沙利度胺联合RNAi沉默VEGF基因对食管癌细胞增殖的影响

Effect of Thalidomide in combination with RNAi silencing of VEGF gene on proliferation of esophageal cancer cells

目的研究沙利度胺联合慢病毒介导的RNAi沉默VEGF-C基因的方法对食管癌细胞体外增殖的影响。方法以VEGF为靶点,利用siRNA设计原则,设计VEGF-siRNA,构建慢病毒载体pCDH-VEGF-shRNA,并包装好病毒稳定转染到食管癌细胞株TE1中,RT-PCR及Western Blot检测VEGF的表达情况,采用四甲摹偶氮唑蓝(MTT)法检测沙利度胺联合慢病毒介导的RNAi沉默VEGF基因对TE1细胞的增殖抑制作用。结果慢病毒载体pCDH-VEGF-shRNA构建成功并包装出高滴度的慢病毒颗粒,转染TE1细胞后,RNAi组的VEGF的表达明显下降,MTT结果显示沙利度胺及RNAi都可抑制TE1细胞的增殖,但沙利度胺联合慢病毒介导的RNAi抑制TE1增殖的效果更明显。结论沙利度胺联合慢病毒介导的RNAi可明显抑制食管癌细胞株TE1中VEGF的表达,在抑制TE1细胞的增殖中起重要作用。

Objective To study the effect of thalidomide in combination with lentivirus-mediated RNAi silencing of the VEGF gene in vitro of proliferation in esophageal cancer cells.Methods VEGF as a target,VEGF-siRNA was designed using siRNA design principles and inserted into lentiviral vector construct pCDH-VEGF-shRNA,then the virus was packaged and stably transfected to TE1 in esophageal cancer cell line,both of RT-PCR and Western Blot techniques were used to detect the expression of VEGF,the cytostasis of thalidomide in combination with lentiviral-mediated RNAi silencing of the VEGF gene in TE1 cell was evaluated by MTT assay.Results Lentiviral vector pCDH-VEGF-shRNA successfully constructed and packed with a high titer,VEGF expression was significantly decreased in RNAi group after transfected TE1 cells,MTT results showed that both thalidomide and RNAi can inhibited the proliferation of TE1 cell,but Thalidomide in combination with lentiviral-mediated RNAi inhibited the proliferation of TE1 cell more effectively.Conclusions Thalidomide in combination with lentivirus-mediated RNA interference effectively suppressed the expression of VEGF in TE1 cells,which may play an important role in inhibiting the TE1cell proliferation.