CN109096373A - Synthetic oligopeptide and application thereof - Google Patents
Synthetic oligopeptide and application thereof Download PDFInfo
- Publication number
- CN109096373A CN109096373A CN201811128109.8A CN201811128109A CN109096373A CN 109096373 A CN109096373 A CN 109096373A CN 201811128109 A CN201811128109 A CN 201811128109A CN 109096373 A CN109096373 A CN 109096373A
- Authority
- CN
- China
- Prior art keywords
- lung cancer
- oligopeptides
- cell
- transfer
- invasion
- 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.)
- Granted
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K7/00—Peptides having 5 to 20 amino acids in a fully defined sequence; Derivatives thereof
- C07K7/04—Linear peptides containing only normal peptide links
- C07K7/08—Linear peptides containing only normal peptide links having 12 to 20 amino acids
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
- A61P35/04—Antineoplastic agents specific for metastasis
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
Abstract
The invention discloses synthetic oligopeptides and application thereof.Lung cancer is one of global incidence and the highest malignant tumour of the death rate, its disease incidence occurs, development and transfer all refer to extremely complex polygenes and regulate and control exception procedure in China in rising and rejuvenation trend in recent years.Therefore, the cause of disease of lung cancer is studied, pathogenesis has important clinical meaning.Invasion and transfer are the main reason for malignant tumour lead to death.The drug of prevention and treatment lung cancer metastasis, treatment lung cancer can be made by proliferation, migration and the invasion of inhibition NGAL gene expression inhibition lung carcinoma cell in oligopeptides provided by the invention.
Description
Technical field
The invention belongs to chemical fields, are related to synthetic oligopeptide and application thereof.
Background technique
Lung cancer is one of global incidence and the highest malignant tumour of the death rate, its disease incidence is in China in rising in recent years
With rejuvenation trend, occur, development and transfer all refer to extremely complex polygenes regulation exception procedure.Therefore, study of lung
The cause of disease, the pathogenesis of cancer have important clinical meaning.
Invasion and transfer are the main reason for malignant tumour lead to death.
Summary of the invention
The present invention is intended to provide synthetic oligopeptide and application thereof.
Technical solution of the present invention is as follows:
A kind of oligopeptides, it is characterised in that: sequence is as shown in Sequence NO.2.
Above-mentioned oligopeptides is used as the purposes of NGAL gene expression inhibitor.
Above-mentioned oligopeptides is used to prepare the purposes of the drug of prevention and treatment lung cancer metastasis.
Above-mentioned oligopeptides is used to prepare the purposes of the drug for the treatment of lung cancer.
The utility model has the advantages that
Lung cancer is one of global incidence and the highest malignant tumour of the death rate, its disease incidence is in China in rising in recent years
With rejuvenation trend, occur, development and transfer all refer to extremely complex polygenes regulation exception procedure.Therefore, study of lung
The cause of disease, the pathogenesis of cancer have important clinical meaning.Invasion and transfer are the main originals that malignant tumour leads to death
Cause.Oligopeptides provided by the invention can be by inhibiting proliferation, migration and the invasion of NGAL gene expression inhibition lung carcinoma cell, can be with
The drug that prevention and treatment lung cancer metastasis is made, treats lung cancer.
Detailed description of the invention
Fig. 1 is the influence of NGAL gene expression in 1~5 pair of lung cancer A549 cell of oligopeptides；
Fig. 2 is the influence of 1~5 pair of lung cancer A549 cell of oligopeptides proliferation；
Fig. 3 is the influence of 1~5 pair of lung cancer A549 cell transfer ability of oligopeptides；
Fig. 4 is the influence of 1~5 pair of lung cancer A549 cell invasive ability of oligopeptides.
Specific embodiment
It is specific with reference to the accompanying drawings and examples to introduce essentiality content of the present invention, but guarantor of the invention is not limited with this
Protect range.
One, experimental material
Oligopeptides 1~5 entrusts the synthesis of outsourcing mechanism, and purity is not less than 95%, and sequence is as follows:
Oligopeptides 1:RCRVFYKPWVRHQMRGRYN (Sequence NO.1)；
Oligopeptides 2:HPWYRKWNYRVRWRGWFMR (Sequence NO.2)；
Oligopeptides 3:QDRKWSYKSSYFRKRGRSRT (Sequence NO.3)；
Oligopeptides 4:PHPEGEFRTKMEYRWEYRVR (Sequence NO.4)；
Oligopeptides 5:PHPEDEFATKFEYRWGYRVR (Sequence NO.5).
