CN108018354A - New applications of the MicroRNA-34 in prostate cancer transfer treatment is suppressed - Google Patents
New applications of the MicroRNA-34 in prostate cancer transfer treatment is suppressed Download PDFInfo
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- CN108018354A CN108018354A CN201711370384.6A CN201711370384A CN108018354A CN 108018354 A CN108018354 A CN 108018354A CN 201711370384 A CN201711370384 A CN 201711370384A CN 108018354 A CN108018354 A CN 108018354A
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Abstract
The invention discloses miR 34b and miR 34c to suppress metastatic prostate cancer cell migration and invasion and attack.New biological markers are provided for treatment metastatic prostate cancer.
Description
Technical field
The present invention relates to the correlation technique of cell biology, genomics and field of bioinformatics.
Background technology
MicroRNA（miRNA）As important gene expression regulation molecule, cancer can be played in tumour generating process
The effect of gene or tumor suppressor gene.It is miR-34a, miR-34b and miR-34c respectively that, which there are three members in miR-34 families,.They
Expression in the prostate gland cancer cell of Metastatic potential has notable difference, and wherein miR-34b and miR-34c differences are shown
Write.
The content of the invention
The purpose of the present invention is：It is found that miR-34b and miR-34c has and suppresses prostate cancer transferance, is forefront
Gland cancer transfer treatment provides new biomarker.
The technical scheme is that：MicroRNA-34 family members miR-34b is thin as metastatic prostate cancer is suppressed
Born of the same parents migrate and invasion and attack, suppress the application of cancer metastasis.
MicroRNA-34 family members miR-34c suppresses cancer as metastatic prostate cancer cell migration and invasion and attack are suppressed
The application of disease transfer,.
MicroRNA-34 family members miR-34b and miR-34c are used in combination as suppression metastatic prostate cancer cell
Migration and invasion and attack, suppress the application of cancer metastasis.
One of MicroRNA-34 family members miR-34b and miR-34c are used in combination as treatment metastatic forefront
Gland cancer provides biological markers.
The present invention process be
1. it is thin in high metastatic potential prostate gland cancer cell PC-3 and low metastatic potential prostate cancer to detect miR-34b and miR-34c
Expression quantity in born of the same parents' DU145 cells.
2. transfect mimic and inhibitor high expression and suppress miR-34b and miR-34c in prostate gland cancer cell.
3. utilize the high expression of Transwell experiment detections and the prostate gland cancer cell of low expression miR-34b and miR-34c
Migration and invasive ability.
The beneficial effects of the invention are as follows：MiR-34b and miR-34c suppresses metastatic prostate cancer cell migration and invasion and attack.
New biological markers are provided for treatment metastatic prostate cancer.
Brief description of the drawings
Attached drawing 1 detects tables of the miR-34b and miR-34c in prostate gland cancer cell PC-3, DU145 for real time fluorescent quantitative
Up to amount.
Migration and invasive ability of the attached drawing 2 for transfection miR-34b/c mimic and NC and the PC-3 cells of untransfected.
Attached drawing 3 is the statistics of the cell quantity of migration and invasion and attack in Fig. 2.
Migration and invasion and attack energy of the attached drawing 4 for transfection miR-34b/c inhibitor and NC and the DU145 cells of untransfected
Power.
The statistics for the cell quantity for migrating and attacking in 5 Fig. 4 of attached drawing.
Embodiment
The embodiment of the present invention：
Transwell methods, which detect miR-34b and miR-34c, influences migration of prostate cancer cells and invasive ability
Cell and other reagents
Human prostate cancer cell line PC-3, DU145 come from Toronto Sunnybrook research centers；DMEM culture mediums
And pancreatin is purchased from Hyclone companies of the U.S.；Hyclone（FBS）It is purchased from Zhejiang Tian Hang bio tech ltd；TRIzol®
reagent 、Lipofectamine®3000 reagent are purchased from Invitrogen companies of the U.S.；Primer, mimic and
Inhibitor is purchased from GuangZhou, China Rui Bo bio tech ltd；CDNA Reverse Transcriptase kits, UItraSYBR Green
It is ShiJi Co., Ltd that qPCR Mixture, which are purchased from health,；Opti-MEM culture mediums are purchased from Gibco companies of the U.S.；Transwell is purchased from
Corning Incorporated；Matrigel is purchased from BD Biosciences companies of the U.S..
Cell migration detects
1. miR-34b/c detection of expression
1.1 culture PC-3 and DU145 cells, collect cell and extract total serum IgE with TRIzol methods, use cDNA Reverse Transcriptase kits
It is cDNA by RNA reverse transcriptions, is detected for PCR.
1.2 real time fluorescent quantitatives detect expression quantity of the miR-34b/c in PC-3 and DU145 cells.
2. transfect miR-34b/c mimic, inhibitor and control Negative Control（NC）To prostate cancer
Cell
Use Lipofectamine®3000 reagent transfect miR-34b/c mimic and NC to PC-3 cells；miR-
34b/c inhibitor and NC are transfected into DU145 cells.By Lipofectamine during transfection®3000 and mimic（50nM）
And inhibitor（100nM）It is mixed into Opti-MEM culture mediums and forms transfection composite, adds to culture medium containing Opti-MEM
Cell in transfect 48h.Cell extraction total serum IgE is collected, reverse transcription carries out real-time fluorescence quantitative PCR detection transfection effect into cDNA
Rate.
