siRNA特异性沉默ADP核糖基化因子6对前列腺癌PC-3细胞增殖、迁移和侵袭的影响（英文）
发布时间：2018-10-12 15:38
【摘要】：目的研究ADP核糖基化因子6(Arf6)对雄激素非依赖性前列腺癌PC-3细胞株增殖、迁移和侵袭能力的影响并初步探讨其可能的分子作用机制。方法设计合成3条针对不同靶向区域的Arf6特异性si RNA序列,转染细胞后通过real-time PCR和蛋白质印迹法检测其对Arf6的干扰效果,筛选出干扰效果最佳的si RNA序列;通过噻唑盐(MTT)实验、划痕实验、及transwell细胞迁移和侵袭实验观察si RNA干扰Arf6表达对PC-3细胞增殖、迁移和侵袭的影响;蛋白质印迹法检测AKT、p-AKT、ERK1/2、p-ERK1/2和Rac1蛋白表达水平的变化。结果与空白对照组相比,转染阴性对照序列对PC-3细胞内源性Arf6的m RNA和蛋白表达水平无明显影响,3条si RNA序列均能抑制Arf6的表达,其中si RNA-3对PC-3细胞Arf6表达干扰效果最好,Arf6m RNA和蛋白抑制率分别为(91.88±3.13)%和(86.37±0.57)%。si RNA-3干扰Arf6表达抑制PC-3细胞的增殖,且PC-3细胞体外迁移距离和侵袭细胞数较空白和阴性对照组明显减少(P0.05)。蛋白质印迹法检测发现转染si RNA-3的PC-3细胞p-ERK1/2和Rac1表达水平明显降低,而AKT、p-AKT和ERK1/2表达水平较对照组差异无统计学意义。结论 si RNA干扰Arf6表达可显著抑制PC-3细胞的增殖、迁移和侵袭能力,其分子作用机制可能与p-ERK1/2和Rac1表达下调相关。
[Abstract]:Objective to investigate the effects of ADP ribosylation factor 6 (Arf6) on the proliferation, migration and invasion of androgen independent prostate cancer PC-3 cell line and to explore its possible molecular mechanism. Methods three Arf6 specific si RNA sequences targeting different target regions were designed and synthesized. After transfection, real-time PCR and Western blotting were used to detect the interference effect on Arf6. The effects of si RNA interfering Arf6 expression on the proliferation, migration and invasion of PC-3 cells were observed by scratch assay and transwell cell migration and invasion assay, and the changes of AKT,p-AKT,ERK1/2,p-ERK1/2 and Rac1 protein expression levels were detected by Western blot. Results compared with the blank control group, the expression of m RNA and protein of endogenous Arf6 in PC-3 cells was not significantly affected by the transfection of negative control sequence. All the three si RNA sequences could inhibit the expression of Arf6. The inhibitory rates of Arf6m RNA and protein on Arf6 expression in PC-3 cells were (91.88 卤3.13)% and (86.37 卤0.57)%, respectively. Si RNA-3 interfered with Arf6 expression inhibited the proliferation of PC-3 cells, and the migration distance and the number of invasive cells of PC-3 cells in vitro were significantly decreased compared with the blank and negative control groups (P0.05). Western blot analysis showed that the expression of p-ERK1/2 and Rac1 in PC-3 cells transfected with si RNA-3 was significantly decreased, while the expression of AKT,p-AKT and ERK1/2 was not significantly different from that of the control group. Conclusion si RNA interference with Arf6 expression can significantly inhibit the proliferation, migration and invasion of PC-3 cells, and its molecular mechanism may be related to the down-regulation of p-ERK1/2 and Rac1 expression.
【作者单位】： 南方医科大学南方医院泌尿外科;南方医科大学药学院;
【基金】：Supported by Natural Science Foundation of Guangdong Province(S2013010014537) by Science and Technology Planning Project of Guangdong Province(2012B031800263;2014A010107012;2014A020212260)~~
【分类号】：R737.25
本文编号：2266671
[Abstract]:Objective to investigate the effects of ADP ribosylation factor 6 (Arf6) on the proliferation, migration and invasion of androgen independent prostate cancer PC-3 cell line and to explore its possible molecular mechanism. Methods three Arf6 specific si RNA sequences targeting different target regions were designed and synthesized. After transfection, real-time PCR and Western blotting were used to detect the interference effect on Arf6. The effects of si RNA interfering Arf6 expression on the proliferation, migration and invasion of PC-3 cells were observed by scratch assay and transwell cell migration and invasion assay, and the changes of AKT,p-AKT,ERK1/2,p-ERK1/2 and Rac1 protein expression levels were detected by Western blot. Results compared with the blank control group, the expression of m RNA and protein of endogenous Arf6 in PC-3 cells was not significantly affected by the transfection of negative control sequence. All the three si RNA sequences could inhibit the expression of Arf6. The inhibitory rates of Arf6m RNA and protein on Arf6 expression in PC-3 cells were (91.88 卤3.13)% and (86.37 卤0.57)%, respectively. Si RNA-3 interfered with Arf6 expression inhibited the proliferation of PC-3 cells, and the migration distance and the number of invasive cells of PC-3 cells in vitro were significantly decreased compared with the blank and negative control groups (P0.05). Western blot analysis showed that the expression of p-ERK1/2 and Rac1 in PC-3 cells transfected with si RNA-3 was significantly decreased, while the expression of AKT,p-AKT and ERK1/2 was not significantly different from that of the control group. Conclusion si RNA interference with Arf6 expression can significantly inhibit the proliferation, migration and invasion of PC-3 cells, and its molecular mechanism may be related to the down-regulation of p-ERK1/2 and Rac1 expression.
【作者单位】： 南方医科大学南方医院泌尿外科;南方医科大学药学院;
【基金】：Supported by Natural Science Foundation of Guangdong Province(S2013010014537) by Science and Technology Planning Project of Guangdong Province(2012B031800263;2014A010107012;2014A020212260)~~
【分类号】：R737.25
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