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目的研究清毒栓及β-榄香烯对宫颈癌细胞系HeLa人类白细胞抗原(human leukocyte antigen,HLA)Ⅰ类抗原表达的调节作用,探讨其抗病毒的作用机制。方法水提醇沉法制备清毒栓提取物。体外培养人类宫颈癌细胞HeLa,分为4组:空白对照组、清毒栓组、β-榄香烯组及干扰素α-2b组。MTT法确定3种药物合适的干预浓度,药物干预48 h后,用流式细胞术检测各组细胞表面HLA-Ⅰ的荧光强度,免疫印迹法分析各组细胞内HLA-Ⅰ的蛋白表达,实时荧光定量PCR检测各组细胞HLA-Ⅰ基因表达。结果与空白对照组相比,3种药物均可上调细胞表面HLA-Ⅰ的蛋白表达(P<0.05);且与干扰素α-2b相比,清毒栓及β-榄香烯的上调作用更为显著(P<0.05);此外,3种药物还可上调细胞内HLA-Ⅰ的蛋白表达。与空白对照组相比,清毒栓及β-榄香烯可使HLA-A、B的基因表达增加,而干扰素α-2b只能上调HLA-A的基因表达。结论清毒栓及β-榄香烯可以上调HLA-Ⅰ蛋白及基因表达,从而加强宫颈癌细胞系HeLa表面的抗原呈递,可能是这2种药物帮助机体清除人乳头瘤病毒(HPV)感染及癌变细胞的机制之一。
Objective To investigate the regulatory effect of Qingdu suppository and β-elemene on the expression of human leukocyte antigen (HLA) class I antigen in cervical cancer cell line HeLa and to explore the mechanism of its anti-virus effect. Methods Water extract alcohol precipitation method Qingdu suppository extract. Human cervical cancer HeLa cells were cultured in vitro and divided into 4 groups: blank control group, Qingduo suppository group, β-elemene group and interferon α-2b group. MTT method to determine the appropriate concentration of three drugs intervention, 48 h after drug intervention, the cell surface fluorescence intensity of HLA-Ⅰ was detected by flow cytometry, Western blot analysis of HLA-Ⅰ protein expression in each group of cells, real-time Fluorescent quantitative PCR was used to detect HLA-Ⅰ gene expression in each group. Results Compared with the blank control group, all the three drugs could up-regulate the expression of HLA-Ⅰ on the cell surface (P <0.05). Compared with interferon α-2b, the up-regulation effect of Qingdu suppository and β-elemene (P <0.05). In addition, the three drugs could up-regulate the expression of HLA-Ⅰ in cells. Compared with the blank control group, Qingdubian and β-elemene increased the expression of HLA-A and B genes, while interferon α-2b only up-regulated HLA-A gene expression. Conclusions Qingdu suppository and β-elemene can up-regulate the expression of HLA-Ⅰ protein and gene, so as to enhance the antigen presentation on the HeLa surface of cervical cancer cell line. These two drugs may help the body to clear the human papillomavirus (HPV) infection and One of the mechanisms of cancerous cells.