干扰Arf6抑制人前列腺癌细胞系DU145增殖、迁移及侵袭

基础医学与临床 ›› 2019, Vol. 39 ›› Issue (9): 1310-1315.

干扰Arf6抑制人前列腺癌细胞系DU145增殖、迁移及侵袭

武永强¹,杨帆²

1. 西安工会医院
2. 空军军医大学附属唐都医院泌尿外科

收稿日期:2018-10-22 修回日期:2018-12-24 出版日期:2019-09-05 发布日期:2019-09-06
通讯作者: 杨帆 E-mail:yangfan4220@126.com

Interference with ARF6 inhibits proliferation, migration and invasion of human prostate cancer cell line DU145

Received:2018-10-22 Revised:2018-12-24 Online:2019-09-05 Published:2019-09-06

摘要/Abstract

摘要： 目的观察干扰二磷酸腺苷核糖基化因子6（Arf6）对人前列腺癌细胞系DU145增殖、迁移及侵袭的影响，并初步探讨其调控机制。方法脂质体转染法将靶向Arf6的特异性siRNA序列、无效序列转染至DU145细胞，分别设为干扰组和NC组，不做处理为空白组。蛋白免疫印迹（Western blot）法检测siRNA干扰效率；MTT法检测细胞增殖；划痕试验及Transwell试验检测细胞迁移及侵袭；实时荧光定量聚合酶链反应（RT-qPCR）及Western blot检测Arf6、丝裂原活化的细胞外信号调节激酶（MEK）、细胞外信号调节激酶（ERK）、Ras相关C3肉毒素底物1（Rac1）mRNA及蛋白表达、磷酸化MEK（pMEK）、pERK蛋白表达。结果靶向Arf6的特异性siRNA序列干扰活性为（62.67±4.38）%。干扰组不同时刻细胞增殖均显著低于空白组和NC组（P＜0.05）；干扰组细胞迁移率、穿膜细胞数均显著低于空白组及NC组（P＜0.05）；干扰组Arf6、Rac1 mRNA及蛋白相对表达量、pMEK/MEK、pERK/ERK蛋白比值显著低于空白组及NC组（P＜0.05）。结论干扰Arf6可抑制人前列腺癌细胞系DU145增殖、迁移及侵袭能力，可能与下调pMEK、pERK、Rac1蛋白表达、抑制MEK/ERK信号通路有关。

关键词: 二磷酸腺苷核糖基化因子6, 前列腺癌, 增殖, 迁移, 侵袭

Abstract: Objective To observe the effect of interfering adenosine diphosphate ribosylation factor 6 (Arf6) on proliferation, migration and invasion of human prostate cancer cell line DU145, and to explore its regulatory mechanism. Methods DU145 cells were grown in log phase, and specific siRNA sequences and null sequences targeting Arf6 were transfected into DU145 cells by lipofection, which were set as interference group and NC group. Also take no processing as a blank group. Interference efficiency of siRNA was tested by Western blotting (Western blot). The proliferation of each group was detected and compared by MTT. The migration and invasion of each group were detected and compared by scratch test and Transwell test. Arf6, mitogen-activated extracellular signal-regulated kinase (MEK), extracellular signal-regulated kinase (ERK), Ras-associated C3 botulinum substrate 1 (Rac1) mRNA and protein expressions, phosphorylated MEK (pMEK), pERK protein expressions were detected and compared by real-time fluorescence quantitative polymerase chain reaction (RT-qPCR) and Western blot. Results The interference activity of the specific siRNA sequence targeting Arf6 was (62.67±4.38)%. The cell proliferation of the interference group were significantly lower than those of the blank group and the NC group (P<0.05). The cell migration rate and the number of transmembrane cells of the interference group was significantly lower than that of the blank group and the NC group (P<0.05). The relative expressions of Arf6 and Rac1 mRNA and protein, pMEK/MEK and pERK/ERK protein in the interference group were significantly lower than those in the blank group and NC group (P<0.05). Conclusion Interfering with Arf6 can inhibit the proliferation, migration and invasion of human prostate cancer cell line DU145, which may be related to down-regulation of pMEK, pERK, Rac1 protein expression and inhibition of MEK/ERK signaling pathway.

Key words: Adenosine diphosphate ribosylation factor 6, Prostate cancer, Proliferation, Migration, Invasion

武永强杨帆. 干扰Arf6抑制人前列腺癌细胞系DU145增殖、迁移及侵袭[J]. 基础医学与临床, 2019, 39(9): 1310-1315.

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