miR-195在食管癌中的表达及其对食管癌细胞株增殖的影响

The expression of miR-195 in esophageal cancer and its effect on proliferation of esophageal cancer cell lines

目的:探讨microRNA-195(miR-195)在食管癌中的表达及其对食管鳞癌细胞Eca109增殖的影响。方法:采用Taqman探针实时定量PCR方法检测10对外科手术食管鳞癌标本miR-195的表达,用阳离子脂质体LipofectamineTM2000将miR-195类似物转染入Eca109细胞,Cell Counting Kit-8(CCK-8)法检测细胞增殖情况,流式细胞仪检测细胞周期,Western blot检测蛋白表达水平。结果:与正常食管上皮相比,食管鳞癌组织miR-195表达明显降低(P<0.05)。在48、72、96 h,miR-195转染组细胞吸光值明显低于对照组(P<0.05),且miR-195转染组处于G0/G1期细胞的百分数明显高于对照组(P<0.05)。Western blot结果显示miR-195转染组Cdc42蛋白水平明显低于对照组(P<0.05)。结论:miR-195可阻滞Eca109细胞从G1期进入S期,抑制其增殖。

Objective：To observe the expression of microRNA-195 （miR-195） in ESCC tissues and the effects of miR-195 on the proliferation of Ecal09 cells. Methods：The expression of miR-195 in ten pairs of ESCC tumor tissues and adjacent normal esophageal tissues after surgical resection was detected by Taqman Real-time PCR. The miR-195 mimics was transfected into Ecal09 cell by using LipofectamineTM 2000. The ability of cell proliferation and cell cycle were detected by Cell Counting Kit-8 （CCK-8）and FACS, respectively. Protein was detected by Western Blot. Results：The miR-195 was down-regulated in ESCC tissues compared with normal esophageal tissues （P 〈 0.05）. After transfected miR-195 mimics,cell proliferation was inhibited by CCK-8 assay and cell cycle was arrested in G0/G1 phase by FACS assay （P 〈 0.01）. Western blot result indicated that Cdc42 protein was reduced after miR-195 mimics transfected （P 〈 0.01）. Conclusion：The miR-195 could induce GI arrest and inhibit proliferation in Eca109.