miR-629对胰腺癌细胞增殖和迁移的影响

Effect of MiR-629 on Proliferation and Migration of Pancreatic Cancer Cells

[目的]探讨miR-629在胰腺癌组织中的表达及其对胰腺癌细胞增殖和迁移的影响。[方法]选取手术切除的胰腺癌组织及其癌旁组织各72例,采用RT-PCR检测miR-629的mRNA表达水平。构建NC-shRNA和miR-629-shRNA慢病毒稳定细胞系,采用CCK-8法检测miR-629对细胞增殖的影响;采用Transwell检测miR-629对细胞迁移的影响;采用双荧光素酶报告基因和Western blot验证miR-629与SIRT3的靶向关系。[结果]与癌旁组织(0.40±0.04)比较,胰腺癌组织中miR-629的mRNA相对表达水平(1.71±0.15)显著增加(t=3.901,P<0.001);与NC-shRNA细胞比较,miR-629-shRNA细胞的增殖能力均显著下降(F=2.719,P<0.001);与NC-shRNA细胞(140.22±19.37)比较,miR-629-shRNA细胞的迁移能力(63.91±7.44)显著下降(t=4.612,P<0.001);荧光素酶报告基因显示,转染SIRT3-WT后,miR-629-shRNA细胞的相对荧光素酶活性(0.33±0.05)较NCshRNA细胞(1.25±0.12)显著增加(t=4.109,P<0.001);western blot结果表明miR-629-shRNA细胞中SIRT3的蛋白表达水平(0.44±0.03)较NC-shRNA细胞(1.31±0.11)明显增加(t=3.692,P<0.001)。[结论] miR-629在胰腺癌组织中高表达,可能通过靶向下调SIRT3的表达促进胰腺癌细胞的增殖和迁移,有望为胰腺癌的临床诊疗提供新的思路。

[Objective] To investigate the expression of miR-629 in pancreatic cancer and its effect on proliferation and migration of pancreatic cancer cells. [Methods] Seventy-two samples of pancreatic cancer tissue and corresponding adjacent tissue were collected,and the expression of miR-629 was detected by RT-PCR. The NC-sh RNA and miR-629-sh RNA lent viral stable human pancreatic cancer PANC1 cell lines were constructed. The effect of miR-629 on cell proliferation was detected by CCK-8 method;the effect of miR-629 on cell migration was analyzed by Transwell. The targeting relationship between miR-629 and SIRT3 was verified by the dual luciferase reporter gene and Western blot.[Results]Compared with adjacent tissues,the relative m RNA expression of miR-629 in pancreatic cancer tissues was significantly increased(0.40 ±0.04 vs 1.71 ±0.15,t =3.901,P <0.001). Compared with NC-sh RNA cells,the proliferation activity of miR-629-sh RNA cells were significantly decreased(F=2.719,P<0.001).Compared with NC-sh RNA transfected PANC1 cells,the migration ability of miR-629-sh RNA transfected PANC1 cellssignificantly decreased(140.22±19.37 vs 63.91±7.447,t=4.612,P<0.001). Luciferase reporter gene showed that the relative luciferase activity of miR-629-sh RNA cells significantly increased compared with NC-sh RNA cellsafter transfection of SIRT3-WT(1.25 ±0.12 vs 0.33 ±0.05,t =4.109,P <0.001). Western blot showed that the expression of SIRT3 in miR-629-sh RNA cellssignificantly higher than that in NC-sh RNA cells(1.31±0.11 vs 0.44±0.03,t=3.692,P<0.001). [Conclusion]The high expression of miR-629 in pancreatic cancer tissues may promote the proliferation and migration of pancreatic cancer cells by down-regulation of SIRT3 expression,which may provide new ideas for the clinical diagnosis and treatment of pancreatic cancer.