宫颈癌局部免疫微环境及其相关蛋白表达分析

Analysis of local immune microenvironment and related protein expression in cervical cancer

目的分析高危型人乳头瘤病毒(high risk human papillomavirus,HR-HPV)感染及CD4、CD8在宫颈鳞状细胞癌(cervical squamous cell carcinoma,CSCC)局部免疫微环境中的作用,探讨HR-HPV感染与蛋白激酶Cι(protein kinase Cι,PKCι)以及Yes相关蛋白1(yes-associated protein 1,YAP1)的相关性。方法收集2013年1月至2015年1月中南大学湘雅医学院附属海口医院全子宫术后CSCC、宫颈高级别鳞状上皮内病变(cervical high-grade squamous intraepithelial lesions,CHSIL)、宫颈低级别鳞状上皮内病变(cervical low-grade squamous intraepithelial lesions,CLSIL)以及正常宫颈组织(normal cervical tissue,NCT)的组织学标本共四组,每组80例,共320例。根据各病例在我院就诊时检测的HR-HPV感染的结果,计算各组HR-HPV的感染率。采用免疫组织化学(immunohistochemistry,IHC)法检测各组组织中CD4、CD8及PKCι、YAP1蛋白表达水平,分析宫颈病变患者HR-HPV感染与CD4、CD8、PKCι、YAP1蛋白表达之间的相关性及影响CSCC患者的预后因素。结果与NCT组相比,LSIL组、HSIL组及CSCC组HR-HPV感染率、PKCι、YAPl及CD8阳性率明显升高,CD4阳性率明显降低,组间差异均有统计学意义(χ^(2)值分别为125.20、122.25、126.82、55.77、71.59,P值均<0.05),且PKCι、YAP1、CD8阳性率越高,CD4阳性率越低。宫颈病变严重程度与HR-HPV感染率、CD8及PKCι、YAP1蛋白阳性表达率均呈显著正相关(r值分别为0.60、0.40、0.58、0.59,P值均<0.05),而CD4蛋白阳性表达率与宫颈病变严重程度呈显著负相关(r=-0.46,P<0.05)。Cox多因素分析结果表明,HR-HPV感染、CD8、PKCι、YAP1蛋白高表达以及淋巴结转移为影响CSCC预后的危险因素,CD4高表达为CSCC预后的保护因素。结论CSCC的发病机制与HR-HPV感染密切相关,同时伴随PKCι、YAP1蛋白的过表达及局部免疫微环境失衡,这有可能为研究CSCC的发病机制及其诊治预后提供一定的依据。

Objective To analyze the role of high-risk human papillomavirus(HR-HPV)infection,CD4 and CD8 in the local immune microenvironment of cervical squamous cell carcinoma(CSCC),and to explore the relationship between HR-HPV infection and protein kinase cι(PKCι),Yes-associated protein 1(YAP1).Methods Four groups of histological specimens of CSCC,cervical high-grade squamous intraepithelial lesions(CHSIL),low-grade squamous intraepithelial lesions(CLSIL)and the normal cervical tissue(NCT)were collected from January 2013 to January 2015 in Affiliated Haikou Hospital of Xiangya Medical College,with 80 cases in each group,and 320 cases in total.The infection rate of the HR-HPV type in each group was calculated according to the results of the HR-HPV infection detected when they visited our hospital.The expression levels of CD4,CD8,PKCιand YAP1 in tissues of each group were detected by immunohistochemistry(IHC).The correlation between HR-HPV infection and the expression of CD4,CD8,PKCι,YAP1 proteins in patients with cervical lesions were analyzed,and explored the prognostic factors affecting CSCC patients.Results Compared with NCT group,The rate of HR-HPV infection and the positive rate of CD8,PKCιas well as YAP1 protein expression was significantly increased,while the positive rate of CD4 was significantly decreased in CSCC,CHSIL and CLSIL group(χ^(2)values were 125.20,122.25,126.82,55.77 and 71.59 respectively,all P values<0.05).The higher the positive rate of PKCι,YAP1 and CD8,the lower the positive rate of CD4.The severity of cervical lesions was significantly positively correlated with the infection rate of HR-HPV,CD8,PKCιand YAP1(r values were 0.60,0.40,0.58 and 0.59 respectively,all P values<0.05),while the positive expression rate of CD4 was significantly negatively correlated with the severity of cervical lesions(r=-0.46,P<0.05).Cox multivariate analysis showed that HR-HPV infection,CD8,PKCι,YAP1 protein expression,and lymph node metastasis were risk factors for CSCC prognosis,and the high expression of CD4 was a protective factor for the prognosis of CSCC.Conclusion The pathogenesis of CSCC is closely related to HR-HPV infection,accompanied by overexpression of PKCιand YAP1 proteins and imbalance of local immune microenvironment,which may provide some basis for studying the pathogenesis,diagnosis and prognosis of CSCC.