siRNA沉默STMN1对胃癌AGS细胞紫杉醇敏感性的影响

Effects of silencing STMN1 by siRNA on paclitaxel sensitivity of gastric cancer AGS cells

目的利用siRNA沉默STMN1,观察其对胃癌AGS细胞紫杉醇敏感性的影响。方法将STMN1siRNA通过瞬时转染于胃癌AGS细胞,使用定量聚合酶链反应(qPCR)和蛋白免疫印迹(Western blot)实验检验转染效果,通过MTT实验和平板克隆实验观察AGS细胞对紫杉醇敏感性的变化,并且通过Hoechst 33258染料核染色后观察AGS细胞在紫杉醇作用下凋亡反应的差异。结果成功转染STMN1 siRNA并有效抑制了STMN1的表达。转染STMN1 siRNA后AGS细胞对紫杉醇的药物敏感性明显增加,在紫杉醇作用下细胞凋亡增多。结论通过siRNA抑制STMN1表达可以作为有效的干预手段增加AGS细胞对紫杉醇的化疗敏感性,STMN1或有望成为逆转胃癌耐药的一个新靶点。

[Objective] To investigate the effects of silencing STMN1 by siRNA on paclitaxel sensitivity of gastric cancer AGS cells. [Methods] The STMN1 siRNA （siSTMN1） or scramble siRNA （SCR） were tran- sient transfected into AGS cells. The mRNA and protein levels of STMN1 were detected by qPCR and West- em blot in the AGS cells of different groups. In vitro paclitaxel sensitivity of siSTMN1 and SCR transfected AGS cell lines was tested by MTT assay and colony formation assay. Hoechst 33258 nuclear staining assays were used to investigate the effect of silencing STMN1 on the sensitivity of SCR, siSTMN1 transfected AGS cells and nontreated counterparts （NC） under paclitaxel induced apoptosis. [Results] The transient transfection cell lines were successfully established. Both protein and mRNA levels of STMN1 were effectively down-regu- lated in the siSTMN1 transfected AGS cells. Down-regulation of STMN1 significantly enhanced the sensitivity of AGS cells in response to paelitaxel. In addition, AGS- siSTMN1 cells displayed significant apoptosis as as- sessed by Hoechst nuclear staining. [Conclusion] Silencing STMN1 by siRNA could enhance the sensitivity of AGS cells to paclitaxel and thus is an attractive therapeutic target in the chemotherapy in gastric cancer.