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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 investigate the expression of microRNA-195 (miR-195) in esophageal carcinoma and its effect on the proliferation of esophageal squamous carcinoma cell line Eca109. Methods: The expression of miR-195 in 10 surgical specimens of esophageal squamous cell carcinoma was detected by Taqman real-time PCR. MiR-195 was transfected into Eca109 cells using lipofectamineTM2000. Cell Counting Kit-8 (CCK -8) method to detect cell proliferation, cell cycle detection by flow cytometry, Western blot detection of protein expression levels. Results: Compared with normal esophageal epithelium, the expression of miR-195 in esophageal squamous cell carcinoma was significantly decreased (P <0.05). At 48, 72 and 96 h, the absorbance of miR-195 transfection group was significantly lower than that of control group (P <0.05), and the percentage of cells in G0 / G1 phase of miR-195 transfection group was significantly higher than that of control group <0.05). Western blot results showed that the level of Cdc42 protein in miR-195 transfected group was significantly lower than that in control group (P <0.05). Conclusion: miR-195 can prevent Eca109 cells from entering into S phase from G1 phase and inhibiting its proliferation.