m1A甲基转移酶hTrm6p/hTrm61p及其相关基因hTrm6/hTrm61在膀胱尿路上皮癌中的表达及意义
Expression and significance of m1A transmethylase, hTrm6p/hTrm61p and its related gene hTrm6/hTrm61 in bladder urothelial carcinoma.
作者信息
Shi Lei, Yang Xiao-Ming, Tang Dong-Dong, Liu Gang, Yuan Pu, Yang Yang, Chang Lian-Sheng, Zhang Li-Rong, Song Dong-Kui
机构信息
Department of Urology, The First Affiliated Hospital of Zhengzhou University Zhengzhou 450052, China.
Department of Urology, The Chinese People's Liberation Army 153 Central Hospital Zhengzhou 450000, China.
出版信息
Am J Cancer Res. 2015 Jun 15;5(7):2169-79. eCollection 2015.
OBJECTIVE
To discuss the expression of hTrm6p/hTrm61p in bladder urothelial carcinoma tissue and its relationship with m1A level in urine, as well as the influences of hTrm6/hTrm61 on the proliferation and apoptosis of cancer cell line on urothelium.
METHODS
m1A levels in urine of 32 patients of bladder urothelial carcinoma and normal people were detected by HPLC/ESI-Q-TOF-MS, hTrm6p/hTrm61p expression levels in cancer tissue and para-carcinoma tissue of the same patient were detected by western blotting, and hTrm61 expressions of cancer cell line T24, 5637, and EJ of bladder urothelium and kidney cell line HEK-293 of human embryo were detected by RT-qPCR. The hTrm61 high-expression cell line is selected to detect the situation of proliferation and apoptosis by CCK-8 and flow cytometry (FCM) after knocking out hTrm61 with siRNA.
RESULTS
m1A level in urine of carcinoma of urinary bladder is significantly higher than that of normal people, and hTrm6p/hTrm61p expression level in cancer tissue is significantly higher than that in para-carcinoma tissue and has linear correlation with m1A level in urine. The hTrm61 expression in cell line 5637 is significantly higher than that of T24, EJ, and HEK293, and apoptosis is significantly affected after hTrm61 is knocked out from 5637 cell line.
CONCLUSION
High-level expression of hTrm6p/hTrm61p is an important reason for high emission of m1A in urine, and hTrm6/hTrm61 promotes the occurrence of carcinoma of urinary bladder.
目的
探讨hTrm6p/hTrm61p在膀胱尿路上皮癌组织中的表达及其与尿液中m1A水平的关系，以及hTrm6/hTrm61对膀胱尿路上皮癌细胞系增殖和凋亡的影响。
方法
采用HPLC/ESI-Q-TOF-MS检测32例膀胱尿路上皮癌患者及正常人尿液中的m1A水平，采用蛋白质免疫印迹法检测同一患者癌组织及癌旁组织中hTrm6p/hTrm61p的表达水平，采用RT-qPCR检测膀胱尿路上皮癌细胞系T24、5637、EJ及人胚肾细胞系HEK-293中hTrm61的表达。选取hTrm61高表达细胞系，用siRNA敲除hTrm61后，采用CCK-8法和流式细胞术检测细胞增殖和凋亡情况。
结果
膀胱癌患者尿液中m1A水平显著高于正常人，癌组织中hTrm6p/hTrm61p表达水平显著高于癌旁组织，且与尿液中m1A水平呈线性相关。细胞系5637中hTrm61表达显著高于T24、EJ及HEK293，敲除5637细胞系中的hTrm61后，细胞凋亡受到显著影响。
结论
hTrm6p/hTrm61p高表达是尿液中m1A高排放的重要原因，hTrm6/hTrm61促进膀胱癌的发生。
Zotero 插件