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目的构建人IKKα、IKKβ基因RNA干扰慢病毒质粒,并观察其对肝癌细胞生长的影响。方法将目的片段亚克隆入慢病毒载体中,包装成病毒后感染人肝癌细胞,检测细胞IKKα、IKKβ的表达,并将感染细胞接种于裸鼠皮下,观察肿瘤生长情况。结果目的片段被成功克隆入载体质粒。pLenti-GFP-IKKα-SiRNA慢病毒液感染的细胞中IKKα蛋白表达明显降低,pLenti-GFP-IKKβ-SiRNA慢病毒液感染的细胞中IKKβ蛋白表达明显降低。实验组裸鼠皮下肿瘤出现晚且生长缓慢。结论成功构建了针对人IKKα、IKKβ基因RNA干扰慢病毒质粒,包装成病毒后感染人肝癌细胞后能抑制其生长。
Objective To construct the RNA interference plasmids of human IKKα and IKKβ gene and observe their effects on the growth of hepatoma cells. Methods The target fragment was subcloned into lentiviral vector and then packaged into a virus to infect human hepatocellular carcinoma cells. The expression of IKKα and IKKβ in the cells was detected, and the infected cells were inoculated subcutaneously in nude mice to observe the tumor growth. Results The target fragment was successfully cloned into vector plasmid. The expression of IKKαprotein in pLenti-GFP-IKKα-SiRNA lentivirus-infected cells was significantly decreased, while the expression of IKKβprotein in pLenti-GFP-IKKβ-SiRNA lentivirus-infected cells was significantly decreased. The experimental group of nude mice subcutaneous tumors appeared late and slow growth. Conclusion The RNA interference lentiviral plasmid targeting human IKKα and IKKβ gene was successfully constructed, which could inhibit the growth of human hepatocellular carcinoma cells after being packaged into virus.