Qian Jingfeng, Ji Fangxing, Ye Xue, Cheng Hongyan, Ma Ruiqiong, Chang Xiaohong, Shou Chengchao, Cui Heng
Center of Gynecological Oncology, Department of Obstetrics and Gynecology, Peking University People's Hospital, Beijing 100044, China.
Key Laboratory of Carcinogenesis and Translational Research (Ministry of Education/Beijing), Department of Biochemistry and Molecular Biology, Peking University Cancer Hospital & Institute, Beijing 100142, China.
Chin J Cancer Res. 2018 Apr;30(2):282-290. doi: 10.21147/j.issn.1000-9604.2018.02.11.
Ovarian cancer (OC) is one of the leading causes of death for female cancer patients. COC166-9 is an OC-specific monoclonal antibody and we have identified immunoglobulin γ-1 heavy chain constant region (IGHG1) as its antigen. We explore the function of IGHG1 in proliferation, apoptosis and motility of OC cells further in this research.
IGHG1 expression in OC specimens was detected through immunohistochemistry. Real-time quantitative polymerase chain reaction (RT-qPCR) or western blotting assay was used to test IGHG1 expression in OC cells. Viability of OC cells was tested by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Flow cytometry or western blotting assay was used to detect cell cycle and apoptosis. Cellular motility was analyzed by using transwell assay and the markers of epithelial-mesenchymal transition (EMT) were tested through immunoblots.
Although it exerts negligible effect on the viability and apoptosis of OC cells, IGHG1 could promote migration and invasion of malignant cells . Mechanistically, IGHG1 increases the expression of N-cadherin and Vimentin while decreases E-cadherin expression. Additionally, IGHG1 expression in OC specimens is higher relative to the paired normal counterparts. Further analysis demonstrates that the increased IGHG1 expression correlates positively with the lymph node metastasis of OC.
IGHG1 promotes the motility of OC cells likely through executing the EMT program. Increased IGHG1 expression in OC specimens is associated with the lymph node metastasis.
卵巢癌（OC）是女性癌症患者主要的死亡原因之一。COC166-9是一种卵巢癌特异性单克隆抗体，我们已确定免疫球蛋白γ-1重链恒定区（IGHG1）为其抗原。在本研究中，我们进一步探索IGHG1在卵巢癌细胞增殖、凋亡和迁移中的作用。
通过免疫组织化学检测卵巢癌标本中IGHG1的表达。采用实时定量聚合酶链反应（RT-qPCR）或蛋白质免疫印迹法检测卵巢癌细胞中IGHG1的表达。用3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴盐（MTT）法检测卵巢癌细胞的活力。采用流式细胞术或蛋白质免疫印迹法检测细胞周期和凋亡情况。用Transwell实验分析细胞迁移能力，并通过免疫印迹检测上皮-间质转化（EMT）标志物。
尽管IGHG1对卵巢癌细胞的活力和凋亡影响可忽略不计，但它可促进恶性细胞的迁移和侵袭。机制上，IGHG1增加N-钙黏蛋白和波形蛋白的表达，同时降低E-钙黏蛋白的表达。此外，与配对的正常组织相比，卵巢癌标本中IGHG1的表达更高。进一步分析表明，IGHG1表达增加与卵巢癌的淋巴结转移呈正相关。
IGHG1可能通过执行EMT程序促进卵巢癌细胞的迁移。卵巢癌标本中IGHG1表达增加与淋巴结转移有关。