Fetal calf serum, DMEM high glucose medium are purchased from Gbico company；TRIzol reagent is purchased from Invitrogen company；Instead
Transcript reagent box and quantitative PCR kit are purchased from TAKARA company；Tanswell invades cell and is purchased from millpore company.
The RT-PCR primer commission outsourcing mechanism synthesis of NGAL, GAPDH.
Two, experimental method
1, cell culture
Lung cancer A549 cell sets 37 DEG C, 5%CO with the DMEM culture medium containing 10% fetal calf serum2Under the conditions of cultivate.
2, it is grouped and is administered
The lung cancer A549 cell of logarithmic growth phase, random grouping administration:
Control group: the DMEM culture medium culture for containing 10% fetal calf serum is used；
1 group of oligopeptides: adding 5 μM of oligopeptides 1 on the basis of the control group and cultivate；
2 groups of oligopeptides: adding 5 μM of oligopeptides 2 on the basis of the control group and cultivate；
3 groups of oligopeptides: adding 5 μM of oligopeptides 3 on the basis of the control group and cultivate；
4 groups of oligopeptides: adding 5 μM of oligopeptides 4 on the basis of the control group and cultivate；
5 groups of oligopeptides: adding 5 μM of oligopeptides 5 on the basis of the control group and cultivate.
3, influence of the oligopeptides to NGAL gene expression in lung cancer A549 cell
Lung cancer A549 cell is incubated at containing in 10% fetal calf serum DMEM culture medium, the cell of logarithmic growth phase disappears
Change, count, 2 × 105A/hole is laid on 24 orifice plates, after cell is adherent, is grouped and is changed to corresponding according to above-mentioned group technology
Culture medium culture, every group sets 3 parallel holes, continues to cultivate 48h, discard supernatant, cell is collected, by TRIzol kit specification
Cell total rna is extracted, measures RNA concentration respectively with ultraviolet specrophotometer, is determined according to absorbance OD260 and OD280 ratio
Purity.5 μ g cell total rnas are taken respectively, by the reverse transcription of reverse transcription reagent box specification at cDNA, using cDNA as template application.Instead
Answer condition are as follows: 94 DEG C of preheating 3min, then 94 DEG C of 30s, 52 DEG C of 30s, 72 DEG C of 50s, expand 29 circulations, 72 DEG C of extension 10min.
PCR product carries out 1% agarose gel electrophoresis, and the gray value of each electrophoretic band is analyzed by Image J image analysis software, with
The ratio of the gray value of the gray value and GAPDH internal reference band of purpose band indicates the relative expression quantity of target gene mRNA.
The upstream primer of NGAL is 5 '-GAAGACAAAGACCCGCAAAAG-3 ', and primer sequence is 5 '-downstream
CTGGCAACCTGGAACAAAAG-3′.The upstream primer sequence of internal reference GAPDH is 5 '-ACCACAGTCCATGCCATCAC-3 ',
Downstream primer is 5 '-TCCACCACCCTGTGCTGTA-3 '.
4, inhibited proliferation of the mtt assay detection oligopeptides to lung cancer A549 cell
Lung cancer A549 cell is incubated at containing in 10% fetal calf serum DMEM culture medium, the cell of logarithmic growth phase disappears
Change, count, 4 × 104A/hole is laid on 96 orifice plates, after cell is adherent, is grouped and is changed to corresponding according to above-mentioned group technology
Culture medium culture, every group sets 6 parallel holes, continues to cultivate 48h, discard supernatant, and serum free medium dissolution is added in each hole
MTT is incubated for 4h, to measure absorbance value after dmso solution at microplate reader 570nm.
5, cell migration ability detects
Lung cancer A549 cell is incubated at containing in 10% fetal calf serum DMEM culture medium, the cell of logarithmic growth phase disappears
To change, supernatant is abandoned in centrifugation, and cell is resuspended using the DMEM culture medium of 10% fetal calf serum containing volume fraction, single cell suspension is made,
According to above-mentioned grouping and medication in 37 DEG C, volume fraction 5%CO248h is cultivated in saturated humidity incubator.
