天津医药

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紫杉醇联合miR-200b对胃腺癌SGC-7901细胞增殖侵袭能力的影响

杜平1,丛宁宁1,申发娟1,康春生2,张庆瑜3   

  1. 1. 天津医科大学总医院
    2. 天津市神经病学研究所脑肿瘤实验室
    3. 天津医科大学总医院消化内科
  • 收稿日期:2012-12-18 修回日期:2013-02-08 出版日期:2013-06-15 发布日期:2013-06-15
  • 通讯作者: 杜平

Effect of paclitaxel combined with miR-200b on proliferation and invasion of gastric cancer cell line SGC-7901

DU PING1,CONG Ningning 1,SHEN Fajuan 1,KANG chunsheng 2,ZHANG Qing yu1   

  1. 1. Department of Gastroenterology, General Hospital of Tianjin Medical University
    2. Laboratory ofNeuro-Oncology, Tianjin Neurological Institute
  • Received:2012-12-18 Revised:2013-02-08 Published:2013-06-15 Online:2013-06-15
  • Contact: DU PING

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摘要： 【摘要】目的 体外观察紫杉醇联合应用miR-200b模拟物（miR-200b mimics）对抑制胃癌SGC-7901细胞增殖侵袭能力的影响。方法  分别单独应用miR-200b转染细胞（200b组）及50 nmol/L紫杉醇单独处理胃癌SGC-7901细胞（紫杉醇组），并联合应用上述两者（联合组），同时设转染无义序列组和对照组。通过流式细胞技术检测细胞的凋亡水平和周期分布情况，克隆形成实验检测细胞的增殖能力，Transwell侵袭实验和细胞划痕实验观测细胞的侵袭迁移能力，Western-blot检测E2F3蛋白的表达水平。结果  与单药组相比，联合组早期凋亡率显著升高；克隆形成率显著降低，穿过Transwell小室的细胞数目显著减少；划痕48 h后的细胞迁移能力明显降低；E2F3蛋白在200b组及联合组中表达水平降低（均P< 0.001）。联合组阻滞于G2/M期细胞的比例高于200b组、无义序列组及对照组，但和紫杉醇组比较差异无统计学意义。结论  miR-200b可能通过降低E2F3表达增强紫杉醇对胃癌SGC-7901细胞增殖侵袭能力的抑制作用。

关键词: 胃肿瘤, 腺癌, 细胞增殖, 紫杉醇, 微RNA-200b

Abstract: Objective    To invesigate the inhibitory effect of paclitaxel combined with miR-200b mimics on proliferation and invasion of human gastric cancer cell line SGC-7901 in vitro. Methods     Human gastric cancer SGC-7901 cells were treated with 50nmol/L paclitaxel alone or combined with transfecting miR-200b mimics. We use the flow cytometry technology, the cell cloning, Transwell assay and wound healing test to observe the change on the the cell cycle, the percentage of cell apoptotic cells, the proliferation ability and the invasive ability. We also discussed the possible mechanism by detecting the level of certain protein using western-blot. Results   The experiments shows that the group treated both with paclitaxel and miR-200b mimics had significantly higher cell apoptotic rate(P<0.001) and lower percentage of cell cloning（P<0.001）, less trans-membrane cell number(P<0.001)，lower migrating rate after 48 h(P<0.001) than the group treated with paclitaxel alone. While there is no significant in the percentage of cells blocked at G2/M phase. As western-blot showed, the level of E2F3 reduced (P<0.001) affter transfecting the miR-200b mimics. Conclusion    MiR-200b may enhance the inhibitory effect of paclitaxel on cell proliferation and invasion of human gastric cancer cell line SGC-7901 by lowering the level of E2F3.

Key words: stomach neoplasms, adenocarcinoma, Proliferation, Paclitaxel, microRNA-200b