摘要
背景与目的:蛋白酶体抑制剂是一种新型抗肿瘤靶向治疗药物,其作用机制复杂。本研究前期的工作已经证实,蛋白酶体抑制剂可以有效抑制人喉鳞癌Hep-2细胞的增殖并诱导其凋亡,但它同时也诱导了p-STAT3蛋白表达水平的升高。本研究旨在探讨pshSTAT3抑制p-STAT3蛋白表达能否增强蛋白酶体抑制剂MG-132对喉癌细胞的抗肿瘤作用。方法:以Hep-2细胞为实验对象,利用四甲基偶氮唑蓝(MTT)法、流式细胞术(flowcytometry,FCM)检测MG-132单独及联合pshSTAT3时,细胞增殖抑制率及凋亡率的变化;Western印迹法检测各组p-STAT3蛋白表达情况。结果:MTT检测结果显示,联合组细胞的增殖抑制作用最强,与MG-132组及pshSTAT3组相比较差异均有统计学意义(P<0.01)。FCM检测结果显示,联合组细胞出现明显的凋亡峰,其凋亡率显著高于MG-132组和pshSTAT3组,差异极有统计学意义(P<0.01)。Western印迹法检测结果显示,经2.5μmol/LMG-132处理Hep-2细胞后p-STAT3蛋白表达显著增强;联合组及pshSTAT3组p-STAT3蛋白表达明显减弱。结论:pshSTAT3可以抑制MG-132诱导的p-STAT3蛋白表达,从而提高蛋白酶体抑制剂MG-132对喉癌细胞的抗肿瘤作用。
Background and purpose：Proteasome inhibitors constitute a novel class of antitumor agents that has a complex mechanism of action.Previous studies have confirmed that proteasome inhibitor MG-132 can significantly inhibit Hep-2 cell growth and induce cell apoptosis in a manner that is dependent on dosage and time.But it also induced p-STAT3 protein expression.The aim of this study was to explore whether the STAT3 gene can,by transfecting short hair pin RNA （shRNA）,enhance the anti-tumor effect of MG-132 on human laryngeal carcinoma cells.Methods：Hep-2 cells were plated into 96-well and 6-well plates and incubated overnight.Then,they were treated with MG-132 alone and combined with pshSTAT3.Their cell growth was detected by MTT assay,and apoptosis was examined with flow cytometry.The protein expression of p-STAT3 was detected by Western blotting.Results：MTT assay showed that a combined group inhibited the proliferation of Hep-2 cells compared to the MG-132 group and pshSTAT3 group （P0.01）.Flow cytometry showed that apoptosis of the combined group was significantly higher than the MG-132 group and pshSTAT3 group （P0.01）.Western blotting showed that the p-STAT3 protein expression up-regulation was observed in the MG-132 group,whereas down-regulation was expressed in the combined group and pshSTAT3 group.Conclusion：The shRNA targeting STAT3 gene can prevent the up-regulation of p-STAT3 protein following a MG-132 treatment thereby significantly enhancing the anti-tumor effect of the protease inhibitor,MG-132,on human laryngeal carcinoma cells.
出处
《中国癌症杂志》
CAS
CSCD
北大核心
2010年第3期173-177,共5页
China Oncology
基金
全军医学科学技术研究"十一五"计划面上A类项目(项目编号:06MA076)
