CAJ | 학술논문

目的观察金属硫蛋白-3(MT-3)表达对A549肺癌细胞株的增殖、凋亡和生物学功能的影响.方法应用基因转录方法检测MT-3表达对A549肺癌细胞株的抗增殖作用和生物学功能的影响;通过免疫组织化学法和逆转录-聚合酶链反应(RT-PCR)技术检测MT-3 mRNA在A549肺癌株中的表达水平,半定量RT-PCR(SqRT-PCR)测定转染后A549肺癌细胞中c-fos、c-jun mRNA水平表达变化,并通过显微镜、苏木素-伊红(HE)染色、透射电镜等观察凋亡细胞的形态学改变.结果 RT-PCR和免疫细胞化学证实转染后A549肺癌细胞中有MT-3稳定表达;SqRT-PCR证实转染后A549肺癌细胞中c-fos、c-jun mRNA水平显著降低(x2=2.536,P＜0.05).噻唑蓝(MTT)比色法表明随着MT-3与肺癌细胞效靶比增加及作用时间延长,抑制率明显增强(x2=2.795,P＜0.05).MT-3作用于肺癌细胞24 h后,形态学观察肺癌细胞发生凋亡或坏死.结论体外制备的MT-3对肺癌A549细胞具有抑制增殖和促进凋亡作用,其可能是通过负性调控信号通路降低A549肺癌细胞株中c-fos、c-jun的表达.
목적 관찰금속류단백-3(MT-3)표체대A549폐암세포주적증식、조망화생물학공능적영향.방법 응용기인전록방법검측MT-3표체대A549폐암세포주적항증식작용화생물학공능적영향;통과면역조직화학법화역전록-취합매련반응(RT-PCR)기술검측MT-3 mRNA재A549폐암주중적표체수평,반정량RT-PCR(SqRT-PCR)측정전염후A549폐암세포중c-fos、c-jun mRNA수평표체변화,병통과현미경、소목소-이홍(HE)염색、투사전경등관찰조망세포적형태학개변.결과 RT-PCR화면역세포화학증실전염후A549폐암세포중유MT-3은정표체;SqRT-PCR증실전염후A549폐암세포중c-fos、c-jun mRNA수평현저강저(x2=2.536,P＜0.05).새서람(MTT)비색법표명수착MT-3여폐암세포효파비증가급작용시간연장,억제솔명현증강(x2=2.795,P＜0.05).MT-3작용우폐암세포24 h후,형태학관찰폐암세포발생조망혹배사.결론 체외제비적MT-3대폐암A549세포구유억제증식화촉진조망작용,기가능시통과부성조공신호통로강저A549폐암세포주중c-fos、c-jun적표체.
Objective To study the efects of metallothionein-3 (MT-3) expression on biological behaviors of A549 lung cancer cells.Methods The anti-proliferation and the cytotoxicity of MT-3 on A549 cells were detected by methyl thiazol tetrazolium (MTT) assay.The MT-3 expression was observed in A549 cells,and the mRNA level was detected by reverse transcription-polymerase chain reaction (RT-PCR).The morphological changes of the apoptotic cells were observed by invert microscope,hematoxylin and eosin (HE) stain,transmission electron microscope and acridine orange/ethidium bromide (AO/EB) double fluorescent staining.The data were analyzed by using SPSS 14.0 statistical software.Results The MT-3 expression in A549 cells transfected with MT-3 gene was confirmed by RT-PCR and immunocytochemical method,respectively.Compared with the control group,-fos and c-jun mRNA levels were decreased significantly in PEFMT-3 transfected A549 cells (x2 =2.536,P ＜ 0.05).MTT assay revealed that the inhibitory rate was enhanced obviously with the increase of effect/target rate and extension of time (x2 =2.795,P ＜0.05).The apoptosis or necrosis of A549 cells was morphologically observed after lung cancer cells were induced by MT-3 for 24 h.Conclusion MT-3 could inhibitit proliferation of A549 cells and promote apoptosis of A549 cells probably by negatively regulating the signaling pathway to ruduce the expression of c-fos and c-jun.