After cultivating 48h, cell is collected in digestion, cell suspension is made with plasma-free DMEM medium, with 2.5 × 104A/hole
Density be inoculated in 6 orifice plates, when cell in monolayer adherence and close to 100% fusion when, using 10 μ l sterile pipette heads in 6 holes
Scratch in plate, with serum-free medium, is placed in 37 DEG C, 5%CO after cleaning suspension cast-off cells2In incubator, respectively at draw
0 and 48h takes pictures under inverted light microscope after trace, calculates its scratch healing rate, and experiment is repeated 3 times.Scratch healing rate=(0h
Scratch distance after scratch distance -48h)/0h scratch distance × 100%.
6, cell invasion ability detects
Lung cancer A549 cell is incubated at containing in 10% fetal calf serum DMEM culture medium, the cell of logarithmic growth phase disappears
To change, supernatant is abandoned in centrifugation, and cell is resuspended using the DMEM culture medium of 10% fetal calf serum containing volume fraction, single cell suspension is made,
According to above-mentioned grouping and medication in 37 DEG C, volume fraction 5%CO248h is cultivated in saturated humidity incubator.
After cultivating 48h, cell is collected in digestion, cell suspension is prepared with plasma-free DMEM medium, with 2.5 × 105/ hole
Density is laid on the cell Transwell upper layer chamber.Then, the DMEM culture medium of 20% fetal calf serum containing volume fraction is added
The cell Transwell lower chamber.After culture for 24 hours, the thin of non-invasion is removed from the cell Transwell upper layer chamber with cotton swab
Born of the same parents take the cell Transwell, with PBS washing 3 times, the fixed 30min of methanol, using 0.1% violet staining 30min, in inversion
Optical microphotograph sem observation counts.5 visuals field are randomly selected to count.
7, statistical analysis
Using 19.0 statistics software of SPSS, experiment is repeated 3 times, and data information is indicated with means standard deviation, sample standard deviation
Number is relatively examined using t between group.P < 0.05 is that difference has significant.
Three, experimental result
1, in 1~5 pair of lung cancer A549 cell of oligopeptides NGAL gene expression influence
The results are shown in Table 1 and Fig. 1, oligopeptides 1~4 can significantly inhibit the expression of NGAL gene in lung cancer A549 cell,
And oligopeptides 5 to the expression of NGAL gene in lung cancer A549 cell without obvious inhibiting effect.
The influence of NGAL gene expression in table 1 oligopeptides, 1~5 pair of lung cancer A549 cell
Neutrophil gelatinase-associated lipocalin (NGAL) is a member of lipocalin family.Research
It was found that NGAL high expression in cancerous lung tissue, it may be possible to treat one of lung cancer potential target spot (bibliography: lung squamous cancer and
The immunohistochemical study that NGAL is expressed in gland cancer, experiment and laboratory medicine, 01 phase in 2014).
2, the influence of 1~5 pair of lung cancer A549 cell of oligopeptides proliferation
It the results are shown in Table 2 and Fig. 2, oligopeptides 1~4 can significantly inhibit the proliferation of lung cancer A549 cell, and oligopeptides 5 is to lung cancer
The proliferation of A549 cell is without obvious inhibiting effect.
The influence of 2 1~5 pair of lung cancer A549 cell of oligopeptides of table proliferation
3, the influence of 1~5 pair of lung cancer A549 cell transfer ability of oligopeptides
It the results are shown in Table 3 and Fig. 3, oligopeptides 1~4 can significantly inhibit the transfer ability of lung cancer A549 cell, and oligopeptides 5 is to lung
The transfer ability of cancer A549 cell is without obvious inhibiting effect.
The influence of 3 1~5 pair of lung cancer A549 cell transfer ability of oligopeptides of table
4, the influence of 1~5 pair of lung cancer A549 cell invasive ability of oligopeptides
It the results are shown in Table 4 and Fig. 4, oligopeptides 1~4 can significantly inhibit the invasive ability of lung cancer A549 cell, and oligopeptides 5 is to lung
The invasive ability of cancer A549 cell is without obvious inhibiting effect.
The influence of 4 1~5 pair of lung cancer A549 cell invasive ability of oligopeptides of table
Lung cancer is one of global incidence and the highest malignant tumour of the death rate, its disease incidence is in China in rising in recent years
With rejuvenation trend, occur, development and transfer all refer to extremely complex polygenes regulation exception procedure.Therefore, study of lung
The cause of disease, the pathogenesis of cancer have important clinical meaning.Invasion and transfer are the main originals that malignant tumour leads to death
Cause.Oligopeptides provided by the invention can be by inhibiting proliferation, migration and the invasion of NGAL gene expression inhibition lung carcinoma cell, can be with
The drug that prevention and treatment lung cancer metastasis is made, treats lung cancer.
