金属硫蛋白-3表达对A549肺癌株生物学功能的影响  

Effects of metallothionein-3 expression on biological behaviors of A549 lung cancer cells

作　　者：陈重[1] 魏立[1] 务森[1] 张宁[1] 朱晓明[1] 陈明耀[1] 

机构地区：[1]河南省人民医院胸外科,郑州450003

出　　处：《中华实验外科杂志》2014年第10期2200-2203,共4页Chinese Journal of Experimental Surgery

摘　　要：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.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.

关 键 词：肺癌 金属硫蛋白-3 脱噬作用 

分 类 号：R734.2[医药卫生—肿瘤]