【摘要】：目的研究硫利達(dá)嗪對結(jié)腸癌SW480細(xì)胞增殖及凋亡的影響。方法應(yīng)用5~30μmol/L硫利達(dá)嗪處理SW480細(xì)胞,MTT法檢測細(xì)胞增殖抑制率;Hoechst33342細(xì)胞核染色法觀察細(xì)胞凋亡的形態(tài)學(xué)改變;流式細(xì)胞儀檢測細(xì)胞凋亡率、細(xì)胞周期;RTq PCR分析PDCD4、c-MYC、BCL2、CCND1、CASPASE3、PARP1、CDK4、EIF4A基因表達(dá)水平;Western blotting檢測AKT、pAKT、PDCD4蛋白表達(dá)水平。結(jié)果 MTT結(jié)果表明硫利達(dá)嗪抑制SW480細(xì)胞的增殖,硫利達(dá)嗪處理細(xì)胞后出現(xiàn)核固縮、染色質(zhì)凝集和核碎片化等典型的細(xì)胞凋亡特征;流式檢測表明硫利達(dá)嗪誘導(dǎo)G0/G1期阻滯,細(xì)胞凋亡增加。RT-PCR結(jié)果表明硫利達(dá)嗪處理細(xì)胞后PDCD4表達(dá)上調(diào),CCND1、CDK4、c-MYC、BCL2、CASPASE3、PARP1和EIF4A表達(dá)下調(diào)。免疫印跡分析結(jié)果顯示PDCD4蛋白表達(dá)上調(diào),p-AKT蛋白表達(dá)下調(diào)。結(jié)論硫利達(dá)嗪能夠抑制人結(jié)腸癌SW480細(xì)胞的增殖,并誘導(dǎo)其凋亡,其機(jī)制可能與抑制PI3K/AKT信號通路導(dǎo)致PDCD4表達(dá)水平升高有關(guān)。
[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.
【作者單位】： 南方醫(yī)科大學(xué)中醫(yī)藥學(xué)院分子生物學(xué)實(shí)驗(yàn)室;南方醫(yī)科大學(xué)腫瘤研究所;
【基金】：國家自然科學(xué)基金(81372895) 廣東省自然科學(xué)基金(S2013010014275)~~
【分類號】：R96

【共引文獻(xiàn)】

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