特异性miR-221抑制剂对结直肠癌细胞增殖及凋亡的影响.pdfVIP

·674 · 南方医科大学学报（J South Med Univ ） 2011;31(4) ·基础研究· 特异性miR-221 抑制剂对结直肠癌细胞增殖及凋亡的影响 王 伟，孙 凯，吴承堂，雷尚通，曾俊杰，伍颖君，李国新 南方医科大学南方医院普通外科，广东 广州 510515 摘要：目的探讨miR-221 在结直肠癌细胞中的表达状况及特异性miR-221 抑制剂对癌细胞增殖和凋亡的影响。方法应 用实时荧光定量PCR（real-time Q-PCR ）检测四种人结肠癌细胞系HT-29、Lovo、SW-480、Caco2 中miRNA-221 表达状况， 并以人脐静脉内皮细胞HUVEC 作为正常对照；设计并合成miR-221、anti-miR-221（微小RNA 抑制剂）寡核苷酸，以脂质 体转染Caco2 细胞系，以Real-time Q-PCR 再次检测转染后细胞中miR-221 表达状况，并通过噻唑蓝（MTT）比色法及流式 细胞仪观察细胞的增殖和凋亡状态。结果与正常细胞相比，4 种人结肠癌细胞系中miR-221 表达均显著增高，差异具有 统计学意义（P0.01）；癌细胞转染anti-miR-221 后，G /G 期细胞比例明显增加，而S期细胞比例显著下降，并可见细胞凋 0 1 亡的发生（P0.01）。结论miR-221 特异性抑制剂可通过抑制细胞增殖同时诱导凋亡而抑制结直肠癌细胞生长，提示 miR-221 作为结直肠癌生物治疗有效靶点有进一步研究价值。 关键词：结直肠癌；miRNA-221 ；增殖；凋亡 中图分类号：R735.3 文献标志码：A 文章编号：1673-4254（2011）04-0674-04 Effect of miR-221-specific inhibitor on the proliferation and apoptosis of human colorectal carcinoma cells WANG Wei, SUN Kai, WU Cheng-tang, LEI Shang-tong, ZENG Jun-jie, WU Ying-jun , LI Guo-xin Department of General Surgery, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China Abstract: Objective To investigate miRNA-221 expression in human colorectal carcinoma (CRC) cells and the effects of miR-221-specific inhibitor on the proliferation and apoptosis of CRC cells. Methods Four human CRC cell lines (HT-29, Lovo, SW-480, and CaCO2) were examined for miRNA-221 expression using real-time Q-PCR. The specific , 2 -methoxy-modified RNA oligonucleotides of miR-221 (anti-miR-221) were synthesized and transfected into Caco2 cells via liposome, and the changes in the expression of miR-221 in the cells were detected by real-ti