CA4P诱导大肠癌细胞死亡机制研究及相关通路检测

发布时间：2018-07-08 09:27

本文选题：药学 + CAP　；参考：《中国科技论文》2017年18期

【摘要】：为探究康普瑞汀磷酸二钠(CA4P)诱导大肠癌SW480细胞死亡机制及相关通路的检测,体外培养SW480细胞,分别给予不同浓度的CA4P进行刺激。采用CCK-8法测定不同浓度药物对SW480细胞活性的抑制率。采用流式细胞术(flow cytometry,FCM)测定SW480细胞凋亡率。采用显微镜观察药物作用24、48h后细胞形态。采用Western blotting检测SW480细胞自噬相关蛋白LC3B、Atg7和Beclin-1的表达情况。Path Scan对相关的激活通路进行检测。CCK-8结果显示,药物浓度50μmol/L时,体外CA4P对大肠癌细胞的抑制率呈递增趋势;FCM检测表明,CA4P作用后,细胞凋亡率无明显变化;显微镜观察在药物作用24h,48h后,SW480细胞的形态发生显著改变。Western blotting结果显示,CA4P刺激SW480使得自噬相关蛋白LC3B、Beclin-1、Atg7表达水平升高,差异具有统计学显著性意义(P0.05),phospho-p53表达上调。PathScan结果表明,细胞内信号通路Erk1/2、Akt、p53被激活,AMPKα被抑制。CA4P诱导大肠癌细胞死亡可能是通过自噬途径,并且可能与Erk1/2、Akt、p53的上调和AMPKα的下调有关。
[Abstract]:To investigate the mechanism of SW480 cell death induced by CA4P (CA4P) and the detection of the related pathway in colorectal cancer cells, SW480 cells were cultured in vitro and stimulated by different concentrations of CA4P. The inhibitory rate of different concentrations of drugs on the activity of SW480 cells was measured by CCK-8 method. SW480 cells were measured by flow cytometry (flow cytometry, FCM). Apoptosis rate. The morphology of the cells after 24,48h was observed by microscope. Western blotting was used to detect the expression of autophagy related protein LC3B, Atg7 and Beclin-1..Path Scan was used to detect the activation pathway of.Path Scan.CCK-8 results showed that when the drug concentration was 50 mu mol/L, the inhibition rate of colorectal cancer cells increased in vitro. FCM detection showed that there was no obvious change in the cell apoptosis rate after the action of CA4P, and the morphology of SW480 cells changed significantly after the effect of the drug action 24h and 48h, and the.Western blotting results showed that CA4P stimulated SW480 made the autophagy associated protein LC3B, Beclin-1, and Atg7 expression level increased, and the difference was statistically significant. The up-regulated expression of.PathScan in ho-p53 showed that the intracellular signaling pathway Erk1/2, Akt, p53 were activated, and AMPK a was inhibited by.CA4P to induce colorectal cancer cell death by autophagy pathway, and may be associated with the up regulation of Erk1/2, Akt, p53 and AMPK alpha.
【作者单位】：南方医科大学南方医院;贵州医科大学生理教研室;南方医科大学中医药学院;
【基金】：广东省自然科学基金资助项目(2014A030313167) 高等学校博士学科点专项科研基金资助项目(20134433120014,20134433110007) 国家自然科学基金资助项目(81472315)
【分类号】：R735.34

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