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50nmol/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48h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。
Objective To investigate the inhibitory effect of paelitaxel 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 divided into three groups, miR-200b group （treated with miR-200b alone）, paclitaxel group （treated with 50 nmol/L paclitaxel alone） and combined group （treated with paclitaxel combined with transfecting miR-200b mimics）. Flow cytometry technology was used to detect the apoptosis and the cell cycle in three groups. The cell cloning was used to detect the proliferation ability, transwell assay and wound healing test were used to observe the invasive ability and Western-blot analysis was used to detect the E2F3 protein level in SGC-7901 cells. Results Compared with paclitaxel group and miR-200b group, the apoptotic rate was significantly higher and the percentage of cell cloning was significantly lower in combined group. Also there was less trans-membrane cell number and lower migrating rate after 48 h （P 〈 0.001）. The expression level of E2F3 protein was reduced in miR-200b group and combined group （P 〈 0.001） after transfection with the miR-200b mimics. There was higher percentage of cells blocked at GJM phase in combined group than that of miR-200b group, but no significant difference when compared with paclitaxel group. 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.
出处
《天津医药》
CAS
北大核心
2013年第6期513-516,共4页
Tianjin Medical Journal
基金
国家自然科学基金资助项目(项目编号:81172356)
天津市自然科学基金重点项目(项目编号:10JCZDJC18500)
关键词
胃肿瘤
腺癌
细胞增殖
紫杉醇
微RNA-200b
stomach neoplasms
adenocarcinoma
cell proliferation
paclitaxel
microRNA-200b
