清毒栓及β-榄香烯对宫颈癌HeLa细胞HLA-Ⅰ类抗原表达的影响  

Influences of Qingdu Suppository and β-elemene on expression of HLA-Ⅰ antigen in cervical cancer cell line HeLa

作　　者：金哲[1] 曹颖[1] 于妍妍[1] 楼姣英[1] 张冬梅[2] 

机构地区：[1]北京中医药大学东方医院,北京100078 [2]北京中医药大学东直门医院中医内科学教育部重点实验室

出　　处：《北京中医药大学学报》2012年第8期529-534,共6页Journal of Beijing University of Traditional Chinese Medicine

基　　金：国家自然科学基金资助项目(No.30873278)

摘　　要：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 study Qingdu Suppository（QDS） and β-elemene on expression of class Ⅰ antigen of leukocyte antigen（HLA-I） of human cervical cancer cell line HeLa,and investigate the anti-viral mechanisms of QDS and β-elemene.Methods The extract of QDS was prepared by using water extraction-alcohol precipitation method,and human cervical cancer cell line HeLa was cultured in vitro and then divided into control group,QDS group,β-elemene group and interferon α-2b group（IFN α-2b group）.The suitable concentration of QDS,β-elemene and IFN α-2b were determined by using MTT test.After the intervention for 48 hours,the fluorescent intensity of HLA-I on cell surface was detected by using flow cytometry,protein expression of HLA-I inside the cells was analysed by using Western blot method,and gene expression of HLA-I was detected by using real-time fluorescence quantitative polymerase chain reaction（RT-PCR） in all groups.Results Compared with control group,the protein expression of HLA-I on cell surface was up-regulated in other three groups（P0.05）,which was more significant in QDS group and β-elemene group（P0.05） compared with IFN α-2b group.The protein expression of HLA-I inside the cells was up-regulated in three medicinal groups.Compared with control group,the expressions of HLA-A mRNA and HLA-B mRNA increased in QDS group and β-elemene group,and only expression of HLA-A mRNA increased in IFN α-2b group.Conclusion QDS and β-elemene can up-regulate the protein and gene expression of HLA-I to promote the antigen presentation on the surface of human cervical cancer cell line HeLa.It is a potential mechanism of these two medicinals in helping the body eliminate human papillomavirus and cancer cells.

关 键 词：宫颈癌 清毒栓 Β-榄香烯 干扰素Α-2B 人乳头瘤病毒 

分 类 号：R285.5[医药卫生—中药学]