The effect of above-described embodiment is specifically to introduce essentiality content of the invention, but those skilled in the art should know
Protection scope of the present invention should not be confined to the specific embodiment by road.
Sequence table
<110>Huaian An Lai Biotechnology Co., Ltd
<120>synthetic oligopeptide and application thereof
<160> 5
<170> SIPOSequenceListing 1.0
<210> 1
<211> 19
<212> PRT
<213>artificial sequence (Artificial Sequence)
<400> 1
Arg Cys Arg Val Phe Tyr Lys Pro Trp Val Arg His Gln Met Arg Gly
1 5 10 15
Arg Tyr Asn
<210> 2
<211> 19
<212> PRT
<213>artificial sequence (Artificial Sequence)
<400> 2
His Pro Trp Tyr Arg Lys Trp Asn Tyr Arg Val Arg Trp Arg Gly Trp
1 5 10 15
Phe Met Arg
<210> 3
<211> 20
<212> PRT
<213>artificial sequence (Artificial Sequence)
<400> 3
Gln Asp Arg Lys Trp Ser Tyr Lys Ser Ser Tyr Phe Arg Lys Arg Gly
1 5 10 15
Arg Ser Arg Thr
20
<210> 4
<211> 20
<212> PRT
<213>artificial sequence (Artificial Sequence)
<400> 4
Pro His Pro Glu Gly Glu Phe Arg Thr Lys Met Glu Tyr Arg Trp Glu
1 5 10 15
Tyr Arg Val Arg
20
<210> 5
<211> 20
<212> PRT
<213>artificial sequence (Artificial Sequence)
<400> 5
Pro His Pro Glu Asp Glu Phe Ala Thr Lys Phe Glu Tyr Arg Trp Gly
1 5 10 15
Tyr Arg Val Arg
20
Claims (4)
1. a kind of oligopeptides, it is characterised in that: sequence is as shown in Sequence NO.2.
2. the purposes that oligopeptides described in claim 1 is used as NGAL gene expression inhibitor.
3. the purposes that oligopeptides described in claim 1 is used to prepare the drug of prevention and treatment lung cancer metastasis.
4. the purposes that oligopeptides described in claim 1 is used to prepare the drug for the treatment of lung cancer.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811128109.8A CN109096373B (en) | 2018-09-27 | 2018-09-27 | Synthetic oligopeptides and uses thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811128109.8A CN109096373B (en) | 2018-09-27 | 2018-09-27 | Synthetic oligopeptides and uses thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN109096373A true CN109096373A (en) | 2018-12-28 |
| CN109096373B CN109096373B (en) | 2022-06-21 |
Family
ID=64867125
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201811128109.8A Active CN109096373B (en) | 2018-09-27 | 2018-09-27 | Synthetic oligopeptides and uses thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN109096373B (en) |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1974768A (en) * | 2006-10-24 | 2007-06-06 | 汕头大学医学院 | Transcriptional control element of human lung carcinoma cell NGAL gene promoter region |
| CN101421622A (en) * | 2006-02-17 | 2009-04-29 | 儿童医学中心公司 | Free NGAL as a biomarker for cancer |
| WO2012042061A1 (en) * | 2010-10-01 | 2012-04-05 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Methods for predicting the progression and treating a chronic kidney disease in a patient |
| WO2015177175A2 (en) * | 2014-05-22 | 2015-11-26 | Pieris Ag | Novel specific-binding polypeptides and uses thereof |
-
2018
- 2018-09-27 CN CN201811128109.8A patent/CN109096373B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101421622A (en) * | 2006-02-17 | 2009-04-29 | 儿童医学中心公司 | Free NGAL as a biomarker for cancer |
| CN1974768A (en) * | 2006-10-24 | 2007-06-06 | 汕头大学医学院 | Transcriptional control element of human lung carcinoma cell NGAL gene promoter region |
| WO2012042061A1 (en) * | 2010-10-01 | 2012-04-05 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Methods for predicting the progression and treating a chronic kidney disease in a patient |
| WO2015177175A2 (en) * | 2014-05-22 | 2015-11-26 | Pieris Ag | Novel specific-binding polypeptides and uses thereof |
Non-Patent Citations (1)
| Title |
|---|
| 唐健等: "沉默NGAL基因对肺癌细胞迁移及侵袭的影响", 《中国肺癌杂志》 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN109096373B (en) | 2022-06-21 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Miskolczi et al. | Collagen abundance controls melanoma phenotypes through lineage-specific microenvironment sensing | |
