硫利达嗪诱导SW480细胞凋亡的机制
发布时间：2018-12-19 12:29
【摘要】：目的研究硫利达嗪对结肠癌SW480细胞增殖及凋亡的影响。方法应用5~30μmol/L硫利达嗪处理SW480细胞,MTT法检测细胞增殖抑制率;Hoechst33342细胞核染色法观察细胞凋亡的形态学改变;流式细胞仪检测细胞凋亡率、细胞周期;RTq PCR分析PDCD4、c-MYC、BCL2、CCND1、CASPASE3、PARP1、CDK4、EIF4A基因表达水平;Western blotting检测AKT、pAKT、PDCD4蛋白表达水平。结果 MTT结果表明硫利达嗪抑制SW480细胞的增殖,硫利达嗪处理细胞后出现核固缩、染色质凝集和核碎片化等典型的细胞凋亡特征;流式检测表明硫利达嗪诱导G0/G1期阻滞,细胞凋亡增加。RT-PCR结果表明硫利达嗪处理细胞后PDCD4表达上调,CCND1、CDK4、c-MYC、BCL2、CASPASE3、PARP1和EIF4A表达下调。免疫印迹分析结果显示PDCD4蛋白表达上调,p-AKT蛋白表达下调。结论硫利达嗪能够抑制人结肠癌SW480细胞的增殖,并诱导其凋亡,其机制可能与抑制PI3K/AKT信号通路导致PDCD4表达水平升高有关。
[Abstract]:Objective to study the effects of talidazide on the proliferation and apoptosis of colon cancer SW480 cells. Methods SW480 cells were treated with 5 ~ 30 渭 mol/L tiridamine, the proliferation inhibition rate was detected by MTT assay, the morphological changes of apoptosis were observed by Hoechst33342 cell nuclear staining, the apoptosis rate and cell cycle were detected by flow cytometry. RTq PCR analysis of PDCD4,c-MYC,BCL2,CCND1,CASPASE3,PARP1,CDK4,EIF4A gene expression level; Western blotting detection of AKT,pAKT,PDCD4 protein expression level. Results the results of MTT showed that thiridamine inhibited the proliferation of SW480 cells, and the typical apoptotic characteristics such as nuclear pyknosis, chromatin agglutination and nuclear fragmentation appeared after treatment with thiridamine. Flow cytometry showed that tiridamine induced G0/G1 phase arrest and increased cell apoptosis. RT-PCR results showed that the expression of PDCD4 was up-regulated and the expression of CCND1,CDK4,c-MYC,BCL2,CASPASE3,PARP1 and EIF4A was down-regulated after treatment with thiridamine. Western blot analysis showed that the expression of PDCD4 protein was up-regulated and the expression of p-AKT protein was down-regulated. Conclusion tiridazide can inhibit the proliferation and induce apoptosis of human colon cancer SW480 cells. The mechanism may be related to the increase of PDCD4 expression caused by inhibition of PI3K/AKT signaling pathway.
【作者单位】： 南方医科大学中医药学院分子生物学实验室;南方医科大学肿瘤研究所;
【基金】：国家自然科学基金(81372895) 广东省自然科学基金(S2013010014275)~~
【分类号】：R96
[Abstract]:Objective to study the effects of talidazide on the proliferation and apoptosis of colon cancer SW480 cells. Methods SW480 cells were treated with 5 ~ 30 渭 mol/L tiridamine, the proliferation inhibition rate was detected by MTT assay, the morphological changes of apoptosis were observed by Hoechst33342 cell nuclear staining, the apoptosis rate and cell cycle were detected by flow cytometry. RTq PCR analysis of PDCD4,c-MYC,BCL2,CCND1,CASPASE3,PARP1,CDK4,EIF4A gene expression level; Western blotting detection of AKT,pAKT,PDCD4 protein expression level. Results the results of MTT showed that thiridamine inhibited the proliferation of SW480 cells, and the typical apoptotic characteristics such as nuclear pyknosis, chromatin agglutination and nuclear fragmentation appeared after treatment with thiridamine. Flow cytometry showed that tiridamine induced G0/G1 phase arrest and increased cell apoptosis. RT-PCR results showed that the expression of PDCD4 was up-regulated and the expression of CCND1,CDK4,c-MYC,BCL2,CASPASE3,PARP1 and EIF4A was down-regulated after treatment with thiridamine. Western blot analysis showed that the expression of PDCD4 protein was up-regulated and the expression of p-AKT protein was down-regulated. Conclusion tiridazide can inhibit the proliferation and induce apoptosis of human colon cancer SW480 cells. The mechanism may be related to the increase of PDCD4 expression caused by inhibition of PI3K/AKT signaling pathway.
【作者单位】： 南方医科大学中医药学院分子生物学实验室;南方医科大学肿瘤研究所;
【基金】：国家自然科学基金(81372895) 广东省自然科学基金(S2013010014275)~~
【分类号】：R96
【共引文献】
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