华西医学

华西医学

Fibulin-5 基因沉默对人膀胱癌细胞 5637 增殖活力及迁移力的影响

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目的观测 Fibulin-5 基因沉默对人膀胱癌细胞株的生长和迁移能力的影响及其可能的机制。方法采用 Fibulin-5 RNA 干扰慢病毒感染人膀胱癌细胞 5637(F5 组),并设置阴性对照病毒感染的细胞组(NC组)。采用实时荧光定量聚合酶链反应技术检测 Fibulin-5 基因 mRNA 表达水平;M3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴盐法检测 Fibulin-5 基因沉默后 5637 细胞的增殖活性;细胞划痕法测定比较 Fibulin-5 基因沉默对细胞的迁移率的影响;并利用 PathScan 抗体芯片试剂盒检测 Fibulin-5 基因沉默对细胞受体酪氨酸激酶通路蛋白的表达的影响。结果成功建立了 Fibulin-5 沉默细胞株,F5 组的 Fibulin-5 mRNA 表达水平显著低于 NC 组(0.067±0.013 vs. 1.001±0.000),Fibulin-5 基因的敲减效率达到 93.3%;与 NC 组相比,F5 组的细胞增殖活力显著下降,增殖速度明显减缓,同时细胞迁移率均显著下降,差异均有统计学意义(P<0.01);另外,F5 组较 NC 组间变性淋巴瘤激酶、Axl、p44/42 丝裂原活化蛋白激酶、Src 蛋白的表达水平均显著上调(P<0.05)。结论Fibulin-5 可能在膀胱癌细胞的增殖与转移过程中发挥作用,同时可能对胞外信号调节激酶及其信号通路蛋白具有抑制作用。

ObjectiveTo establish human bladder cancer cell line with silenced Fibulin-5 gene and observe the effects and mechanism of Fibulin-5 gene silencing on the proliferation activity and migration of the bladder cancer cells.MethodsThe human bladder cancer cells 5637 were divided into group F5 and group NC, and the cells in group F5 were infected with Fibulin-5 RNA interference (RNAi) lentivirus while the cells in group NC were infected with negative-control virus. Then the expression of Fibulin-5 mRNA was detected by real-time quantitative polymerase chain reaction, the cell proliferation activity was detected by MTT, the migration rate was detected by wound healing method, and the expression levels of proteins in receptor tyrosine kinase (RTK) pathway were detected by PathScan RTK Signaling Antibody Array Kit.ResultsThe Fibulin-5 mRNA expression decreased significantly by Fibulin-5 RNAi lentivirus (0.067±0.013 in group F5 vs. 1.001±0.000 in group NC), and the gene silencing efficiency reached 93.3%, so the Fibulin-5 silencing cell line was established successfully. Comparing with group NC, the relative absorbance value and migration rate of cell 5637 in group F5 decreased significantly (P<0.01); in addition, the expression levels of anaplastic lymphoma kinase, Axl, p44/42 mitogen activated protein kinase, and Src protein were up-regulated in group F5 (P<0.05).ConclusionFibulin-5 may play a role in the proliferation and migration of bladder cancer cells, and may have an inhibitory effect on extracellular signal-regulated kinase and its signaling pathway proteins.

关键词: 膀胱癌细胞; Fibulin-5 基因; 增殖活性; 迁移

Key words: Bladder cancer cells; Fibulin-5 gene; Proliferative activity; Migration

引用本文: 曹贵华, 闫永吉, 陈戬, 张海燕, 王光, 李沛, 石鑫. Fibulin-5 基因沉默对人膀胱癌细胞 5637 增殖活力及迁移力的影响. 华西医学, 2018, 33(4): 417-422. doi: 10.7507/1002-0179.201701037 复制

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