| Fan et al. | Metformin exerts anticancer effects through the inhibition of the Sonic hedgehog signaling pathway in breast cancer Retraction in/10.3892/ijmm. 2023.5227 | |
| Zhao et al. | Hypoxia-induced cell stemness leads to drug resistance and poor prognosis in lung adenocarcinoma | |
| Ying et al. | Transcriptome analysis of phycocyanin inhibitory effects on SKOV-3 cell proliferation | |
| Gao et al. | Long non-coding RNA LINC00324 exerts pro-tumorigenic effects on liver cancer stem cells by up-regulating Fas ligand via PU box binding protein | |
| Zhao et al. | Long non-coding RNA SNHG20 promotes bladder cancer via activating the Wnt/β-catenin signalling pathway | |
| Xu et al. | Hyperglycemia promotes Snail-induced epithelial–mesenchymal transition of gastric cancer via activating ENO1 expression | |
| Su et al. | Sevoflurane inhibits proliferation, invasion, but enhances apoptosis of lung cancer cells by Wnt/β-catenin signaling via regulating lncRNA PCAT6/miR-326 axis | |
| Wu et al. | Population and single‑cell transcriptome analyses reveal diverse transcriptional changes associated with radioresistance in esophageal squamous cell carcinoma | |
| Sheta et al. | The polypeptide GALNT6 displays redundant functions upon suppression of its closest homolog GALNT3 in mediating aberrant O-glycosylation, associated with ovarian cancer progression | |
| Cheng et al. | Pristimerin suppressed breast cancer progression via miR-542-5p/DUB3 axis | |
| Zhang et al. | CMIP is oncogenic in human gastric cancer cells | |
| Xu et al. | Upregulation of NFKBIZ affects bladder cancer progression via the PTEN/PI3K/Akt signaling pathway | |
| Xing et al. | Effects of ulinastatin on proliferation and apoptosis of breast cancer cells by inhibiting the ERK signaling pathway | |
| Wang et al. | Curcumol synergizes with cisplatin in osteosarcoma by inhibiting M2-like polarization of tumor-associated macrophages | |
| Lu et al. | Anti-colon cancer effects of Spirulina polysaccharide and its mechanism based on 3D models | |
| CN109096373A (en) | Synthetic oligopeptide and application thereof | |
| CN109096374A (en) | A kind of synthetic oligopeptide inhibiting lung cancer metastasis | |
| CN109206485A (en) | A kind of oligopeptides for preventing and treating lung cancer metastasis | |
| CN108998495B (en) | A kind of high-throughput screening method of anti-breast cancer medicines | |
| CN109180789A (en) | A kind of oligopeptides and pharmacy application | |
| Gao et al. | SPRED2: a novel regulator of epithelial-mesenchymal transition and stemness in hepatocellular carcinoma cells | |
| CN114949226B (en) | Application of B7-H3 gene inhibitor in preparation of medicines for treating non-small cell lung cancer | |
| Harati et al. | Tumor-associated fibroblasts promote the proliferation and decrease the doxorubicin sensitivity of liposarcoma cells | |
| CN110317872A (en) | It is used to prepare the molecular marked compound and kit of detection, prognosis and Diagnosis of Breast cancer product |
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 | ||
| TA01 | Transfer of patent application right | ||
| TA01 | Transfer of patent application right |
Effective date of registration: 20220402 Address after: 611130 No. 48-24, Jinwu street, Liucheng, Wenjiang District, Chengdu, Sichuan Applicant after: Sichuan Zhongjian huapeptide Biotechnology Co.,Ltd. Address before: 223001 room 112, 191 Hanhou Avenue, qingjiangpu District, Huai'an City, Jiangsu Province Applicant before: HUAI'AN ANLAI BIOTECHNOLOGY Co.,Ltd. |
|
| TA01 | Transfer of patent application right | ||
| TA01 | Transfer of patent application right |
Effective date of registration: 20220531 Address after: Floors 1-4, building 22, No. 20, Xiushui Road, pharmaceutical high tech Zone, Taizhou, Jiangsu 225300 Applicant after: Jiangsu Hengrui Biomedical Technology Co.,Ltd. Address before: 611130 No. 48-24, Jinwu street, Liucheng, Wenjiang District, Chengdu, Sichuan Applicant before: Sichuan Zhongjian huapeptide Biotechnology Co.,Ltd. |
|
| GR01 | Patent grant | ||
| GR01 | Patent grant |