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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 in bladder cancer cell lines and the relationship between them, and to infer the possible clinical significance and the impact on tumor progression. Methods: The expressions of LIN28A, LIN28B and LAMP1 were detected by RT-PCR. The expression of LIN28A and LAMP1 were detected by immunofluorescence. The mRNA expression of LAMP1 was detected by qRT-PCR after LIN28A knockdown. Results: LIN28A was expressed in 5 cancer cell lines T24, UM-UC3, J82, 5637 and SW780 and in normal epithelial cell line SV-HUC-1, of which, J82 also expressed LIN28B; 5 cancer cell lines expressed LAMP1, SV- HUC-1 does not express LAMP1; LIN28A and LAMP1 are located in the cytoplasm; LIN28A knockdown has no significant effect on LAMP1 mRNA expression changes, the corresponding protein changes need further verification. CONCLUSION: Four bladder cancer cell lines, T24, 5637, UM-UC3 and SW780, can be used in the study of tumor-related mechanism of LIN28A and J82 can be used in the mechanism study of LIN28B. LIN28A may have similarities in the regulation of tumor cells and stem cells. After knockdown, LIN28A has no obvious effect on the target mRNA expression. LAMP1 protein may have an inhibitory effect on tumor cell invasion and metastasis.