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目的 研究外源性p16基因对喉癌细胞系Hep 2的作用 ,并探讨 p16基因用于治疗喉癌的可行性。方法 将重组体腺病毒介导的 p16基因转染到人喉癌细胞系Hep 2 ,用免疫组化、打点杂交方法检测 p16基因在细胞转染前后的表达情况 ,应用流式细胞仪 ,细胞DNALadder等方法研究 p16基因对喉癌细胞周期、形态、生长等特性的影响。结果 感染 p16基因的Hep 2细胞内有外源性 p16基因的表达 ,细胞周期明显变化 ,细胞从G1期到S期发生抑制 ,细胞有退行性改变 ,重组体腺病毒能介导外源基因 p16在喉癌Hep 2细胞系中高效表达。重组体腺病毒介导的p16在Hep 2细胞系中表达 ,能抑制Hep 2细胞系的生长。流式细胞仪计数和细胞DNALadder证实p16能诱导喉癌细胞系Hep 2发生凋亡并导致G1期阻滞。结论 p16基因抑制喉癌细胞系Hep 2的生长可能是通过诱导肿瘤细胞凋亡及G1期阻滞而发挥作用
Objective To investigate the effect of exogenous p16 gene on Hep 2 cell line Hep 2 and investigate the feasibility of p16 gene in treatment of laryngeal cancer. Methods The recombinant adenovirus-mediated p16 gene was transfected into human laryngeal carcinoma cell line Hep 2. The expression of p16 gene was detected by immunohistochemistry and dot blot hybridization before and after transfection. Flow cytometry, The effects of p16 gene on the cell cycle, morphology and growth of laryngeal cancer cells were studied. Results The expression of exogenous p16 gene in Hep 2 cells infected with p16 gene significantly changed the cell cycle. The cells were inhibited from G1 phase to S phase, and the cells were degenerated. The recombinant adenovirus could mediate the expression of the exogenous gene p16 High expression in laryngeal Hep 2 cell line. Recombinant adenovirus-mediated expression of p16 in Hep 2 cell lines can inhibit the growth of Hep 2 cell lines. Flow cytometry and cell DNALadder confirmed that p16 induced apoptosis of laryngeal carcinoma cell line Hep 2 and resulted in G1 arrest. Conclusion Inhibition of p16 gene on the growth of Hep 2 laryngeal carcinoma cell line may play a role by inducing tumor cell apoptosis and G1 phase arrest