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目的观察RNA干扰介导PIK3CA基因表达沉默对喉鳞状细胞癌(简称喉癌)细胞增殖和侵袭力的影响,探讨PIK3CA基因作为喉癌基因治疗靶标的可行性。方法构建PIK3CA-shRNA的慢病毒表达载体,在脂质体介导下转染人喉癌细胞Hep-2。采用real-time RT-PCR和Western-blot检测PIK3CA基因的表达,MTT法、细胞克隆形成实验、细胞生长曲线检测细胞生长增殖,Boyden小室模型检测细胞的体外侵袭力。结果表达PIK3CA-shRNA的慢病毒载体构建成功;与对照组相比,实验组细胞PIK3CA mRNA和蛋白表达明显下调,分别达75%和70%(P<0.05);细胞增殖和侵袭力均受到明显抑制,差异显著(P<0.05)。结论靶向PIK3CA基因的RNA干扰可抑制喉癌细胞的增殖和侵袭力,PIK3CA有望成为喉癌基因治疗的新的候选基因。
Objective To investigate the effect of PIK3CA gene silencing on the proliferation and invasiveness of laryngeal squamous cell carcinoma (LSCC) and the feasibility of PIK3CA gene as a target of laryngeal cancer gene therapy. Methods The lentiviral expression vector of PIK3CA-shRNA was constructed and transfected into human laryngeal carcinoma Hep-2 cells by liposome. The expression of PIK3CA gene was detected by real-time RT-PCR and Western-blot. MTT assay, cell clone formation assay and cell growth curve were used to detect the proliferation and proliferation of cells. Boyden chamber was used to detect the invasiveness of PIK3CA cells. Results The lentiviral vector expressing PIK3CA-shRNA was successfully constructed. Compared with the control group, the expression of PIK3CA mRNA and protein in the experimental group was significantly down-regulated by 75% and 70% (P <0.05), respectively. The proliferation and invasiveness of PIK3CA- Inhibition, the difference was significant (P <0.05). Conclusions RNA interference targeting PIK3CA gene can inhibit the proliferation and invasiveness of laryngeal carcinoma cells. PIK3CA is expected to be a new candidate gene for gene therapy of laryngeal cancer.