目的:研究趋化性细胞因子受体CXCR4短干扰RNA(siRNA)对大肠癌细胞系体外侵袭及增殖能力的影响.方法:利用T7 RNA聚合酶体外合成以CXCR4为靶基因的siRNA,用脂质体转染大肠癌SW480细胞,同时设立空白对照组和无关对照组.于转染后48h采用RT-...目的:研究趋化性细胞因子受体CXCR4短干扰RNA(siRNA)对大肠癌细胞系体外侵袭及增殖能力的影响.方法:利用T7 RNA聚合酶体外合成以CXCR4为靶基因的siRNA,用脂质体转染大肠癌SW480细胞,同时设立空白对照组和无关对照组.于转染后48h采用RT-PCR方法检测CXCR4 mRNA水平,免疫印迹方法检测CXCR4和MT1-MMP的蛋白质水平,Boyden小室模型检测体外侵袭能力的变化,流式细胞术检测细胞周期的分布情况,MTT法测定细胞增殖状况.结果:SW480细胞转染CXCR4 siRNA 48h后,与空白对照和无关对照相比,CXCR4 mRNA水平明显下调(51.53%±6.1% vs 78.4%±3.3%.P<0.01:51.53%±6.1% vs 87.4%±5.3%,P<0.01),CXCR4的蛋白质水平明显降低(47.3%±3.7% vs 107.2%±3.6%,P<0.01;47.3%±3.7% vs 114.7%±4.8%,P<0.01),MT1-MMP蛋白表达水平也明显下降(43.8%±2.5% vs 64.4%±4.4%,P<0.01;43.8%±2.5% vs 67.0%±2.9%,P<0.01),细胞的体外侵袭能力减弱(26.5%±6.1% vs 73.7%±3.4%,P<0.01;26.5%±6.1% vs 64.5%±5.7%,P<0.01),细胞周期的分布无明显差异.在无SDF-1存在的情况下,各组细胞的增殖无明显改变;经SDF-1刺激后,各组细胞增殖增加,但CXCR4 siRNA转染组细胞的增殖显著低于空白对照组和无关对照组(24h:0.55±0.03 vs 0.68±0.06,0.71±0.04,P<0.05;48h:0.67±0.04 vs 0.89±0.03,0.94±0.07,P<0.05;72 h:0.72±0.06 vs 1.36±0.08,1.53±0.07,P<0.01).以上各指标在空白对照和无关对照之间无显著性差异(P>0.05).结论:以CXCR4为靶向的siRNA能够有效下调CXCR4基因,降低SDF-1诱导的大肠癌细胞系体外侵袭能力及增殖海性.展开更多
To evaluate the role of salidroside on proliferation,apoptosis and invasiveness of salivary gland adenoid cystic carcinoma cells（SACC）,immunocytochemical staining was employed to detect proliferating cell nuclear an...To evaluate the role of salidroside on proliferation,apoptosis and invasiveness of salivary gland adenoid cystic carcinoma cells（SACC）,immunocytochemical staining was employed to detect proliferating cell nuclear antigen（PCNA）,caspase 3 and caspase 8 expression in SACC-2 cells.Modified Boyden chamber assay combined with laser confocal microscopy（LSCM） was used to evaluate the invasion and migration abilities of SACC-2 cells at different time point.Immunohistochemistry staining revealed that the expression of PCNA was significantly decreased（P0.01） after salidroside treatment.In contrast,salidroside treatment led to increased caspase 3 and caspase 8 in SACC-2 cells.Cell migration depth and number of cells that penetrated Boyden chamber were also decreased by salidroside.Salidroside potently inhibits the proliferation and simultaneously induces the apoptosis of SACC-2 cells.Migration and invasion of SACC-2 cells are also inhibited.Our data throw light on potential clinical application of salidroside to the patients with SACC.展开更多
文摘目的:研究趋化性细胞因子受体CXCR4短干扰RNA(siRNA)对大肠癌细胞系体外侵袭及增殖能力的影响.方法:利用T7 RNA聚合酶体外合成以CXCR4为靶基因的siRNA,用脂质体转染大肠癌SW480细胞,同时设立空白对照组和无关对照组.于转染后48h采用RT-PCR方法检测CXCR4 mRNA水平,免疫印迹方法检测CXCR4和MT1-MMP的蛋白质水平,Boyden小室模型检测体外侵袭能力的变化,流式细胞术检测细胞周期的分布情况,MTT法测定细胞增殖状况.结果:SW480细胞转染CXCR4 siRNA 48h后,与空白对照和无关对照相比,CXCR4 mRNA水平明显下调(51.53%±6.1% vs 78.4%±3.3%.P<0.01:51.53%±6.1% vs 87.4%±5.3%,P<0.01),CXCR4的蛋白质水平明显降低(47.3%±3.7% vs 107.2%±3.6%,P<0.01;47.3%±3.7% vs 114.7%±4.8%,P<0.01),MT1-MMP蛋白表达水平也明显下降(43.8%±2.5% vs 64.4%±4.4%,P<0.01;43.8%±2.5% vs 67.0%±2.9%,P<0.01),细胞的体外侵袭能力减弱(26.5%±6.1% vs 73.7%±3.4%,P<0.01;26.5%±6.1% vs 64.5%±5.7%,P<0.01),细胞周期的分布无明显差异.在无SDF-1存在的情况下,各组细胞的增殖无明显改变;经SDF-1刺激后,各组细胞增殖增加,但CXCR4 siRNA转染组细胞的增殖显著低于空白对照组和无关对照组(24h:0.55±0.03 vs 0.68±0.06,0.71±0.04,P<0.05;48h:0.67±0.04 vs 0.89±0.03,0.94±0.07,P<0.05;72 h:0.72±0.06 vs 1.36±0.08,1.53±0.07,P<0.01).以上各指标在空白对照和无关对照之间无显著性差异(P>0.05).结论:以CXCR4为靶向的siRNA能够有效下调CXCR4基因,降低SDF-1诱导的大肠癌细胞系体外侵袭能力及增殖海性.
基金Supported by the Jilin Provincial Development and Reform Commission,China(No.2007969)the Jilin Provincial Science & Technology Department,China(No.20030551-10)the China Postdoctoral Science Foundation(No.200403699)
文摘To evaluate the role of salidroside on proliferation,apoptosis and invasiveness of salivary gland adenoid cystic carcinoma cells（SACC）,immunocytochemical staining was employed to detect proliferating cell nuclear antigen（PCNA）,caspase 3 and caspase 8 expression in SACC-2 cells.Modified Boyden chamber assay combined with laser confocal microscopy（LSCM） was used to evaluate the invasion and migration abilities of SACC-2 cells at different time point.Immunohistochemistry staining revealed that the expression of PCNA was significantly decreased（P0.01） after salidroside treatment.In contrast,salidroside treatment led to increased caspase 3 and caspase 8 in SACC-2 cells.Cell migration depth and number of cells that penetrated Boyden chamber were also decreased by salidroside.Salidroside potently inhibits the proliferation and simultaneously induces the apoptosis of SACC-2 cells.Migration and invasion of SACC-2 cells are also inhibited.Our data throw light on potential clinical application of salidroside to the patients with SACC.
