肿瘤浸润相关性微小RNA在膀胱尿路上皮癌中表达的初步研究

The infiltration related miRNAs in bladder urothelial carcinoma

目的筛选及验证膀胱尿路上皮癌中肿瘤浸润相关性微小RNA（miRNA）的表达。方法2009年2月至2010年4月随机收集膀胱尿路上皮癌标本20例，肌层浸润组12例，非浸润组8例。采用miRNA芯片分析筛选出肿瘤浸润相关性miRNA。再随机收集膀胱尿路上皮癌标本22例，浸润组15例，非浸润组7例，购置膀胱尿路上皮癌细胞株EJ、5637、T24、BIU-87等4型，采用RT—PCR法验证miRNA基因芯片结果。结果浸润组与非浸润组有7个差异表达基因，hsa—miR-29c、hsa—miR-200a、hsa．miR．378、hsa，miR-429、hsa—miR-200c、hsa—miR-141等上调，hsa-miR-451下调。选取5个miRNAs的RT—PCR验证结果与基因芯片结果一致。膀胱尿路上皮癌T24细胞株5个miRNAs的验证结果与基因芯片结果一致。结论膀胱尿路上皮癌的浸润、进展与miRNA的差异表达密切相关。

Objective To figure out and verify infiltration related miRNAs in bladder urothelial carcinoma （BUC）. Methods Fresh tissues （20 samples, 12 were infiltrative BUC samples, 8 were non-infiltrative BUC samples） were collected in liquid nitrogen. The total RNA was extracted by using Trizol reagents. RNA quality control; miRNA microarray hybridization; data analysis. Another 22 samples were collected in fresh （15 were infiltrative BUC samples, 7 were non-infiltrative BUC samples） for verifying purpose. 4 types of bladder cancer cell lines were used for the study. BUC cell strain ; total RNA was extracted by Trizol reagents; RNA quality control; RT-PCR and analysis of the data. Results ①In infiltrative BUC group, compared with non-infiltrative BUC group, there were 7 differentially expressed miRNAs： hsa-miR- 29e, hsa-miR-200a, hsa-miR-378, hsa-miR-429, hsa-miR-200e and hsa-miR-141 were up-regulated; hsamiR-451 was down-regulated.②In collected samples, the result of RT-PCR was consistent with miRNA array. ③In bladder cancer cell lines, only the results of T24 were consistent with miRNA array. Conclusion Infiltration of BUC might relate with different expression of miRNAs.