论文部分内容阅读
目的探讨体外补充不同剂量叶酸对HPV阴性细胞C33A和HPV16阳性细胞CaSki的细胞增殖及凋亡的影响以及叶酸与HPV16的相互关系。方法倒置显微镜下观察叶酸对两种细胞基本生长状态的改变及形态的变化;采用MTT法和流式细胞术观察不同浓度叶酸(0.1、1.0、10、50、100、500、1 000和2 000μg/ml)对两种宫颈癌细胞的增殖抑制、细胞周期及凋亡情况;采用RT-PCR技术检测CaSki细胞中HPV16 E2/E6 mRNA的表达水平。结果补充叶酸对C33A和CaSki细胞的增殖均有抑制作用,细胞形态发生改变,细胞数量明显减少。随着叶酸浓度的增加,对两种宫颈癌细胞的生长抑制率逐渐上升(C33A:r=0.948,P=0.010;CaSki:r=0.895,P=0.006),G0/G1期细胞比例逐渐增大,细胞凋亡率亦随之升高(C33A:r=0.989,P<0.001;CaSki:r=0.994,P<0.001),呈现线性关系。但对C33A和CaSki两种细胞的增殖抑制及凋亡的影响程度无明显差异。当补充叶酸时,CaSki细胞HPV 16E2和E6基因mRNA的表达水平在对照组、叶酸缺乏组和叶酸实验组之间的差异无统计学意义。结论补充叶酸可抑制宫颈癌细胞的增殖,促进凋亡;未发现在宫颈癌细胞中叶酸与HPV16存在交互作用。
Objective To investigate the effects of different doses of folic acid supplementation on cell proliferation and apoptosis of human papillomavirus-type C (HPV-negative) cells and HPV16-positive cells (CaSki) in vitro and the correlation between folic acid and HPV16. Methods The changes of basic growth state and morphological changes of folic acid were observed under inverted microscope. The effects of folic acid (0.1, 1.0, 10, 50, 100, 500, 1000 and 2000 μg) were observed by MTT assay and flow cytometry / ml) on the proliferation, apoptosis and cell cycle of two kinds of cervical cancer cells. The expression of HPV16 E2 / E6 mRNA in CaSki cells was detected by RT-PCR. Results Folic acid supplementation could inhibit the proliferation of C33A and CaSki cells, and the cell morphology changed. The number of cells decreased significantly. With the increase of folate concentration, the growth inhibition rate of two kinds of cervical cancer cells gradually increased (C33A: r = 0.948, P = 0.010; CaSki: r = 0.895, P = 0.006) , The apoptosis rate also increased (C33A: r = 0.989, P <0.001; CaSki: r = 0.994, P <0.001), showing a linear relationship. However, there was no significant difference in the effect of C33A and CaSki on proliferation and apoptosis. When folic acid was supplemented, the expression levels of HPV 16E2 and E6 mRNA in CaSki cells were not significantly different between the control group, folic acid deficiency group and folic acid experimental group. Conclusions Folic acid supplementation can inhibit the proliferation of cervical cancer cells and promote apoptosis. No interaction between folic acid and HPV16 was found in cervical cancer cells.