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目的探讨鸦胆子素D对高危型人乳头瘤病毒(HPV)16感染细胞的增殖抑制作用及对HPV16 E6、HPV16 E7mRNA表达的影响。方法以人宫颈鳞状上皮永生化细胞株(Ect1/E6E7)作为HPV16型感染的体外实验模型,而宫颈癌细胞株Caski作为阳性对照,不同浓度的鸦胆子素D分别体外作用于细胞24 h、48 h、72 h,采用噻唑篮(MTT)法测定细胞体外增殖活性,采用Real-time PCR法检测HPV16 E6、HPV16 E7基因mRNA的表达。结果与同期对照组相比,观察组不同浓度的鸦胆子素D作用的细胞生长抑制率(CGIR)均明显增高,另外,与同一浓度作用24 h相比,作用48 h、72 h的CGIR均明显增高,差异均有统计学意义(均P<0.05)。即鸦胆子素D的浓度越高及作用时间越长,对Ect1/E6E7细胞生长的抑制作用越强。Ect1/E6E7细胞中,观察组不同浓度的HPV16 E6、HPV16 E7 mRNA的表达比较差异无统计学意义,两组比较差异亦无统计学意义(均P>0.05)。而随着鸦胆子素D溶液的浓度上升,Caski细胞中HPV16 E6、HPV16 E7 mRNA的表达升高,两组比较差异有统计学意义(均P<0.05)。结论鸦胆子素D可抑制HPV16 E6、HPV16 E7 mRNA表达和HPV16细胞的增殖。
Objective To investigate the inhibitory effect of Brucein D on the proliferation of human papillomavirus (HPV) 16 infected cells and its effect on HPV16 E6 and HPV16 E7 mRNA expression. Methods Human cervical squamous cell immortalized cell line (Ect1 / E6E7) was used as in vitro experimental model of HPV16 infection, while cervical cancer cell line Caski was used as a positive control. Different concentrations of Brucein D were treated in vitro for 24 h, 48 h and 72 h respectively. The cell proliferation activity was measured by MTT method. The mRNA expression of HPV16 E6 and HPV16 E7 was detected by Real-time PCR. Results Compared with the control group at the same time, the cell growth inhibition rate (CGIR) of the observation group was significantly increased at different concentrations of brucellin D. In addition, compared with the same concentration of 24 h, the CGIR of 48 h and 72 h Significantly higher, the differences were statistically significant (P <0.05). Namely, the higher the concentration of brucellin D and the longer the acting time, the stronger the inhibitory effect on the growth of Ect1 / E6E7 cells. Ect1 / E6E7 cells, the observation group of different concentrations of HPV16 E6, HPV16 E7 mRNA expression was no significant difference between the two groups was also no significant difference (all P> 0.05). However, the expression of HPV16 E6 and HPV16 E7 mRNA in Caski cells increased with the concentration of Brucein D solution (P <0.05). Conclusion Brucein D can inhibit the expression of HPV16 E6 and HPV16 E7 mRNA and the proliferation of HPV16 cells.