稳定感染HPV-16 E6-E7的人喉咽鳞癌FaDu细胞生物学特性观察*
Biology characteristic observation of human laryngopharyngeal squamous cell carcinoma FaDu cells stably infected with HPV-16 E6-E7

目的:观察稳定感染HPV-16 E6-E7对FaDu细胞生物学行为的影响和调控。方法:全基因合成HPV-16 E6-E7基因,将其与慢病毒载体pLV-EGFP-C在T4连接酶作用下连接。制备重组HPV-16 E6-E7慢病毒并感染FaDu细胞。将细胞分3组:实验组(重组HPV-16 E6-E7慢病毒稳定感染的FaDu细胞)、载体对照组(慢病毒空载体稳定感染的FaDu细胞)和空白对照组(未感染慢病毒的FaDu细胞)。分别用CCK-8试剂盒检测各组细胞增殖情况,流式细胞术检测细胞周期和凋亡情况,Transwell侵袭实验检测细胞的体外侵袭能力。结果:获得稳定感染重组HPV-16 E6-E7慢病毒的FaDu细胞,荧光显微镜下可见明亮的绿色荧光蛋白EGFP表达。整合HPV-16 E6-E7基因的FaDu细胞增殖能力和侵袭力均增强,细胞凋亡率降低,S期和G2/M期细胞增加,G0/G1期细胞减少(P<0.05)。结论:成功建立了HPV-16 E6-E7基因整合的FaDu细胞。
Aim: To observe the effect of HPV-16 E6-E7 gene integration on biological behavior of FaDu cells.Methods: Recombinant HPV-16 E6-E7 lentivirus was used to infect FaDu cells. CCK-8 was used to detect the cell proliferation. Flow cytometry was used to detected cell cycle and apoptosis. Transwell invasion assay was used to detected invasive ability.Results: Recombinant FaDu cells with infection by HPV-16 E6-E7 lentivirus expressed green fluorescent protein. The proliferation and invasiveness of FaDu cells with integration of HPV-16 E6-E7 enhanced, apoptosis was inhibited,S phase and G2/M phase cells increased, G0/G1 phase cells reduced(P<0.05).Conclusion: HPV-16 E6-E7 gene integration FaDu cell lines with integration of HPV-16 E6-E7 has been successfully established