PD-L1在宫颈鳞状细胞癌中的表达及免疫调控作用  

Expression of PD-L1 in cervical squamous cell carcinoma and its immunomodulatory effect

作　　者：陈德慧 杨英捷[2] 王亚菲 孙清华 任婕 Chen Dehui;Yang Yingjie;Wang Yafei;Sun Qinghua;Ren Jie(Clinical College of Medicine,Guizhou Medical University,Guiyang 550004,China;Department of Gynecologic Oncology,Guizhou Cancer Hospital,Guiyang 550004,China;Department of Gynecology,Affiliated Hospital of Guizhou Medical University,Guiyang 550004,China)

机构地区：[1]贵州医科大学临床医学院,贵阳550004 [2]贵州省肿瘤医院妇瘤外科,贵阳550004 [3]贵州医科大学附属医院妇科,贵阳550004

出　　处：《中华实验和临床病毒学杂志》2021年第4期384-388,共5页Chinese Journal of Experimental and Clinical Virology

基　　金：贵州省科教青年英才培养工程[黔省专合字(2012)190号]。

摘　　要：目的揭示宫颈鳞状细胞癌(cervical squamous cell carcinoma,CSCC)组织中E6、E7、PD-L1的表达及相关性,初步探讨E6、E7对PD-L1的免疫调控作用。方法采用Western blot(WB)检测人乳头瘤病毒(human papillomavirus,HPV)阴性正常宫颈组织、宫颈上皮内瘤变(cervical intraepithelial neoplasia,CIN)组织及HPV16、52、58阳性CSCC组织中E6、E7及PD-L1的表达情况,并选取HPV16阳性CSCC组织及HPV阴性CSCC组织进行原代细胞分离、鉴定及培养;构建E6、E7 siRNA,经脂质体转染HPV16阳性CSCC细胞,通过WB检测其细胞中PD-L1表达情况;同时构建E6、E7基因表达质粒,在HPV阴性CSCC细胞中过表达E6、E7,WB检测PD-L1的表达。结果HPV阴性正常宫颈组中未检测到E6、E7及PD-L1的表达,而HPV16、52、58阳性CSCC组中,三者表达均明显高于CIN组(P<0.01);成功分离培养出原代HPV16阳性CSCC细胞及HPV阴性CSCC细胞,分别命名为H16CC细胞株和HNCC细胞株;抑制H16CC细胞株中E6、E7表达后,PD-L1表达明显下降,差异有统计学意义(P<0.05),而过表达HNCC细胞株中E6、E7后,PD-L1表达明显升高,差异有统计学意义(P<0.05)。结论CSCC中PD-L1的表达与E6、E7呈正相关,E6、E7可能通过介导PD-L1的免疫逃逸机制来调控CSCC的发生发展。Objective To disclose the expression and correlation of E6,E7 and PD-L1 in cervical squamous cell carcinoma(CSCC)tissues and explore the immune regulation of E6,E7 on PD-L1.Methods The expressions of E6,E7 and PD-L1 in human papillomavirus(HPV)negative normal cervical tissue,cervical intraepithelial neoplasia(CIN)tissue and HPV16,52 and 58 positive CSCC tissue were detected by Western blot(WB).HPV16 positive CSCC and HPV negative CSCC tissues were used for primary cell isolation,identification and culture.E6,E7 small interfering RNAs(siRNAs)were constructed and transfected into HPV16 positive CSCC cells via liposomes.The expression of PD-L1 in the cells was detected by WB.Plasmids of E6,E7 gene were constructed and overexpressed in HPV negative CSCC cells to detect the expression of PD-L1.Results The expression of E6,E7 and PD-L1 was not detected in the HPV negative normal cervical.While in the HPV16,52,58 positive CSCC tissue group,the expression of all the three were significantly higher than that in the CIN group(P<0.01).Primary HPV16 positive CSCC and HPV negative CSCC cells were successfully isolated and cultured,which were named as H16CC cell lines and HNCC cell lines respectively.After inhibiting the expression of E6,E7 in H16CC cell lines,the expression of PD-L1 was significantly decreased(P<0.05).The expression of PD-L1 was significantly increased after the overexpression of E6,E7 in HNCC cell lines(P<0.05).Conclusions The expression of PD-L1 is positively correlated with E6,E7,which may regulate the occurrence and development of CSCC by mediating the immune escape mechanism of PD-L1 in CSCC.

关 键 词：宫颈鳞状细胞癌 PD-L1 HPV16 免疫调控 

分 类 号：R73[医药卫生—肿瘤]