论文部分内容阅读
目的:探讨4.1N蛋白对卵巢透明细胞癌细胞迁移和侵袭能力的调控并初步探讨相关分子机制。方法:应用免疫组化法检测4.1N蛋白在卵巢透明细胞癌组织中的表达缺失状况。建立4.1N稳定转染卵巢透明细胞癌ES-2细胞系,通过划痕实验、Transwell迁移和侵袭实验观察细胞迁移侵袭能力的改变。利用qRT-PCR和Western blot法检测紧密连接相关分子(Claudin-4和ZO-1)、上皮-间质转化(EMT)标记物(E-cadherin、Ncadherin和Vimentin)和相关转录因子(Twist、Slug和ZEB-2)及基质金属蛋白酶(MMP-2和MMP-3)表达水平的变化。结果:与卵巢异位的子宫内膜组织相比,卵巢透明细胞癌中4.1N蛋白表达水平显著降低甚至缺失(P<0.0001)。过表达4.1N后,ES-2细胞的迁移和侵袭能力显著降低,Claudin-4和ZO-1表达上调,Twist、Slug、ZEB-2、MMP-2和MMP-3表达水平下降。结论:4.1N在卵巢透明细胞癌细胞中可能通过调控紧密连接蛋白、部分上皮-间质转化转录因子和基质金属蛋白酶的表达水平,抑制癌细胞的迁移和侵袭能力,进而参与负性调控卵巢透明细胞癌的演进。
Objective: To investigate the regulation of 4.1N protein on the migration and invasion of ovarian clear cell carcinoma cells and to explore the underlying molecular mechanisms. Methods: The expression of 4.1N protein in ovarian clear cell carcinoma tissues was detected by immunohistochemistry. 4.1N stably transfected ovarian clear cell carcinoma ES-2 cell line was established, and the invasion and migration of cells were observed by scratch test, Transwell migration and invasion assay. The expressions of Claudin-4 and ZO-1, E-cadherin (Ncadherin and Vimentin) and related transcription factors (Twist, Slug And ZEB-2) and matrix metalloproteinases (MMP-2 and MMP-3) expression levels. Results: 4.1N protein expression in ovarian clear cell carcinoma was significantly lower or even lower than that in ectopic endometrium (P <0.0001). The overexpression of 4.1N could significantly reduce the migration and invasion ability of ES-2 cells. The expressions of Claudin-4 and ZO-1 were up-regulated and the expressions of Twist, Slug, ZEB-2, MMP-2 and MMP-3 were decreased. CONCLUSION: 4.1N can inhibit the migration and invasion of cancer cells by regulating the expression of tight junction proteins, some epithelial-mesenchymal transition transcription factors and matrix metalloproteinases in ovarian clear cell carcinoma cells, and then participate in the negative regulation of ovarian transparency The progression of cell carcinoma.