miR-126对结肠癌细胞生物学行为的影响及其在调控结肠癌EMT转化中的作用
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摘要
目的:观察miR-126在不同转移潜能的人结肠癌细胞系中的表达情况及其对结肠癌细胞增殖、侵袭转移能力的影响并探讨可能的作用机制。方法:采用实时荧光定量PCR检测结肠癌细胞系(SW480、SW620及HCT116)中miR-126的表达量。通过脂质体瞬时转染法将miR-126过表达(miR-126 mimics),并设置阴性对照组,然后采用CCK8法检测细胞的增殖能力,细胞划痕实验检测细胞的迁移能力,Transwell侵袭小室实验检测细胞的侵袭能力,Western blot实验检测E-cadherin和Vimentin蛋白表达量的变化。结果:相对于低转移潜能的结肠癌细胞株SW480,miR-126在高转移潜能的SW620和HCT116细胞中的表达降低。过表达miR-126可使SW620细胞增殖、迁移和侵袭能力降低,E-cadherin蛋白表达增加,Vimentin蛋白表达降低,差异具有统计学意义(P<0.05)。结论:低表达的miR-126与结肠癌的转移密切相关,miR-126影响结肠癌细胞生物学行为的作用可能是通过调控EMT进程实现的。
Objective:To investigate the expression of miR-126 in human colon cancer cell lines with different metastatic potential and its effect on the proliferation,invasion and metastasis of colon cancer cells,and to explore the possible mechanism.Methods:The expression of miR-126 in human colon cancer cell lines(SW480,SW620 and HCT116) was determined by Real-time fluorescence quantitative PCR.miR-126 mimics were transiently overexpressed in SW620 cells throu Igh liposome transfection and the negative control group was set up.The proliferation ability of cells was detected by CCK8 method and the mobility of cells was detected by wound healing assay and Transwell migration and invasion assay.The expression of E-cadherin,Vimentin was determined by Western blot.Results:The expression of miR-126 was decreased in SW620 and HCT116 cells with high metastatic potential compared SW480 cells with low metastatic potential.The overexpression of miR-126 significantly inhibited the proliferation,migration and invasion ability of SW620 cells and Western blot indicated that miR-126 overexpression increased the expression of E-cadherin and decreased the expression of Vimentin in SW620 cells,which was significantly different from that of negative control(P<0.05).Conclusion:Low expression of miR-126 is closely related to metastasis of colon cancer and the effect of miR-126 on the biological behavior of colon cancer cells may be mediated by the regulation of the EMT process.
Objective:To investigate the expression of miR-126 in human colon cancer cell lines with different metastatic potential and its effect on the proliferation,invasion and metastasis of colon cancer cells,and to explore the possible mechanism.Methods:The expression of miR-126 in human colon cancer cell lines(SW480,SW620 and HCT116) was determined by Real-time fluorescence quantitative PCR.miR-126 mimics were transiently overexpressed in SW620 cells throu Igh liposome transfection and the negative control group was set up.The proliferation ability of cells was detected by CCK8 method and the mobility of cells was detected by wound healing assay and Transwell migration and invasion assay.The expression of E-cadherin,Vimentin was determined by Western blot.Results:The expression of miR-126 was decreased in SW620 and HCT116 cells with high metastatic potential compared SW480 cells with low metastatic potential.The overexpression of miR-126 significantly inhibited the proliferation,migration and invasion ability of SW620 cells and Western blot indicated that miR-126 overexpression increased the expression of E-cadherin and decreased the expression of Vimentin in SW620 cells,which was significantly different from that of negative control(P<0.05).Conclusion:Low expression of miR-126 is closely related to metastasis of colon cancer and the effect of miR-126 on the biological behavior of colon cancer cells may be mediated by the regulation of the EMT process.
引文
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[2]Rodriguez JA,Huerta-Yepez S,Law IK,et al.Diminished expression of CRHR2 in human colon cancer promotes tumor growth and EMT via persistent IL-6/Stat3 signaling[J].Cell Mol Gastroenterol Hepatol,2015,1(6):610-630.
[3]Uwafuji S,Goi T,Naruse T,et al.Protein-bound polysaccharide Kreduced the invasive ability of colon cancer cell lines[J].Anticancer Res,2013,33(11):4841-4845.
[4]Zhu C,Li J,Cheng G,et al.miR-154 inhibits EMT by targeting HMGA2 in prostate cancer cells[J].Mol Cell Biochem,2013,379(1-2):69-75.
[5]He M,Zhan M,Chen W,et al.MiR-143-5p deficiency triggers EMT and metastasis by targeting HIF-1αin gallbladder cancer[J].Cell Physiol Biochem,2017,42(5):2078-2092.
[6]Barbáchano A,Fernández-Barral A,Pereira F,et al.SPROUTY-2represses the epithelial phenotype of colon carcinoma cells via upregulation of ZEB1 mediated by ETS1 and miR-200/miR-150[J].Oncogene,2016,35(23):2991-3003.
[7]Asakura T,Yamaguchi N,Ohkawa K,et al.Proteasome inhibitorresistant cells cause EMT-induction via suppression of E-cadherin by miR-200 and ZEB1[J].Int J Oncol,2015,46(5):2251-2260.
[8]Paterson EL,Kazenwadel J,Bert AG,et al.Down-regulation of the miRNA-200 family at the invasive front of colorectal cancers with degraded basement membrane indicates EMT is involved in cancer progression[J].Neoplasia,2013,15(2):180-191.
[9]Wang C,Zhou B,Liu M,et al.miR-126-5p restoration promotes cell apoptosis in cervical cancer by targeting Bcl2l2[J].Oncol Res,2017,25(4):463-470.
[10]Huang W,Lin J,Zhang H.miR-126:A novel regulator in colon cancer[J].Biomed Rep,2016,4(2):131-134.
[11]Chen X,Cheng H,Pan T,et al.m TOR regulate EMT through Rho A and Rac1 pathway in prostate cancer[J].Mol Carcinog,2015,54(10):1086-1095.
[12]Liu XL,Zhang XT,Meng J,et al.ING5 knockdown enhances migration and invasion of lung cancer cells by inducing EMT via EGFR/PI3K/Akt and IL-6/STAT3 signaling pathways[J].Oncotarget,2017,8(33):54265-54276.