Pan Zemin, Zheng Weinan, Zhang Jinli, Gao Rui, Li Dongmei, Guo Xiaoqing, Han Hu, Li Feng, Qu Shen, Shao Renfu
Department of Biochemistry and Molecular Biology, School of Medicine, Shihezi University, Xinjiang Endemic and Ethnic Disease and Education Ministry Key Laboratory, Shihezi, Xinjiang 832002, China.
BMC Cancer. 2014 Jun 10;14:417. doi: 10.1186/1471-2407-14-417.
Cervical carcinoma is the second most common cancer and is an important cause of death in women worldwide. CCAAT/enhancer binding proteins (C/EBPs) are a family of transcription factors that regulate cellular differentiation and proliferation in a variety of tissues. However, the role of C/EBPα gene in cervical cancer is still not clear.
We investigated the expression of C/EBPα gene in cervical squamous cell carcinoma. C/EBPα mRNA level was measured by real-time quantitative RT-PCR in cervical cancer tissues and their adjacent normal tissues. C/EBPα protein level was measured by immunohistochemistry. Methylation in the promoter of C/EBPα gene was detected by MALDI TOF MassARRAY. We transfected HeLa cells with C/EBPα expression vector. C/EBPα expression in HeLa cells was examined and HeLa cell proliferation was measured by MTT assay and HeLa cells migration was analyzed by matrigel-coated transwell migration assays.
There were significant difference in C/EBPα protein expression between chronic cervicitis and cervical carcinoma (P < 0.001). CEBPα mRNA level was significantly lower in cervical cancer tissues than in normal cervical tissues (P < 0.01). Methylation of the promoter of CEBPα gene in CpG 5, CpG-14.15, CpG-19.20 were significantly higher in cervical cancer than in normal cervical tissues (P < 0.05, P < 0.01, P < 0.05, respectively). CEBPα pcDNA3.1 construct transfected into HeLa cells inhibited cell proliferation and decreased cell migration.
Our results indicate that reduced C/EBPα gene expression may play a role in the development of cervical squamous cell carcinoma.
宫颈癌是全球女性中第二常见的癌症,也是导致女性死亡的重要原因。CCAAT/增强子结合蛋白(C/EBPs)是一类转录因子家族,可调节多种组织中的细胞分化和增殖。然而,C/EBPα基因在宫颈癌中的作用仍不清楚。
我们研究了C/EBPα基因在宫颈鳞状细胞癌中的表达情况。通过实时定量逆转录聚合酶链反应(RT-PCR)检测宫颈癌组织及其相邻正常组织中C/EBPα mRNA水平。通过免疫组织化学检测C/EBPα蛋白水平。采用基质辅助激光解吸电离飞行时间质谱(MALDI TOF MassARRAY)检测C/EBPα基因启动子的甲基化情况。我们用C/EBPα表达载体转染HeLa细胞。检测HeLa细胞中C/EBPα的表达,并通过MTT法检测HeLa细胞增殖情况,通过基质胶包被的Transwell迁移实验分析HeLa细胞的迁移情况。
慢性宫颈炎和宫颈癌之间C/EBPα蛋白表达存在显著差异(P < 0.001)。宫颈癌组织中CEBPα mRNA水平显著低于正常宫颈组织(P < 0.01)。宫颈癌中CEBPα基因启动子在CpG 5、CpG - 14.15、CpG - 19.20位点的甲基化水平显著高于正常宫颈组织(分别为P < 0.05、P < 0.01、P < 0.05)。将CEBPα pcDNA3.1构建体转染到HeLa细胞中可抑制细胞增殖并减少细胞迁移。
我们的结果表明,C/EBPα基因表达降低可能在宫颈鳞状细胞癌的发生发展中起作用。