Fan S L, Ding L, Ren Z Y, Chen X, Sun X S, Li C C, Liu C L, Jia W L, Li Q L, Wang J T
Department of Epidemiology, School of Public Health, Shanxi Medical University, Taiyuan 030001, China.
Community Health Centre, Shanxi Cardiovascular Hospital, Taiyuan 030001, China.
Zhonghua Liu Xing Bing Xue Za Zhi. 2017 Jan 10;38(1):96-101. doi: 10.3760/cma.j.issn.0254-6450.2017.01.019.
To investigate the effect of ERK1/2 protein expression and HPV16 infection, as well as their interaction in the cervical carcinogenesis. A total of 176 patients, including 34 cases with normal cervix (NC), 26 cases with cervical intraepithelial neoplasm Ⅰ (CIN Ⅰ), 57 cases with cervical intraepithelial neoplasm Ⅱ/Ⅲ (CIN Ⅱ/Ⅲ) and 59 cases with cervical squamous cell carcinoma (SCC), were enrolled from Shanxi Tumor Hospital, Shanxi Maternal and Child Health Center, Jincheng Coal General Hospital from September 2013 to March 2014. The information about their demographic characteristics and risk factors associated with cervical cancer was collected with structural questionnaire, and cervical tissue samples were collected from each subject. HPV16 infection was detected by PCR, and phosphorylation of ERK1/2 (p-ERK1/2) protein expression levels were detected by immunohistochemistry. Meanwhile, cervical cancer cell lines Siha (HPV16 positive) and C33A (HPV negative) were treated with ERK inhibitor U0126 in vitro. Cell proliferation was determined by living cell count, cell cycle and apoptosis was detected by flow cytometry. The HPV16 infection rate (trend (2)=17.99, <0.001) and p-ERK1/2 protein high expression (trend (2)=10.58, =0.001) increased gradually along with the severity of cervix lesions. There was an additive interaction between HPV16 infection and p-ERK protein expression in the CIN Ⅰ, CIN Ⅱ/Ⅲ and SCC groups. Cell experiments showed that after ERK inhibition, the proliferation of the two cells were reduced (Siha: =6.863, <0.001; C33A: =7.092, <0.001) and the apoptosis were increased (Siha: =-5.201, =0.006; C33A: =-4.335, =0.005). After ERK inhibition, the cell proliferation index of Siha (HPV16 positive) was higher than that of C33A (HPV16 negative) ( =7.066, <0.001), but the apoptosis rate of Siha was lower than that of C33A (=-2.431, =0.057). HPV16 infection and the high expression of p-ERK1/2 could increase the risk of cervical cancer. And there might be synergistic actions between the two factors in the progression of cervical cancer. The effect of ERK1/2 activation to HPV16 infection cells might be more significant in the process of cervical cancer cell proliferation and apoptosis.
为研究细胞外信号调节激酶1/2（ERK1/2）蛋白表达及人乳头瘤病毒16型（HPV16）感染及其二者相互作用在宫颈癌发生中的作用。2013年9月至2014年3月，从山西肿瘤医院、山西省妇幼保健院、晋城煤业集团总医院招募了176例患者，其中包括34例宫颈正常（NC）患者、26例宫颈上皮内瘤变Ⅰ级（CINⅠ）患者、57例宫颈上皮内瘤变Ⅱ/Ⅲ级（CINⅡ/Ⅲ）患者和59例宫颈鳞状细胞癌（SCC）患者。采用结构化问卷收集其人口统计学特征及宫颈癌相关危险因素信息，并采集每位受试者的宫颈组织样本。采用聚合酶链反应（PCR）检测HPV16感染情况，采用免疫组织化学法检测ERK1/2蛋白磷酸化（p-ERK1/2）表达水平。同时，体外使用ERK抑制剂U0126处理宫颈癌细胞系Siha（HPV16阳性）和C33A（HPV阴性）。通过活细胞计数测定细胞增殖，采用流式细胞术检测细胞周期和凋亡情况。HPV16感染率（趋势检验（2）=17.99，P<0.001）和p-ERK1/2蛋白高表达率（趋势检验（2）=10.58，P=0.001）随宫颈病变严重程度逐渐升高。在CINⅠ、CINⅡ/Ⅲ和SCC组中，HPV16感染与p-ERK蛋白表达之间存在相加交互作用。细胞实验表明，ERK抑制后，两种细胞的增殖均降低（Siha：t=6.863，P<0.001；C33A：t=7.092，P<0.001），凋亡增加（Siha：t=-5.201，P=0.006；C33A：t=-4.335，P=0.005）。ERK抑制后，Siha（HPV16阳性）细胞增殖指数高于C33A（HPV16阴性）（t=7.066，P<0.001），但Siha细胞凋亡率低于C33A（t=-2.431，P=0.057）。HPV16感染和p-ERK1/2高表达可增加宫颈癌发病风险。二者在宫颈癌进展过程中可能存在协同作用。在宫颈癌细胞增殖和凋亡过程中，ERK1/2激活对HPV16感染细胞的影响可能更显著。
