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Wip1在肾癌中的表达及其与肿瘤转移的相关性和临床意义。

Expression of Wip1 in kidney carcinoma and its correlation with tumor metastasis and clinical significance.

作者信息

Sun G G, Wang Y D, Liu Q, Hu W N

机构信息

Department of Chemoradiotherapy, Tangshan People's Hospital, Tangshan, 063000, China.

出版信息

Pathol Oncol Res. 2015 Jan;21(1):219-24. doi: 10.1007/s12253-014-9811-9. Epub 2014 Jun 28.

Abstract

This study aimed to analyze the expression, clinical significance of proto-oncogene in kidney carcinoma and the biological effect in its cell line by siRNA targeting wild-type p53-induced phosphatase 1 (Wip1). Immunohistochemistry and western blot were respectively used to analyze Wip1 protein expression in 78 cases of kidney cancer and normal tissues to study the relationship between Wip1 expression and clinical factors. Wip1 siRNA was transiently transfected into papillary kidney carcinoma cell by liposome-mediated method and was detected by Quantitative real-time RT-PCR (qRT-PCR) and western blot. MTT assay, cell apoptosis, cell migration and invasion were also conducted as to the influence of the down-regulated expression of Wip1 that might be found on ACHN cells biological effect. The level of Wip1 protein expression was found to be significantly higher in kidney cancer tissue than normal tissues (P < 0.05). There were significant differences between Wip1 expression and lymph node metastasis, clinical stages and tumor differentiation (P < 0.05). Meanwhile, Increased expression of Wip1 was significantly with poor overall survival time by Kaplan-Meier analysis (P < 0.05). qRT-PCR and Western blot showed that ACHN cell transfected Wip1 siRNA had a lower relative expressive content than normal cell (P < 0.05). MTT assay, cell apoptosis, cell cycles demonstrated that ACHN cell transfected Wip1 siRNA had a lower survival fraction, higher cell apoptosis, more percentage of the G0/G1 phases, significant decrease in migration and invasion, and higher P53 and P16 protein expression compared with ACHN cell untransfected Wip1 siRNA (P < 0.05). Wip1 protein was increased in kidney carcinoma, specifically in T stages, lymph node metastasis, clinical stages and tumor differentiation. Wip1 may involved in the biological processes of kidney cancer cell proliferation, apoptosis, and migration and invasion by regulation P53 and P16 protein expression.

摘要

本研究旨在分析原癌基因在肾癌中的表达、临床意义以及通过靶向野生型p53诱导磷酸酶1(Wip1)的小干扰RNA(siRNA)对其细胞系的生物学效应。分别采用免疫组织化学和蛋白质免疫印迹法分析78例肾癌组织及正常组织中Wip1蛋白的表达情况,以研究Wip1表达与临床因素之间的关系。采用脂质体介导法将Wip1 siRNA瞬时转染至乳头状肾癌细胞中,通过实时定量逆转录聚合酶链反应(qRT-PCR)和蛋白质免疫印迹法进行检测。同时进行MTT法检测、细胞凋亡检测、细胞迁移和侵袭检测,以探讨Wip1表达下调对ACHN细胞生物学效应的影响。结果发现,肾癌组织中Wip1蛋白表达水平显著高于正常组织(P<0.05)。Wip1表达与淋巴结转移、临床分期及肿瘤分化之间存在显著差异(P<0.05)。同时,通过Kaplan-Meier分析发现,Wip1表达增加与总体生存时间缩短显著相关(P<0.05)。qRT-PCR和蛋白质免疫印迹结果显示,转染Wip1 siRNA的ACHN细胞相对表达量低于正常细胞(P<0.05)。MTT法检测、细胞凋亡检测及细胞周期检测结果表明,与未转染Wip1 siRNA的ACHN细胞相比,转染Wip1 siRNA的ACHN细胞存活率降低、细胞凋亡增加、G0/G1期细胞比例增多、迁移和侵袭能力显著下降,且P53和P16蛋白表达升高(P<0.05)。Wip1蛋白在肾癌中表达增加,尤其在T分期、淋巴结转移、临床分期及肿瘤分化方面。Wip1可能通过调节P53和P16蛋白表达参与肾癌细胞增殖、凋亡、迁移和侵袭的生物学过程。

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