目的:探讨Sirtinol联合K-ras基因RNA干扰对胰腺癌PANC-1细胞增殖和凋亡的影响.方法:用50μmol/m L的Sirtinol和50 nmol K-ras si RNA处理胰腺癌PANC-1细胞48 h,分为C组(无加药处理),K组(加K-ras si RNA处理),S组(加Sirtinol处理),(K+S)...目的:探讨Sirtinol联合K-ras基因RNA干扰对胰腺癌PANC-1细胞增殖和凋亡的影响.方法:用50μmol/m L的Sirtinol和50 nmol K-ras si RNA处理胰腺癌PANC-1细胞48 h,分为C组(无加药处理),K组(加K-ras si RNA处理),S组(加Sirtinol处理),(K+S)组(加K-ras si RNA和Sirtinol处理);Western blot检测SIRT1蛋白的表达情况,Q-PCR检测K-ras m RNA水平和周期蛋白Cyclin D1 m RNA水平,MTT检测细胞的增殖活力,流式细胞仪检测细胞的凋亡情况.结果:Western blot结果显示相对于C组和K组,S组和(K+S)组中的SIRT1蛋白的表达明显下降;Q-PCR结果显示K组和(K+S)组中的K-ras m RNA水平分别是C组的0.454±0.037、0.413±0.032倍,差异具有统计意义;MTT结果显示C组、K组、S组和(K+S)组的A值分别是0.814±0.025、0.634±0.038、0.613±0.036、0.401±0.019,相对于C组,K组、S组和(K+S)组的A值明显下降,其中(K+S)组最明显;Q-PCR结果显示K组、S组和(K+S)组中的Cyclin D1 m RNA水平分别是C组的0.693倍±0.046倍、0.634倍±0.032倍、0.400倍±0.034倍,差异具有统计学意义,其中(K+S)组下降最明显;流式细胞仪显示C组、K组、S组和(K+S)组的凋亡率分别是4.29%±0.246%、7.4 6 9%±0.4 5 7%、8.2 0 6%±0.4 9 0%、12.272%±0.675%,相对于C组,K组、S组和(K+S)组的凋亡率明显上升,其中(K+S)组最明显.结论:Sirtinol联合K-ras si RNA可以更明显的抑制胰腺癌PANC-1细胞的增殖和促进其凋亡.展开更多
Breast cancer is the most common invasive cancer in women worldwide.Sirtuin 1(SIRT1)has recently been shown to have implications in regulating cancer cell growth and apoptosis.SIRT1 regulates Forkhead box O3a(FOXO3a)b...Breast cancer is the most common invasive cancer in women worldwide.Sirtuin 1(SIRT1)has recently been shown to have implications in regulating cancer cell growth and apoptosis.SIRT1 regulates Forkhead box O3a(FOXO3a)by both inhibiting FOXO3-induced apoptosis and potentiating the ability of FOXO3a to resist oxidative stress.Matrix metalloproteinase 2(MMP2)participates in tumor invasion and metastasis by degrading extracellular matrix.SIRT1 up regulates MMP2 expression by its deacetylation activity.This study aimed to investigate the expression of SIRT1,FOXO3a and MMP2 in breast tissues of women with breast cancer.In addition,the effect of SIRT1 inhibition on both FOXO3a and MMP2 expression in breast cancer(MCF-7)cells was assessed.The expression levels of SIRT1,FOXO3a and MMP2 in the breast tissues were determined by real-time PCR in 60 patients with malignant tumor and in 24 patients with benign tumors.After SIRT1 inhibition,protein levels of SIRT1 and FOXO3a were assessed by Western Blot and levels of MMP2 by ELISA in MCF-7 cells.The expression levels of SIRT1,FOXO3a and MMP2 were significantly higher in breast cancer tissues compared to in benign breast tumor and adjacent normal tissues.SIRT1,MMP2 and FOXO3a expression were associated directly with each other.SIRT1 inhibition suppresses MMP2 and FOXO3a expression compared to control MCF7.Sirtinol(SIRT1 inhibitor)effectively induced inhibition of MMP2 and FOXO3a expression in MCF-7 cells,indicating the promising therapeutic strategy of targeting SIRT1 for breast cancer.展开更多
