摘要
目的:探讨LIN28A和LAMP1在膀胱癌细胞系中表达情况,以及两者之间的关系,推测其可能临床意义及对肿瘤进展的影响。方法:采用RT-PCR检测膀胱癌细胞系LIN28A、LIN28B和LAMP1表达,免疫荧光检测LIN28A和LAMP1二者蛋白表达定位;LIN28A敲减后通过qRT-PCR检测LAMP1的mRNA表达变化。结果:5个癌细胞系T24、UM-UC3、J82、5637和SW780和正常移行上皮细胞系SV-HUC-1均表达LIN28A,其中J82也表达LIN28B;5个癌细胞系均表达LAMP1,SV-HUC-1不表达LAMP1;LIN28A和LAMP1蛋白均定位在胞浆;LIN28A敲减后对LAMP1的mRNA表达变化无明显影响,相应蛋白变化需要进一步验证。结论:4个膀胱癌细胞系T24、5637、UM-UC3和SW780可以用于LIN28A与肿瘤相关的机制研究,而J82可用于LIN28B的机制研究。LIN28A对肿瘤细胞和干细胞的调控方面可能具有相似性,敲减后对其靶点mRNA表达量无明显影响,LAMP1蛋白可能对肿瘤细胞侵袭转移具有抑制作用。
Objective： To investigate the expression of LIN28A and LAMP1 （lysosome-associated membrane glycoprotein 1） in human bladder cancer cell lines, as well as to speculate their possible clinical significance and effect on tumor progression. Method： RT-PCR was performed to detect the expression of LIN28A, LIN28B and LAMP1 in bladder cancer cell lines. Immunofluorescence was used in detecting the protein location of LIN28A and LAMP1 on bladder tissue and cell lines. The change of the level of mRNA in LAMP1 was checked out by qRT- PCR after knockdown of LIN28A in T24 and 5637 cancer cells. Result： LIN28A was detected in all five bladder cancer cell lines including Tz4 , 5637, UM-UC3, J82, SW780 and transitional epithelium cell lines of SV-HUC-1. LIN28B was also expressed in J82. LAMP1 was not expressed in SV-HUC-1 but all expressed in the above five bladder cancer cell lines. LIN28A and LAMP1 proteins were localized in the cytoplasm of T24 and 5637. Knock- down of LIN28A did not obviously affect the expression of mRNA in LAMP1, and corresponding protein changes were also required to verify. Conclusion： Four bladder cancer cell lines including T24, 5637, UM-UC3 and SW780 can be used in study of LIN28A and mechanism of tumorigenesis, and J82 cell line can be used in investigating the mechanism of LIN28B on tumorigenesis. It is speculated that LIN28A has similar regulation on tumor cells prolif- eration and stem cells differentiation. There is not a change in its target of mRNA levels in LAMP1 after knock- down of LIN28A , and LAMP1 may play a role in prohibiting tumor cells from invading and metastasizing.
出处
《临床泌尿外科杂志》
2014年第1期73-78,共6页
Journal of Clinical Urology
基金
国家自然科学基金(编号81270748
81100471
81070530)
2010年度市科技研发资金基础研究计划资助项目(编号JC201005260214A)
深圳市科技计划重点项目(编号201001014)
深圳市科技计划项目(编号201102001
201302042
201302043
201303041)
