清毒栓提取物对人子宫颈鳞癌细胞免疫逃逸的影响  

Effect of Qingdu Suppository extract on immune escape of cervical SiHa cells

作　　者：李美 楼姣英[2] LI Mei;LOU Jiaoying(Beijing University of Chinese Medicine,Beijing 100029,China;不详)

机构地区：[1]北京中医药大学第二临床医学院,100029 [2]北京中医药大学东方医院生殖中心

出　　处：《环球中医药》2024年第2期232-237,共6页Global Traditional Chinese Medicine

基　　金：国家自然科学基金面上项目(81473515)。

摘　　要：目的 研究清毒栓提取物对人子宫颈鳞癌细胞(SiHa细胞)免疫逃逸的影响,探讨清毒栓提取物治疗宫颈人乳头瘤病毒感染的免疫机制。方法 选择均匀成团生长的jurkat细胞,使用SiHa上清液培养jurkat细胞48小时,采用染色质免疫沉淀技术检测SiHa细胞上清液处理后jurkat细胞中叉状头/翅膀状螺旋转录因子(Forkhead boxP3,Foxp3)含量。取Foxp3高表达的T细胞模型,用血清药理学方法制备的清毒栓含药血清和对照血清进行干预,采用染色质免疫沉淀技术、实时荧光定量PCR法、流式细胞术分别检测jurkat细胞中Foxp3蛋白表达水平、Foxp3转录水平,以及Foxp3阳性细胞比例。结果 SiHa上清液可以显著增加jurkat细胞中Foxp3含量(P<0.05),且处理48小时效果最为显著;jurkat细胞经清毒栓含药血清处理后,细胞中的Foxp3蛋白表达显著下调(P<0.05);在含药血清浓度为15%和20%时,Foxp3的蛋白表达量最低,但两者无明显差别(P>0.05);清毒栓含药血清可以显著降低jurkat细胞模型中Foxp3转录和蛋白水平以及Foxp3阳性细胞数(P<0.05)。结论 清毒栓含药血清通过抑制jurkat T细胞中的Foxp3转录和蛋白表达,降低Foxp3阳性细胞数及Treg细胞比例,从而抑制宫颈SiHa细胞的免疫逃逸,增加机体对宫颈SiHa细胞的免疫反应性。Objective To investigate the effect of Qingdu Supposite extract on immune escape of human cervical squamous cell carcinoma(SiHa)cells,and the immune mechanism of Qingdu Suppository extract in the treatment of cervical HPV infection.Methods Jurkat cells grown in homogeneous clusters were first cultured with SiHa supernatant for 48 hours.The expression of Forkhead boxP3(Foxp3)in jurkat cells treated with SiHa supernatant was detected by the method of Western Blot.Next,the T cell model with high Foxp3 expression was used for intervention with Qingdu Supposus-containing serum and control serum prepared by serum pharmacological method.The methods of Western Blot,Quantitative Real-time PCR(qRT-PCR)and flow cytometry were used to detect the expression level of Foxp3 protein and Foxp3 transcription in jurkat cells,respectively.And the proportion of Foxp3 positive cells.Results SiHa supernatant could significantly increase Foxp3 content in jurkat cells(P<0.05),and the effect was most significant after 48h treatment.The Foxp3 protein expression in jurkat cells was significantly down-regulated after treatment with drug-containing serum of Qingdu Suppository(P<0.05),and the Foxp3 protein expression was the lowest when the drug-containing serum concentration was 15%and 20%,and there was no significant difference between the two(P>0.05).The drug-containing serum of Qingdu Suppository could significantly inhibit Foxp3 transcription and protein levels and the number of Foxp3 positive cells in jurkat cell model(P<0.05).Conclusion The drug-containing serum of Qingdu Suppository inhibited Foxp3 transcription and protein expression in jurkat T cells,reduced the number of Foxp3 positive cells and the proportion of Treg cells,and thus inhibited the immune escape of cervical SiHa cells and increased the immune reactivity of the body to cervical SiHa cells.

关 键 词：清毒栓含药血清 宫颈人乳头瘤病毒感染 人子宫颈鳞癌细胞 JURKAT细胞 叉状头/翅膀状螺旋转录因子 

分 类 号：R711.74[医药卫生—妇产科学]