血根碱对紫杉醇耐药卵巢癌A2780/Taxol细胞生长及TGF-β_1/Smad通路抑制的影响
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摘要
目的探讨血根碱对紫杉醇耐药卵巢癌A2780/Taxol细胞生长及TGF-β_1/Smad通路和紫杉醇敏感性的影响。方法将A2780/Taxol细胞分为对照组、血根碱组、紫杉醇组、血根碱联合紫杉醇组(简称联合组),对照组加生理盐水,其余3组分别应用2μmol/L血根碱、2μg/mL紫杉醇,2μmol/L血根碱+2μg/mL紫杉醇处理。48 h后应用倒置显微镜观察A2780/Taxol细胞形态、MTT法检测细胞增殖,流式细胞仪检测细胞周期,Western blot法检测Caspase-3、TGF-β_1、Smad7蛋白表达。结果对照组细胞贴壁生长,边界清楚;血根碱组细胞体积缩小,贴壁数减少;紫杉醇组见细胞呈圆形脱落;联合组上过变化最为明显。与对照组比较,3组均可显著抑制A2780/Taxol细胞增殖,上调Caspase-3、Smad7蛋白表达、下调TGF-β_1蛋白表达,联合组作用最为明显(P<0.05)。血根碱组对细胞周期无影响,紫杉醇组及联合组G0/G1期、S期细胞减少,G2/M期细胞增多。结论血根碱通过抑制TGF-β_1/Smad通路,促进Caspase-3表达抑制A2780/Taxol细胞生长,促进紫杉醇药物敏感性。
Objective To study the effect of Sanguinarine on A2780/Taxol cell growth,TGF-β_1/Smad pathway,and the sensitivity to paclitaxel in treatment of paclitaxel resistant ovarian cancer. Methods A2780/Taxol cells were divided into the control group,the sanguinarine group,the paclitaxel group,the paclitaxel combined sanguinarine group( combined group in short). Normal saline was added to cells in control group. Cells in the rest three groups were added 2. 0 μmol/L sanguinarine,2 μg/mL paclitaxel,and 2. 0μmol/L sanguinarine combined 2 μg/mL paclitaxel respectively. The morphology of A2780/Taxol cells was observed by inverted microscope. The cell proliferation were detected by MTT. The cell cycle was detected by flow cytometry. And the expressions of Caspase-3,TGF-β_1,and Smad7 proteins were detected by Western blot.Results The cells in control group were adhered to the wall and the bounday was clear. the cell volume decreased and the number of adherent cells decreased in sanguinarine group. The cells in Taxol group fall off were circularly. These changes were most obvious in the combined group. Compared with the control group,the proliferation of A2780/Taxol cells could be significantly inhibited,the expressions of Caspase-3 and Smad7 protein were up-regulated,the expression of TGF-β_1 protein was down-regulated in the rest 3 groups. The most effect was obviously seen in the combined group(P < 0. 05). Sanguinarine showed no effect on cell cycle. The cell numbers of G0/G1 and S phase decreased and the cell number of G2/M phase increased in the paclitaxel group and the combined group. Conclusion Sanguinarine could inhibit the growth of A2780/Tax-ol cells and enhance the sensitivity of paclitaxel by suppressing TGF-β_1/Smad pathway and promoting Caspase-3 expression.
Objective To study the effect of Sanguinarine on A2780/Taxol cell growth,TGF-β_1/Smad pathway,and the sensitivity to paclitaxel in treatment of paclitaxel resistant ovarian cancer. Methods A2780/Taxol cells were divided into the control group,the sanguinarine group,the paclitaxel group,the paclitaxel combined sanguinarine group( combined group in short). Normal saline was added to cells in control group. Cells in the rest three groups were added 2. 0 μmol/L sanguinarine,2 μg/mL paclitaxel,and 2. 0μmol/L sanguinarine combined 2 μg/mL paclitaxel respectively. The morphology of A2780/Taxol cells was observed by inverted microscope. The cell proliferation were detected by MTT. The cell cycle was detected by flow cytometry. And the expressions of Caspase-3,TGF-β_1,and Smad7 proteins were detected by Western blot.Results The cells in control group were adhered to the wall and the bounday was clear. the cell volume decreased and the number of adherent cells decreased in sanguinarine group. The cells in Taxol group fall off were circularly. These changes were most obvious in the combined group. Compared with the control group,the proliferation of A2780/Taxol cells could be significantly inhibited,the expressions of Caspase-3 and Smad7 protein were up-regulated,the expression of TGF-β_1 protein was down-regulated in the rest 3 groups. The most effect was obviously seen in the combined group(P < 0. 05). Sanguinarine showed no effect on cell cycle. The cell numbers of G0/G1 and S phase decreased and the cell number of G2/M phase increased in the paclitaxel group and the combined group. Conclusion Sanguinarine could inhibit the growth of A2780/Tax-ol cells and enhance the sensitivity of paclitaxel by suppressing TGF-β_1/Smad pathway and promoting Caspase-3 expression.
引文
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[2]王新宇,谢幸.妇科恶性肿瘤的非手术治疗[J].中国牢实用妇科与产科杂,2010,26(3):163-165.
[3]梁宏.基于生物信息学挖掘卵巢癌顺铂耐药机制及潜在治疗药物[D].昆明:昆明医科大学,2016.
[4]Kalogris C,Garulli C,Pietrella L,et al.Sanguinarine suppresses basal-like breast cancer growth through dihydrofolate reductase inhibition[J].Biochem Pharmacol,2014,90(3):226-234.
[5]Gatti L,Cossa G,Tinelli S,et al.Improved apoptotic cell death in drug-resistant non-small-cell lung cancer cells by tumor necrosis factor-related apoptosis-inducing ligandbased treatment[J].J Pharmacol Exp Ther,2014,348(3):360-371.
[6]Eid SY,El-Readi MZ,Eldin EE,et al.Influence of combinations of digitonin with selected phenolics,terpenoids,and alkaloids on the expression and activity of P-glycoprotein in leukaemia and colon cancer cells[J].Phytomedicine,2013,21(1):47-61.
[7]Lonn P,Moren A,Raja E,et al.Regulating the stability of TGF beta receptors and Smads[J].Cell Res,2009,19(6):21-35.
[8]Pickup M,Novitskiy S,Moses HL.The roles of tgf beta in the tumor microenvironment[J].Nat Rev Cancer,2013,13(11):788-799.
[9]高向朋,刘清,刘涛,等.TGF-β1/Smad7信号通路在食管鳞癌上皮间质转化中的作用[J].中山大学学报(医学科学版),2015,36(1):78-87.
[10]Yi EY,Park SY,Jung SY,et al.Mitochondrial dysfunction induces EMT through the TGF-β/Smad/Snail signaling pathway in Hep3B hepatocellular carcinoma cells[J].Int J Oncol,2015,47(5):1845-1853.
[11]刘先芳,梁敬钰,孙建博.紫杉醇:具有里程碑意义的天然抗癌药物[J].世界科学技术-中医药现代化,2017,19(6):941-949.