文摘目的:探讨Sirtinol联合K-ras基因RNA干扰对胰腺癌PANC-1细胞增殖和凋亡的影响.方法:用50μmol/m L的Sirtinol和50 nmol K-ras si RNA处理胰腺癌PANC-1细胞48 h,分为C组(无加药处理),K组(加K-ras si RNA处理),S组(加Sirtinol处理),(K+S)组(加K-ras si RNA和Sirtinol处理);Western blot检测SIRT1蛋白的表达情况,Q-PCR检测K-ras m RNA水平和周期蛋白Cyclin D1 m RNA水平,MTT检测细胞的增殖活力,流式细胞仪检测细胞的凋亡情况.结果:Western blot结果显示相对于C组和K组,S组和(K+S)组中的SIRT1蛋白的表达明显下降;Q-PCR结果显示K组和(K+S)组中的K-ras m RNA水平分别是C组的0.454±0.037、0.413±0.032倍,差异具有统计意义;MTT结果显示C组、K组、S组和(K+S)组的A值分别是0.814±0.025、0.634±0.038、0.613±0.036、0.401±0.019,相对于C组,K组、S组和(K+S)组的A值明显下降,其中(K+S)组最明显;Q-PCR结果显示K组、S组和(K+S)组中的Cyclin D1 m RNA水平分别是C组的0.693倍±0.046倍、0.634倍±0.032倍、0.400倍±0.034倍,差异具有统计学意义,其中(K+S)组下降最明显;流式细胞仪显示C组、K组、S组和(K+S)组的凋亡率分别是4.29%±0.246%、7.4 6 9%±0.4 5 7%、8.2 0 6%±0.4 9 0%、12.272%±0.675%,相对于C组,K组、S组和(K+S)组的凋亡率明显上升,其中(K+S)组最明显.结论:Sirtinol联合K-ras si RNA可以更明显的抑制胰腺癌PANC-1细胞的增殖和促进其凋亡.
文摘Breast cancer is the most common invasive cancer in women worldwide.Sirtuin 1(SIRT1)has recently been shown to have implications in regulating cancer cell growth and apoptosis.SIRT1 regulates Forkhead box O3a(FOXO3a)by both inhibiting FOXO3-induced apoptosis and potentiating the ability of FOXO3a to resist oxidative stress.Matrix metalloproteinase 2(MMP2)participates in tumor invasion and metastasis by degrading extracellular matrix.SIRT1 up regulates MMP2 expression by its deacetylation activity.This study aimed to investigate the expression of SIRT1,FOXO3a and MMP2 in breast tissues of women with breast cancer.In addition,the effect of SIRT1 inhibition on both FOXO3a and MMP2 expression in breast cancer(MCF-7)cells was assessed.The expression levels of SIRT1,FOXO3a and MMP2 in the breast tissues were determined by real-time PCR in 60 patients with malignant tumor and in 24 patients with benign tumors.After SIRT1 inhibition,protein levels of SIRT1 and FOXO3a were assessed by Western Blot and levels of MMP2 by ELISA in MCF-7 cells.The expression levels of SIRT1,FOXO3a and MMP2 were significantly higher in breast cancer tissues compared to in benign breast tumor and adjacent normal tissues.SIRT1,MMP2 and FOXO3a expression were associated directly with each other.SIRT1 inhibition suppresses MMP2 and FOXO3a expression compared to control MCF7.Sirtinol(SIRT1 inhibitor)effectively induced inhibition of MMP2 and FOXO3a expression in MCF-7 cells,indicating the promising therapeutic strategy of targeting SIRT1 for breast cancer.
