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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 combined with lentivirus-mediated RNAi silencing VEGF-C gene on the proliferation of esophageal cancer cells in vitro. Methods VEGF targeting was used to design VEGF-siRNA. The lentiviral vector pCDH-VEGF-shRNA was constructed and transfected into esophageal cancer cell line TE1. The expression of VEGF was detected by RT-PCR and Western Blot The MTT assay was used to detect the inhibitory effect of thalidomide combined with lentivirus-mediated silencing of VEGF gene on TE1 cell proliferation. Results The lentiviral vector pCDH-VEGF-shRNA was successfully constructed and packaged with high-titer lentivirus particles. After transfected into TE1 cells, the expression of VEGF in RNAi group was significantly decreased. MTT assay showed thalidomide and RNAi could inhibit TE1 Cell proliferation, but the effect of thalidomide combined with lentivirus-mediated RNAi inhibition of TE1 proliferation is more obvious. Conclusion Thalidomide combined with lentivirus-mediated RNAi can significantly inhibit the expression of VEGF in esophageal cancer cell line TE1 and play an important role in inhibiting the proliferation of TE1 cells.