3. Transwell methods detect cell migration and invasive ability
Three groups of the cell of collection transfection mimic/inhibitor and NC48h cells and untransfected, digestion and suspension cell, every group
Take 1 × 105/ ml cells are diluted in 200 μ l Opti-MEM culture medium inoculateds in Transwell plates per in the upper chamber of hole, add in lower room
Enter the 700 μ l culture mediums of DMEM containing 10%FBS, put in 37 DEG C of incubators and cultivate 14h.Transwell cells are taken out with 75% ethanol to be consolidated
Determine cell, the cell number migrated after 0.1% violet staining with inverted phase contrast microscope observation to lower room determines its transfer ability.
Detection cell invasion ability need to spread upper interior into 200 μ l 1 before inoculating cell:5 diluted Matrigel.
4. the results show：
MiR-34b/c low expressions in high metastatic prostate cancer cell PC-3, in low metastatic prostate cancer cell DU145
High expression in cell.The PC-3 cells of transfection miR-34b/c mimic are than transfection NC controls and the cell migration of untransfected and invade
It is weak to attack power；The DU145 cells of transfection miR-34b/c inhibitor are than transfection NC controls and the cell migration of untransfected and invade
It is strong to attack power.Therefore miR-34b/c has the function that to suppress migration of prostate cancer cells and invasion and attack.
Claims (4)
1.MicroRNA-34 family member miR-34b suppresses cancer as metastatic prostate cancer cell migration and invasion and attack are suppressed
The application of transfer.
2.MicroRNA-34 family member miR-34c suppresses cancer as metastatic prostate cancer cell migration and invasion and attack are suppressed
The application of transfer,.
Moved 3.MicroRNA-34 family member miR-34b and miR-34c are used in combination as suppression metastatic prostate cancer cell
Move and attack, suppress the application of cancer metastasis.
4.MicroRNA-34 one of family member miR-34b and miR-34c are used in combination to treat metastatic prostate
Cancer provides biological markers.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711370384.6A CN108018354A (en) | 2017-12-19 | 2017-12-19 | New applications of the MicroRNA-34 in prostate cancer transfer treatment is suppressed |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
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| CN201711370384.6A CN108018354A (en) | 2017-12-19 | 2017-12-19 | New applications of the MicroRNA-34 in prostate cancer transfer treatment is suppressed |
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| Publication Number | Publication Date |
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| CN108018354A true CN108018354A (en) | 2018-05-11 |
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| CN201711370384.6A Withdrawn CN108018354A (en) | 2017-12-19 | 2017-12-19 | New applications of the MicroRNA-34 in prostate cancer transfer treatment is suppressed |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110484605A (en) * | 2019-09-23 | 2019-11-22 | 中国人民解放军陆军军医大学第一附属医院 | A kind of gene and its preparation method and application of living cells in situ detection microRNA-34 |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101801419A (en) * | 2007-06-08 | 2010-08-11 | 米尔纳疗法公司 | Gene and path as the miR-34 regulation and control for the treatment of the target of intervening |
| CN102027129A (en) * | 2008-02-28 | 2011-04-20 | 俄亥俄州立大学研究基金会 | Microrna-based methods and compositions for the diagnosis, pronosis and treatment of prostate related disorders |
| CN105473742A (en) * | 2013-06-24 | 2016-04-06 | 米尔纳疗法公司 | Biomarkers of miR-34 activity |
| US20160362689A1 (en) * | 2012-08-29 | 2016-12-15 | City Of Hope | Differentially expressed microrna molecules for the treatment and diagnosis of cancer |
-
2017
- 2017-12-19 CN CN201711370384.6A patent/CN108018354A/en not_active Withdrawn
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101801419A (en) * | 2007-06-08 | 2010-08-11 | 米尔纳疗法公司 | Gene and path as the miR-34 regulation and control for the treatment of the target of intervening |
| CN102027129A (en) * | 2008-02-28 | 2011-04-20 | 俄亥俄州立大学研究基金会 | Microrna-based methods and compositions for the diagnosis, pronosis and treatment of prostate related disorders |
| US20160362689A1 (en) * | 2012-08-29 | 2016-12-15 | City Of Hope | Differentially expressed microrna molecules for the treatment and diagnosis of cancer |
| CN105473742A (en) * | 2013-06-24 | 2016-04-06 | 米尔纳疗法公司 | Biomarkers of miR-34 activity |
Non-Patent Citations (3)
| Title |
|---|
| BEATRIZ A WALTER等: "Comprehensive microRNA Profiling of Prostate Cancer", 《JOURNAL OF CANCER》 * |
| SHAHANA MAJID等: "miRNA-34b Inhibits Prostate Cancer through Demethylation, Active Chromatin Modifications, and AKT Pathways", 《CLIN CANCER RES》 * |
| 刘浩等: "miRNA一34e对人前列腺癌PC3细胞增殖和侵袭能力的影响", 《中国医师杂志》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110484605A (en) * | 2019-09-23 | 2019-11-22 | 中国人民解放军陆军军医大学第一附属医院 | A kind of gene and its preparation method and application of living cells in situ detection microRNA-34 |
| CN110484605B (en) * | 2019-09-23 | 2020-07-28 | 中国人民解放军陆军军医大学第一附属医院 | Gene for in-situ detection of microRNA-34 by living cells and preparation method and application thereof |
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