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小RNA干扰介导的ADP核糖基化因子6沉默抑制人前列腺癌PC-3细胞的增殖、迁移和侵袭

[Small RNA interference-mediated ADP-ribosylation factor 6 silencing inhibits proliferation, migration and invasion of human prostate cancer PC-3 cells].

作者信息

Shan Xiong-Wei, Lv Shi-Dong, Yu Xiao-Ming, Hu Zheng-Fei, Zhang Jia-Jie, Wang Guang-Fa, Wei Qiang

机构信息

Department of Urology, Nanfang Hospital, Southern Medical University, Guangzhou, 510515, China. E-mail:

出版信息

Nan Fang Yi Ke Da Xue Xue Bao. 2016 Jun;36(6):735-43.

Abstract

OBJECTIVE

To investigate the effects of silencing ADP-ribosylation factor 6 (Arf6) on the proliferation, migration, and invasion of prostate cancer cell line PC-3 and the possible molecular mechanisms.

METHODS

Three Arf6-specific small interfering RNA (siRNA) were transfected into cultured prostate cancer cell line PC-3. Arf6 expression was examined by real-time PCR and Western blotting. MTT assay, wound healing assay, and Transwell migration and invasion assay were used to observe the effect of Arf6 silencing on the proliferation, migration, and invasion ability of PC-3 cells. The levels of phosphorylated extracellular signal-regulated kinase 1/2 (p-ERK1/2), ERK1/2, p-AKT, AKT and Rac1 were detected by Western blotting.

RESULTS

Transfection of siRNA-3 resulted in significantly decreased Arf6 mRNA and protein expression with inhibition rates of (91.88±3.13)% and (86.37±0.57)%, respectively. Arf6 silencing by siRNA-3 markedly suppressed the proliferation, migration and invasion of PC-3 cells and reduced the expression levels of p-ERK1/2 and Rac1.

CONCLUSION

Silencing of Arf6 efficiently inhibits the proliferation, migration, and invasion of PC-3 cells in vitro, and the underlying mechanisms may involve the down-regulation of p-ERK1/2 and Rac1.

摘要

目的

探讨沉默ADP核糖基化因子6(Arf6)对前列腺癌细胞系PC-3增殖、迁移和侵袭的影响及其可能的分子机制。

方法

将3种Arf6特异性小干扰RNA(siRNA)转染至培养的前列腺癌细胞系PC-3中。通过实时荧光定量PCR和蛋白质印迹法检测Arf6表达。采用MTT法、伤口愈合试验以及Transwell迁移和侵袭试验观察沉默Arf6对PC-3细胞增殖、迁移和侵袭能力的影响。通过蛋白质印迹法检测磷酸化细胞外信号调节激酶1/2(p-ERK1/2)、ERK1/2、p-AKT、AKT和Rac1的水平。

结果

转染siRNA-3后,Arf6 mRNA和蛋白表达显著降低,抑制率分别为(91.88±3.13)%和(86.37±0.57)%。siRNA-3沉默Arf6可显著抑制PC-3细胞的增殖、迁移和侵袭,并降低p-ERK1/2和Rac1的表达水平。

结论

沉默Arf6可有效抑制体外PC-3细胞的增殖、迁移和侵袭,其潜在机制可能涉及p-ERK1/2和Rac1的下调。

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