miR-195在食管癌中的表达及其对食管癌细胞株增殖的影响
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江苏省研究生创新工程(JX22613169);江苏省人民医院食管疾病创新团队项目 (CX11)


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

    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。

    Abstract:

    Objective:To observe the expression of microRNA-195(miR-195) in ESCC tissues and the effects of miR-195 on the proliferation of Eca109 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 Eca109 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 G1 arrest and inhibit proliferation in Eca109.

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傅敏根,李 硕,郁婷婷,钱丽娟,高树娟,朱 宏,冯亚东,曹日昇,陈 亮,施瑞华. miR-195在食管癌中的表达及其对食管癌细胞株增殖的影响[J].南京医科大学学报(自然科学版),2013,(5):575-578

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  • 收稿日期:2012-12-17
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  • 在线发布日期: 2013-05-29
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