Rce1在肾细胞癌中的表达及其通过内质网应激对肾癌细胞786-O凋亡的调节

发布时间：2018-04-21 12:48

本文选题：肾细胞癌 + Rce1　；参考：《重庆医科大学》2017年硕士论文

【摘要】：目的:Rce1(Ras and a-factor-converting enzyme 1)是一种位于内质网(Endoplasmic Reticulum,ER)膜上的驻留蛋白酶,负责绝大多数CAAX类蛋白的加工。内质网应激(Endoplasmic Reticulum Stress,ERS)在肾细胞癌(Renal Cell Carcinoma,RCC)发展过程中起重要作用;然而在肾细胞癌中,Rce1的表达,及其与内质网应激作用机制间的关系尚未得到深入研究。本实验旨在探讨Rce1在肾细胞癌中的表达及其通过ERS诱导肾癌细胞786-O凋亡的分子机制。方法:采用实时荧光定量PCR(q RT-PCR)、蛋白质免疫印记杂交(western blot)及免疫组化技术(Immunohistochemical)检测肾细胞癌组织及相应癌旁组织中Rce1表达情况。采用q RT-PCR和western blot技术检测人胚肾细胞株HEK293及肾癌细胞株786-O、ACHN、A498中Rce1表达差异情况。sh RNA慢病毒干扰载体感染786-O细胞,敲降Rce1表达水平,应用流式细胞术(FCM)及CCK-8检测786-O细胞凋亡和增值改变情况。采用western blot技术检测Rce1敲降前后,786-O细胞凋亡相关蛋白Bax、Bcl-2及内质网应激相关蛋白的变化情况。结果:Rce1的表达水平,相较于肾细胞癌旁组织,在肾细胞癌组织中明显上调,其阳性表达水平与患者临床病理特征密切相关。Rce1在人肾癌细胞株中表达上调,尤其是786-O细胞。慢病毒敲降786-O细胞中Rce1表达后,细胞凋亡增加,抑制被增殖(P0.05)。此外,敲降Rce1后,促凋亡蛋白Bax的表达上调,抗凋亡蛋白Bcl-2的表达下调。进一步研究表明,Rce1的下调也影响内质网应激因子的表达。结论:Rce1在肾细胞癌的发生发展中发挥重要作用。Rce1低表达可以通过内质网应激使肾癌细胞的凋亡增加并抑制肾癌细胞增殖。
[Abstract]:Objective: Rce1Ras and a-factor-converting enzyme 1 is a resident protease located on the endoplasmic reticulum Endoplasmic Reticulum enzyme, which is responsible for the processing of most CAAX proteins. Endoplasmic reticulum stress (ERS) plays an important role in the development of renal cell carcinoma (RCC), but the expression of rce1 in renal cell carcinoma and its relationship with the mechanism of endoplasmic reticulum stress have not been further studied. The purpose of this study was to investigate the expression of Rce1 in renal cell carcinoma and the molecular mechanism of 786-O apoptosis induced by ERS. Methods: the expression of Rce1 in renal cell carcinoma (RCC) and adjacent tissues was detected by real-time quantitative PCR(q RT-PCR, Western blot hybridization and immunohistochemical technique. Q RT-PCR and western blot techniques were used to detect the differential expression of Rce1 in human embryonic kidney cell line HEK293 and renal cell line 786-OHCNN A498. Sh RNA lentivirus interference vector was used to infect 786-O cells, and the Rce1 expression level was reduced. Apoptosis and proliferation of 786-O cells were detected by flow cytometry (FCM) and CCK-8. Western blot technique was used to detect the changes of apoptosis related protein Bcl-2 and endoplasmic reticulum stress-related protein before and after Rce1 knockdown. Results compared with the tissues adjacent to renal cell carcinoma, the expression level of Rce1 was significantly up-regulated, and the positive expression level was closely related to the clinicopathological features of the patients. Rce1 was up-regulated in human renal cell carcinoma cell lines, especially 786-O cells. After lentivirus knock down the expression of Rce1 in 786-O cells, the apoptosis increased and the proliferation was inhibited (P0.05). In addition, the expression of pro-apoptotic protein Bax was up-regulated and the expression of anti-apoptotic protein Bcl-2 was down-regulated after knockout of Rce1. Further studies showed that down-regulation of Rce1 also affected the expression of ER stress factors. Conclusion the low expression of Rce1 plays an important role in the genesis and development of renal cell carcinoma. The low expression of Rce1 can increase the apoptosis and inhibit the proliferation of renal cell carcinoma cells through endoplasmic reticulum stress.
【学位授予单位】：重庆医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R737.11

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