siRNA沉默胰腺癌Panc-1中THOC1对吉西他滨敏感性的影响

Effects of silencing THOC1 by si RNA on sensitivity of pancreas cancer cells Panc-1 to gemcitabine

目的观察小干扰RNA(si RNA)沉默THOC1对胰腺癌Panc-1细胞吉西他滨敏感性的影响。方法将THOC1 si RNA通过瞬时转染于胰腺癌Panc-1细胞,使用q PCR和Western blot实验检验转染效果,通过MTT实验和平板克隆实验观察Panc-1细胞对吉西他滨敏感性的变化,并且通过Hoechst 33258染料核染色后观察Panc-1细胞在吉西他滨作用下凋亡反应的差异。结果成功转染THOC1 si RNA并有效抑制了THOC1的表达。转染THOC1 si RNA后Panc-1细胞对吉西他滨的药物敏感性明显增加(P<0.01),在吉西他滨作用下凋亡细胞增多(P<0.01)。结论通过si RNA抑制THOC1的表达后可以增加胰腺癌Panc-1细胞对吉西他滨的药物敏感性。

[ObjectIve] To investigate the effects of silencing THOC1 by siRNA on the sensitivity of pancreas cancer cells Panc-I to gemcitabine. [ Methods ] The THOC1 siRNA （THOC1 siRNA） or Control siRNAamble siRNA （Control siRNA） were transient transfected into Panc-1 cells. The mRNA and protein levels of THOC1 were detected by qPCR and Western blot in the Panc-lcells of different groups. In vitro gemcitabine sensitivity of THOC1 siRNA and Control siRNA transfected Panc-lcell lines was tested by MTF assay and colony formation assay. Hoechst 33258 nuclear staining were used to investigate the effect of silencing THOC1 on the sensitivity of Control siRNA, THOCI siRNA transfected Panc-lcells and nontreated counterparts undergemcitabine induced apoptosis. [Results ] The transient transfection cell lines were successfully established. Both protein and mRNA levels of THOC1 were ef- fectively down-regulated in the THOC1 siRNA transfected Panc-lcells. Down-regulation of THOC1 significantly en- hanced the sensitivity of Panc-lcells in response to gemcitabine（P 〈0.01）. In addition, the THOC1 siRNA transfect- ed Panc-1 cells displayed significant apoptosis as assessed by Hoechst nuclear staining （P 〈0.01）. [ Conclusions] Silencing THOC1 by siRNA could enhance the sensitivity of pancreas cancer cells Panc-1 to gemcitabine.