单磷酸阿糖腺苷对HPV16感染永生化宫颈上皮细胞株H8生物学影响

Effects of adenosine monophosphate on immortalized cervical epithelial cell line H8 infected with HPV16

目的探讨单磷酸阿糖腺苷对HPV16感染永生化宫颈上皮细胞株H8增殖、凋亡及侵袭的影响。方法分别以0、20、40、60、80μg/mL浓度单磷酸阿糖腺苷处理体外培养HPV16感染永生化宫颈上皮细胞株H8,CKK-8法分别在12、24、48、72 h检测H8细胞增殖情况,筛选合适药物浓度及作用时间;将体外培养HPV16感染的H8细胞随机分为三组:对照组、单磷酸阿糖腺苷组及顺铂组,以CCK-8法检测细胞活力;以流式细胞技术检测细胞凋亡率;以Transwell侵袭实验和细胞划痕实验检测单磷酸阿糖腺苷对H8细胞侵袭能力的影响;以免疫印记法检测凋亡相关蛋白B淋巴细胞瘤-2基因(Bcl-2)、B淋巴细胞瘤因子相关X蛋白(Bax)及侵袭相关蛋白E-cadherin、Vimentin的表达。结果选择60μg/mL的单磷酸阿糖腺苷处理H8细胞48 h来进行H8细胞增殖、凋亡研究,选择单磷酸阿糖腺苷处理H8细胞12 h后检测细胞侵袭转移力。与对照组比较,单磷酸阿糖腺苷组与顺铂组H8细胞活力、H8细胞数目、侵袭能力、Bcl-2、Vimentin表达明显降低,差异有统计学意义(P<0.05),凋亡率、E-cadherin及Bax表达显著增加,差异有统计学意义(P<0.05);单磷酸阿糖腺苷组和顺铂组相比,各指标差异无统计学意义(P>0.05)。结论单磷酸阿糖腺苷能抑制HPV16感染永生化宫颈上皮细胞H8增殖及侵袭转移能力,促进其凋亡。

Objective To investigate the effects of adenosine monophosphate on proliferation,apoptosis and invasion of immortalized cervical epithelial cell line H8 infected with HPV16. Methods HPV16-infected immortalized cervical epithelial cell line H8 was treated with adenosine monophosphate at concentrations of 0,20,40,60 and 80 μg/mL. CKK-8 method was used to detect the proliferation of H8 cells at 12,24,48 and 72 h,and to screen the appropriate drug concentration and action time. H8 cells infected with HPV16 cultured in vitro. were randomly divided into three groups:control group,adenosine monophosphate group and cisplatin group. Cell viability was measured by CCK-8 assay;the apoptotic rate was detected by flow cytometry;transwell invasion test and cell scratch test were used to detect the effect of adenosine monophosphate on the invasion ability of H8 cells,and the expressions of B-cell lymphoma 2(Bcl-2),Bax,Ecadherin and Vimentin were detected by Western blotting. Results Adenosine monophosphate at 60 μg/mL was used to treat H8 cells for 48 hours to study the proliferation and apoptosis of H8 cells,and the H8 cells treated with adenosine monophosphate for 12 hours were used to detect the invasion and metastasis of H8 cells. Compared with the control group,the viability,numbers,invasiveness,Bcl-2 and Vimentin expressions of H8 cells in adenosine monophosphate group and cisplatin group decreased significantly(P<0.05),while the apoptotic rate,E-cadherin and Bax expressions increased significantly(P<0.05). There was no significant difference in each index between adenosine monophosphate group and cisplatin group(P>0.05). Conclusion Adenosine monophosphate could inhibit the proliferation,invasion and metastasis of immortalized cervical epithelial cells H8 infected with HPV16,and promote its apoptosis.