摘要
目的 研究人工合成的dsP21-555对前列腺癌细胞株PC-3和LNCaP细胞周期和增殖的影响.方法 合成dsP21-555(实验组)和dsControl(阴性对照组),分别转染至PC-3和LNCaP.使用Real-time PCR及Western blotting分别检测分析前列腺癌细胞转染后的p21 mRNA及p21蛋白的表达情况.流式细胞术检测细胞周期分布,使用MTF实验及集落形成实验检测细胞的活力及增殖能力.结果 转染dsP21-555后PC-3和LNCaP细胞中的p21mRNA水平分别上调至2.90倍(P<0.01)和2.05倍(P<0.01).Western blotting实验结果符合这一趋势.流式细胞术检测显示,转染dsP21-555后,在S期和G2/M期的细胞比例下降,在G0/G1期的细胞比例则增加.MTT实验显示,与dsControl组相比,转染dsP21-555后,PC-3和LNCaP细胞的活力明显降低.集落形成实验显示,dsP21-555组的集落的数量较少,细胞增殖能力降低.结论 人工合成的dsP21-555能明显激活前列腺癌细胞中p21基因的表达,并显著抑制前列腺癌细胞周期的进展和增殖.
Objective To investigate the effect of dsP21-555 on cell cycle and proliferation of prostate cancer cell line PC-3 and LNCaP.Methods dsP21-555 (experimental group) and dsControl (negative control group) were transfected into PC-3 and LNCaP,respectively.Real-time PCR and Western Blotting were used to detect the expression of p21 mRNA and p21 protein in prostate cancer cells after transfection.The cell cycle distribution was detected by flow cytometry.Cell viability and proliferation were measured by MTT assay and colony forming assay.Results The level of p21 mRNA in PC-3 and LNCaP cells was increased to 2.90-fold (P < 0.01) and 2.05-fold (P < 0.01) respectively after transfection with dsP2l-555.Western Blotting results were consistent with this trend.Flow cytometry showed that the percentage of cells in S phase and G2 / M phase decreased after transfection with dsP21-555,and the proportion of cells in G0 / G1 phase increased.MTT assay showed that the viability of PC-3 and LNCaP cells significantly decreased after dsP21-555 transfection compared with dsControl group.The colony formation assay showed that the number of colonies in the dsP21-555 group was fewer and the cell proliferation ability decreased.Conclusions Synthesized dsP21-555 can significantly activate the expression of p21 gene in prostate cancer cells aud significantly inhibit the progression of cell cycle and proliferation ability of prostate cancer cells.
出处
《国际泌尿系统杂志》
2017年第5期649-652,共4页
International Journal of Urology and Nephrology