CAJ | 학술논문

目的观察藤黄酸对人肺腺癌细胞株A549增殖和凋亡的影响,及其对甾体类受体共激活因子3(SRC-3)表达的调控作用.方法以不同浓度的藤黄酸处理人肺腺癌A549细胞,采用四甲基偶氮唑蓝(MTT)法检测细胞的增殖活性,Hoechst 33258染色法分析细胞凋亡的改变,共聚焦显微镜观察A549细胞中SRC-3蛋白的分布情况,Western blot和逆转录聚合酶链反应(RT-PCR)法检测藤黄酸对A549细胞中SRC-3 mRNA和蛋白表达的影响.结果藤黄酸能明显抑制A549细胞的增殖,其抑制作用呈时间和浓度依赖性,24和48 h的半数抑制浓度(IC_(50))分别为(3.17.4±0.13)μmol/L和(1.85±0.08)gmol/L.藤黄酸能使A549细胞出现典型的细胞凋亡形态改变.空白对照组的A549细胞中,SRC-3 mRNA和蛋白均呈高表达,且主要分布于细胞核;经藤黄酸处理后,A549细胞中SRC-3 mRNA和蛋白的表达水平均明显下降.结论藤黄酸能明显抑制人肺腺癌细胞3.549的增殖,并可能通过下调SRC-3 mRNA和蛋白的表达量而诱导A549细胞产生凋亡.藤黄酸有望成为治疗肺癌的新型靶点药物.
목적 관찰등황산대인폐선암세포주A549증식화조망적영향,급기대치체류수체공격활인자3(SRC-3)표체적조공작용.방법 이불동농도적등황산처리인폐선암A549세포,채용사갑기우담서람(MTT)법검측세포적증식활성,Hoechst 33258염색법분석세포조망적개변,공취초현미경관찰A549세포중SRC-3단백적분포정황,Western blot화역전록취합매련반응(RT-PCR)법검측등황산대A549세포중SRC-3 mRNA화단백표체적영향.결과 등황산능명현억제A549세포적증식,기억제작용정시간화농도의뢰성,24화48 h적반수억제농도(IC_(50))분별위(3.17.4±0.13)μmol/L화(1.85±0.08)gmol/L.등황산능사A549세포출현전형적세포조망형태개변.공백대조조적A549세포중,SRC-3 mRNA화단백균정고표체,차주요분포우세포핵;경등황산처리후,A549세포중SRC-3 mRNA화단백적표체수평균명현하강.결론 등황산능명현억제인폐선암세포3.549적증식,병가능통과하조SRC-3 mRNA화단백적표체량이유도A549세포산생조망.등황산유망성위치료폐암적신형파점약물.
Objective To investigate the effects of gambogic acid(GA)on the proliferation inhibition and apoptosis induction in Human lung adenocarcinoma A549 cells in vitro,as well as the regulation of steroid receptor coactivator-3(SRC-3)to explore the relationship between them.Methods The effect of GA on the growth of A549 cells wag studied by MTr assay.Apoptosis waa detected by Hoechst 33258staining.The localization of SRC-3 was determined by confocal laser scanning microscopy.Western blot and RT-PCR technique were applied to assess the expression of SRC-3.Results GA presented a striking pmliferation inhibition potency on A549 cells in vitto.as well as apeptosis induction activity in a time-and dose-dependent manner.The IC_(50) value for 24 h was(3.17 4±0.13)μmol/L.Overexpression of SRC-3 was found in A549 cells.wheras the SRC-3 protein and mRNA expression levels were significantly downregulated in A549 cells induced by GA in a dose-dependent manner.The location of SRC-3 was situated mainly in the cell nuclei.Conclusion GA exhibits a potent proliferation inhibition and apoptosis induction in human lung adeoocarcinoma A549 cells,which might correspond to the downregulation of the expression of SRC-3.Thus it promises to be a new target drug for lung cancer treatment.