miR-637抑制结肠癌HCT116细胞生长和迁移的机制研究
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摘要
目的:分析miR-637对结肠癌HCT116细胞生长和迁移的影响。方法:分别构建miR-637过表达或者低表达的结肠癌HCT116细胞系。通过MTT方法检测miR-637对细胞增殖的作用,PI(propidium iodide)染色检测miR-637对细胞周期的影响,Annexin V-FITC/PI染色检测miR-637对细胞凋亡的作用。Western blot检测miR-637对细胞中CDK4和Bcl-2的影响。Transwell实验检测miR-637对细胞迁移的作用,进一步通过Western blot检测miR-637对细胞中MMP2的影响。结果:miR-637抑制HCT116细胞的增殖,抑制G_1/S期进程,促进细胞凋亡,Western blot结果显示,miR-637抑制CDK4和Bcl-2在HCT116细胞中的表达。Transwell实验指出,miR-637抑制细胞的迁移,Western blot结果指出miR-637抑制MMP2在HCT116细胞中的表达。结论:miR-637可以通过抑制CDK4、Bcl-2和MMP2的表达抑制HCT116细胞的生长和迁移。
Objective:To analyze the effect of miR-637 on the growth and migration of HCT116 cells.Methods:The HCT116 cell lines with overexpressed or low expression of miR-637 were constructed respectively.MTT was used to detect the effect of miR-637 on cell proliferation.PI(propidium iodide) staining was used to detect the effect of miR-637 on cell cycle.Annexin V-FITC/PI staining was used to detect the effect of miR-637 on apoptosis.The effects of miR-637 on CDK4 and Bcl-2 in cells were detected by Western blot.The effect of miR-637 on cell migration was detected by Transwell,and the effect of miR-637 on the MMP2 in cells was detected by Western blot.Results:miR-637 can inhibit the proliferation of HCT116 cells,inhibit G_1/S phase progression,and promote cell apoptosis.Western blot results showed that miR-637 inhibited CDK4 and Bcl-2 expression in HCT116 cells.Transwell experiments showed that miR-637 inhibited cell migration.Western blot results showed that miR-637 inhibited MMP2 expression in HCT116 cells.Conclusion:miR-637 inhibited the growth and migration of HCT116 cells by inhibiting the expression of CDK4,Bcl-2 and MMP2.
Objective:To analyze the effect of miR-637 on the growth and migration of HCT116 cells.Methods:The HCT116 cell lines with overexpressed or low expression of miR-637 were constructed respectively.MTT was used to detect the effect of miR-637 on cell proliferation.PI(propidium iodide) staining was used to detect the effect of miR-637 on cell cycle.Annexin V-FITC/PI staining was used to detect the effect of miR-637 on apoptosis.The effects of miR-637 on CDK4 and Bcl-2 in cells were detected by Western blot.The effect of miR-637 on cell migration was detected by Transwell,and the effect of miR-637 on the MMP2 in cells was detected by Western blot.Results:miR-637 can inhibit the proliferation of HCT116 cells,inhibit G_1/S phase progression,and promote cell apoptosis.Western blot results showed that miR-637 inhibited CDK4 and Bcl-2 expression in HCT116 cells.Transwell experiments showed that miR-637 inhibited cell migration.Western blot results showed that miR-637 inhibited MMP2 expression in HCT116 cells.Conclusion:miR-637 inhibited the growth and migration of HCT116 cells by inhibiting the expression of CDK4,Bcl-2 and MMP2.
引文
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[3] Xu XW,Zheng BA,Hu ZM,et al.Circular RNA hsa_circ_000984 promotes colon cancer growth and metastasis by sponging miR-106b [J].Oncotarget,2017,8(53):91674-91683.
[4] Stachowiak M,Flisikowskal T,Bauersachs S,et al.Altered microRNA profiles during early colon adenoma progression in a porcine model of familial adenomatous polyposis [J].Oncotarget,2017,8(56):96154-96160.
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[6] Pashaei E,Pashaei E,Ahmady M,et al.Meta-analysis of miRNA expression profiles for prostate cancer recurrence following radical prostatectomy [J].PLoS One,2017,12(6):e0179543.
[7] Kim Y,Noren Hooten N.Posttranscriptional regulation of the inflammatory marker C-reactive protein by the RNA-binding protein HuR and microRNA637[J].Mol Cell Biol,2015,35(24):4212-4221.
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[9] Li J,Li B,Ren C,et al.The clinical significance of circulating GPC1 positive exosomes and its regulative miRNAs in colon cancer patients [J].Oncotarget,2017,8(60):101189-101202.
[10] Hua FF,Liu SS,Zhu LH,et al.MiRNA-338-3p regulates cervical cancer cells proliferation by targeting MACC1 through MAPK signaling pathway[J].Eur Rev Med Pharmacol Sci,2017,21(23):5342-5352.
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[13] Zhang JF,He ML,Fu WM,et al.Primate-specific microRNA-637 inhibits tumorigenesis in hepatocellular carcinoma by disrupting signal transducer and activator of transcription 3 signaling [J].Hepatology,2011,54(6):2137-2148.