ENDOD1在前列腺癌组织及细胞中的表达及意义

发布时间：2018-07-17 03:15

【摘要】：目的:观察比较核酸内切酶结构域内含蛋白1(endonuclease domain containing1,ENDOD1)在良性前列腺增生和前列腺癌组织中的表达差异;筛选存在ENDOD1特异性低表达的前列腺癌细胞系,继而通过调控该细胞ENDOD1蛋白表达,研究其在前列腺癌细胞中的生物学功能,初步探索ENDOD1基因与前列腺癌发生、进展的联系。方法:利用免疫组化SP法检测20例良性前列腺增生和21例前列腺癌术后标本组织中ENDOD1表达情况;利用RT-qPCR和Western blot方法观察ENDOD1的mRNA和蛋白在前列腺正常上皮细胞和不同类型前列腺癌细胞中的表达差异,筛选出特异性低表达细胞系;构建pCMV-N-Flag-ENDOD1重组质粒,转染前列腺癌细胞株,过表达ENDOD1蛋白,通过MTT法测定调控前后前列腺癌细胞活力的变化,流式细胞术检测细胞周期和凋亡,Transwell实验评价肿瘤细胞迁移和侵袭能力的改变。结果:免疫组化评分的方差分析结果显示ENDOD1表达与前列腺癌Gleason评分呈负性关联;RT-qPCR和Western blot实验结果表明ENDOD1在雄激素非依赖性前列腺癌细胞系PC3和DU145中存在着特异性低表达(P0.05)。同时,MTT实验显示,在DU145细胞中,过表达ENDOD1肿瘤细胞活力显著下降(P0.05);而流式细胞术检测结果表明过表达ENDOD1能够使DU145细胞周期停滞在G_0/G_1期,但细胞凋亡率无明显差异。此外,在Transwell实验中,过表达ENDOD1的DU145细胞迁移和侵袭能力明显下降(P0.05)。结论:ENDOD1在Gleason评分越高的前列腺癌中表达越低,同时在雄激素非依赖性前列腺癌细胞系存在着特异性低表达;而过表达ENDOD1能明显抑制雄激素非依赖性前列腺癌细胞的生长、迁移和侵袭能力。
[Abstract]:Objective: to investigate the difference of expression of endonuclease domain containing protein 1 (endonuclease domain containing 1 EndoD1 in benign prostatic hyperplasia (BPH) and prostate cancer tissues, and to screen prostate cancer cell lines with low ENDOD1 specific expression. By regulating the expression of ENDOD1 protein, the biological function of ENDOD1 protein in prostate cancer cells was studied, and the relationship between ENDOD1 gene and carcinogenesis and progression of prostate cancer was preliminarily explored. Methods: immunohistochemical SP method was used to detect the expression of ENDOD1 in 20 cases of benign prostatic hyperplasia and 21 cases of prostate cancer after operation. The expression of ENDOD1 mRNA and protein in normal prostate epithelial cells and different types of prostate cancer cells were observed by RT-qPCR and Western blot, and the specific low expression cell lines were screened out, and pCMV-N-Flag-ENDOD1 recombinant plasmid was constructed and transfected into prostate cancer cell lines. Overexpression of ENDOD1 protein was used to detect the changes of prostate cancer cell viability before and after regulation by MTT assay. Flow cytometry was used to detect cell cycle and apoptosis transwell assay was used to evaluate the changes of tumor cell migration and invasion ability. Results: the results of ANOVA showed that the expression of ENDOD1 was negatively correlated with Gleason score of prostate cancer. The results of RT-qPCR and Western blot showed that ENDOD1 was specifically low expressed in androgen independent prostate cancer cell lines PC3 and DU145 (P0.05). At the same time, MTT assay showed that overexpression of ENDOD1 decreased significantly in DU145 cells (P0.05), and flow cytometry showed that overexpression of ENDOD1 could make the cell cycle of DU145 cells stagnate at G0 / GS1 phase, but there was no significant difference in apoptosis rate. In addition, the migration and invasion ability of DU145 cells overexpression of ENDOD1 decreased significantly in Transwell experiment (P0.05). Conclusion the higher the Gleason score, the lower the expression of 1: ENDOD1 in prostate cancer, and the lower specific expression in androgen independent prostate cancer cell line, while the overexpression of ENDOD1 can inhibit the growth of androgen independent prostate cancer cells. Migration and invasion ability.
【作者单位】：中山大学附属第三医院泌尿外科;中山大学附属第三医院急诊科;
【基金】：国家自然科学基金资助项目(No.81402111) 广东省自然科学基金资助项目(No.2015A030313031) 广东省医学科研基金资助项目(No.A2016435)
【分类号】：R737.25

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