Wang Ping, Liu Shan, Cheng Bo, Wu Xizhao, Ding Shanshan, Wu Di, Xu Lin, Shi Jianling, Duan Lian, Sun Suozhu
Department of Pathology, General Hospital of the Second Artillery, Beijing 100088, China.
Department of Pathology, General Hospital of the Second Artillery, Beijing 100088, China; E-mail:
Zhonghua Bing Li Xue Za Zhi. 2015 Dec;44(12):884-8.
To investigate the expression of cyclin D1 in cervical intraepithelial neoplasia (CIN) and squamous cell carcinoma and its relationship with human papillomavirus 16 (HPV16) E7 gene expression.
Both SiHa and Hcc94 cell lines were obtained from cervical epithelial cells of squamous cell carcinoma. E6/E7 gene was silent in Hcc94 cell line.Expression levels of cyclin D1 mRNA and protein in CIN and squamous cell carcinoma were detected by QT-PCR and immunohistochemistry (IHC) respectively. SiRNA was constructed for targeting the promoter of HPV16 E7 and then transfected into SiHa cells to establish cm-16 line with stable silencing of E7. Control cell line B3 was obtained by blank plasmid transfection into SiHa cells. RT-PCR and Western blot were used to detect cyclin D1 mRNA and protein expression in the SiHa, B3, and cm-16 cells, respectively.
Cyclin D1 was expressed in the basal cells of normal cervical squamous epithelia and the expression gradually decreased in the progression from CIN1 to CIN3. Squamous cell carcinoma showed negative or scattered expression of cyclin D1 (P<0.05). Both mRNA and protein of cyclin D1 in E7(+) SiHa cells were lower than those in cm-16 and Hcc94 cells.
Squamous cell carcinoma with high HPV E7 expression shows low level of cyclin D1, suggesting that HPV16 E7 gene inhibits the expression of cyclin D1.
探讨细胞周期蛋白D1在宫颈上皮内瘤变（CIN）及鳞状细胞癌中的表达及其与人乳头瘤病毒16（HPV16）E7基因表达的关系。
SiHa和Hcc94细胞系均取自宫颈鳞状细胞癌上皮细胞。Hcc94细胞系中E6/E7基因沉默。分别采用定量聚合酶链反应（QT-PCR）和免疫组织化学（IHC）检测CIN及鳞状细胞癌中细胞周期蛋白D1 mRNA和蛋白的表达水平。构建针对HPV16 E7启动子的小干扰RNA（SiRNA），转染至SiHa细胞，建立E7稳定沉默的cm-16细胞系。通过将空质粒转染至SiHa细胞获得对照细胞系B3。分别采用逆转录聚合酶链反应（RT-PCR）和蛋白质免疫印迹法检测SiHa、B3和cm-16细胞中细胞周期蛋白D1 mRNA和蛋白的表达。
细胞周期蛋白D1在正常宫颈鳞状上皮的基底细胞中表达，在从CIN1进展至CIN3的过程中表达逐渐降低。鳞状细胞癌中细胞周期蛋白D1呈阴性或散在表达（P<0.05）。E7（+）SiHa细胞中细胞周期蛋白D1的mRNA和蛋白水平均低于cm-16和Hcc94细胞。
HPV E7高表达的鳞状细胞癌中细胞周期蛋白D1水平较低，提示HPV16 E7基因抑制细胞周期蛋白D1的表达。
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