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目的子宫内膜异位症是引起育龄妇女不孕的主要疾病之一,研究表明子宫内膜异位症患者在位子宫内膜间质细胞增殖增强,但具体机制不清。我们研究发现半胱氨酸蛋白酶CAPN 7在子宫内膜异位症患者在位子宫内膜组织中表达显著增高,本研究旨在探讨CAPN 7对子宫内膜间质细胞增殖的影响。方法提取正常育龄妇女及子宫内膜异位症患者在位子宫内膜组织总蛋白,通过Western Blotting检测CAPN 7蛋白表达。制备Ad-GFP-CAPN 7重组腺病毒,采用CCK-8增殖试剂盒和Western Blotting结合腺病毒介导的CAPN 7基因过表达及小干扰RNA介导的CAPN 7基因沉默实验,研究CAPN 7对人子宫内膜间质细胞增殖的影响。结果 CAPN 7蛋白在子宫内膜异位症患者在位子宫内膜组织中的表达明显高于正常育龄妇女子宫内膜组织;成功获得滴度为3.5×1010ifu/mL的重组GFP-CAPN 7腺病毒,人子宫内膜间质细胞感染该腺病毒后,可表达带有绿色荧光蛋白的CAPN 7融合蛋白;伴随CAPN 7蛋白表达量的增加子宫内膜间质细胞中CyclinD3蛋白的表达增加,同时显著增加细胞的增殖能力;基因沉默子宫内膜间质细胞中内源性CAPN 7表达后,子宫内膜间质细胞中CyclinD3蛋白的表达降低且增殖能力降低15%。结论 CAPN 7通过调控细胞周期蛋白CyclinD3的表达促进人子宫内膜间质细胞的增殖。
Endometriosis is one of the major causes of infertility in women of childbearing age. Studies have shown that the proliferation of endometrial stromal cells in patients with endometriosis is enhanced, but the specific mechanism is unclear. Our study found that cystatin CAPN 7 expression in endometriosis patients with endometriosis was significantly increased, the purpose of this study is to investigate CAPN 7 on the proliferation of endometrial stromal cells. Methods The total protein in eutopic endometrium from women of normal reproductive age and endometriosis was extracted and the CAPN 7 protein expression was detected by Western Blotting. The recombinant adenovirus Ad-GFP-CAPN7 was used to detect the CAPN 7 gene expression in CAPN-7 cells by CCK-8 proliferation kit and Western Blotting combined with adenovirus-mediated CAPN 7 gene overexpression and small interfering RNA-mediated CAPN 7 gene silencing Endometrial stromal cell proliferation. Results The expression of CAPN7 protein in endometriosis was significantly higher in endometriosis than in normal women of reproductive age. The recombinant GFP-CAPN7 adenovirus with titer of 3.5 × 1010ifu / mL was successfully obtained , The human endometrial stromal cells infected with the adenovirus can express the CAPN 7 fusion protein with green fluorescent protein; accompanied with the increase of CAPN 7 protein expression, the expression of CyclinD3 protein in endometrial stromal cells increased significantly Increase the cell proliferation; Endometrial stromal cells in the gene silencing endogenous CAPN 7 expression, the expression of CyclinD3 protein in endometrial stromal cells decreased and proliferation decreased by 15%. Conclusion CAPN 7 can promote the proliferation of human endometrial stromal cells by regulating the expression of cyclinD3.