Objective: To explore the expression of CCN5 in endometriotic tissues and its impact on proliferation, migration and invasion of human endometrial stromal cells (HESCs).
Methods: We collected ovarian endometriosis samples from 20 women receiving laparoscopic surgery and eutopic endometrium samples from 15 women undergoing IVF-ET for comparison of CCN5 expression. Cultured HESCs were transfected with a recombinant adenovirus Ad-CCN5 for CCN5 overexpression or with a CCN5-specific siRNA for knocking down CCN5 expression, and the changes of cell proliferation, migration and invasion were evaluated using CCK-8 assay, wound healing assay and Transwell chamber assay. RT-qPCR and Western blotting were used to examine the expression levels of epithelial-mesenchymal transition (EMT) markers including E-cadherin, N-cadherin, Snail-1 and vimentin in HESCs with CCN5 overexpression or knockdown.
Results: CCN5 expression was significantly decreased in ovarian endometriosis tissues as compared with eutopic endometrium samples (P < 0.01). CCN5 overexpression obviously inhibited the proliferation, migration and invasion of HESCs, significantly increased the expression of E-cadherin and decreased the expressions of N-cadherin, Snail-1 and vimentin (P < 0.01). CCN5 knockdown significantly enhanced the proliferation, migration and invasion of HESCs and produced opposite effects on the expressions of E-cadherin, N-cadherin, Snail-1 and vimentin (P < 0.01).
Conclusion: CCN5 can regulate the proliferation, migration and invasion of HESCs and thus plays an important role in EMT of HESCs, suggesting the potential of CCN5 as a therapeutic target for endometriosis.
目的: 分析CCN5在卵巢子宫内膜异位囊肿组织中的表达情况，探究其对子宫内膜基质细胞（HESCs）增殖、迁移和侵袭能力的影响。
方法: 收集并比较卵巢子宫内膜异位症患者组织与正常对照组在位子宫内膜组织中CCN5的表达水平；利用重组腺病毒Ad-CCN5构建过表达CCN5的人子宫内膜基质细胞；采用si-RNA构建敲低CCN5的人子宫内膜基质细胞。利用CCK-8实验检测不同表达水平CCN5对HESCs细胞增殖能力的影响；划痕实验、Transwell小室侵袭实验检测不同表达水平CCN5对HESCs细胞迁移与侵袭能力的影响；Western blot检测不同处理组HESCs中上皮-间充质转化（EMT）标记物E-cadherin、Ncadherin，snail-1和vimentin的表达水平。
结果: 卵巢子宫内膜异位组织中CCN5的表达水平较正常对照组在位子宫内膜显著降低（P < 0.01）；过表达CCN5可以抑制HESCs的增殖、迁徙及侵袭能力；EMT标记物E-cadherin表达升高，N-cadherin、Snail-1和Vimentin表达降低，EMT受到显著抑制（P < 0.01）。而敲低CCN5的表达可以显著增强HESCs的增殖、迁移及侵袭（P < 0.01），降低E-cadherin表达，升高N-cadherin、Snail-1和Vimentin表达，促进HESCs发生EMT转化。
结论: CCN5在卵巢子宫内膜异位组织中的表达水平显著降低，可能通过调控HESCs的增殖、迁移与侵袭能力及影响EMT，在卵巢子宫内膜异位症的发生发展中发挥重要作用。
Keywords: CCN5; endometriosis; epithelial-mesenchymal transition; human endometrial stromal cell; invasion; migration; proliferation.