arg
370 375 380
Ser Phe Ser Arg Ser Asp His Leu Ala Leu His Arg Arg Arg His Met
385 390 395 400
Leu Val
Claims (3)
1. a kind of application of KLF12 albumen in preparation treatment non-small cell lung cancer drug, the amino acid sequence of the KLF12 albumen is such as
Shown in SEQ ID No:1.
2. application according to claim 1, which is characterized in that egg of the non-small cell lung cancer because of tumour cell PD-L1
White level, which increases, to be caused.
3. application according to claim 1, which is characterized in that the drug of the treatment non-small cell lung cancer includes: logical
The double stranded RNA of RNA AF panel KLF12 gene expression, or the protein for inhibiting KLF12 protein active are crossed, or is used
In the small molecule compound for inhibiting KLF12 protein function.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910431631.1A CN110170051A (en) | 2019-05-23 | 2019-05-23 | Application of the KLF12 albumen in preparation treatment non-small cell lung cancer drug |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910431631.1A CN110170051A (en) | 2019-05-23 | 2019-05-23 | Application of the KLF12 albumen in preparation treatment non-small cell lung cancer drug |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN110170051A true CN110170051A (en) | 2019-08-27 |
Family
ID=67691883
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201910431631.1A Pending CN110170051A (en) | 2019-05-23 | 2019-05-23 | Application of the KLF12 albumen in preparation treatment non-small cell lung cancer drug |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN110170051A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111454335A (en) * | 2020-04-28 | 2020-07-28 | 上海市农业科学院 | Protein AMU3 and application thereof in preparation of anti-lung cancer drugs |
| CN112980799A (en) * | 2021-02-10 | 2021-06-18 | 南京鼓楼医院 | Method for constructing KLF12 high-expression mouse and application of method in construction of folate-independent neural tube defect mouse model |
-
2019
- 2019-05-23 CN CN201910431631.1A patent/CN110170051A/en active Pending
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111454335A (en) * | 2020-04-28 | 2020-07-28 | 上海市农业科学院 | Protein AMU3 and application thereof in preparation of anti-lung cancer drugs |
| CN111454335B (en) * | 2020-04-28 | 2022-09-23 | 上海市农业科学院 | Protein AMU3 and application thereof in preparation of anti-lung cancer drugs |
| CN112980799A (en) * | 2021-02-10 | 2021-06-18 | 南京鼓楼医院 | Method for constructing KLF12 high-expression mouse and application of method in construction of folate-independent neural tube defect mouse model |
| CN112980799B (en) * | 2021-02-10 | 2022-12-27 | 南京鼓楼医院 | Method for constructing KLF12 high-expression mouse and application of method in construction of folate-independent neural tube defect mouse model |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Jing et al. | BRD4 inhibition suppresses PD-L1 expression in triple-negative breast cancer | |
| CN112522394B (en) | Novel exosome release related target and application thereof in tumor monitoring and inhibition | |
| CN109420170A (en) | Novel tumor microenvironment related target TAK1 and its application in inhibition tumour | |
| CN111073979A (en) | Gastric cancer treatment method for blocking CCL28 chemotactic pathway | |
| CN107299133A (en) | A kind of mark SPOP of prostate cancer and its application in oncotherapy is instructed | |
| CN110170051A (en) | Application of the KLF12 albumen in preparation treatment non-small cell lung cancer drug | |
| Ge et al. | Circular RNA CELF1 drives immunosuppression and anti-PD1 therapy resistance in non-small cell lung cancer via the miR-491-5p/EGFR axis | |
| CN111040032B (en) | Application of bidirectional regulator in preparation of cell aging and tumor diagnosis or regulation preparation | |
| KR101596166B1 (en) | A Use of microRNA for Ddiagnosing and Treating Brest Cancer | |
| CN101273131A (en) | Pancreatic cancer related gene CST6 and GABRP | |
| CN116287275B (en) | Use of PTGR1 as a CDK4/6 inhibitor and metformin combination guide marker | |
| Feng et al. | Effect of solanum lyratum polysaccharide on malignant behaviors of lung cancer cells by regulating the circ_UHRF1/miR-513b-5p axis | |
| CN105214077B (en) | Application of the USP33 in tumour | |
| CN105189786A (en) | FALZ for use as a target for therapies to treat cancer | |
| CN107488735B (en) | MiR-339-5p is inhibiting the application in prostate cancer with osseous metastasis and TGF-β signal path | |
| CN111996251B (en) | Application of malignant glioma biomarker | |
| CN113564175A (en) | Gene, protein, vector, lentivirus with hexokinase 2 inhibitory activity and application | |
| CN106554962B (en) | Prevention, diagnosis and treatment of GPR160 overexpressing cancers | |
| CN113413460A (en) | Application of TRAF6 inhibitor in preparation of melanoma drugs | |
| CN108277276B (en) | Application of long-chain non-coding RNA and composition thereof in diagnosis/treatment of gastric cancer | |
| CN115747337A (en) | Application of CCT2 as glioma marker | |
| CN107308450A (en) | Glycerokinase as the therapeutic targets of carbohydrate metabolism disturbance disease purposes | |
| Ji | Effect of lncRNA LINC00483 on Taxol Resistance of Hepatocellular Carcinoma Cells by Targeting miR-646 | |
| CN105671195A (en) | Application of miR-520c nucleotide, pharmaceutical composition and kit | |
| CN118010995A (en) | PHB2 as tumor immunotherapy target and application thereof in drug development |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20190827 |
|
| WD01 | Invention patent application deemed withdrawn after publication |