CAJ | 학술논문

目的：研究线粒体动力学蛋白Mfn2和Drp1在甲状腺鳞癌SW579细胞中的表达和线粒体分裂抑制剂-1（Mdivi-1）对SW579细胞增殖、凋亡及侵袭能力的影响。方法采用RT-PCR检测Mfn2和Drp1 mRNA，Western blot检测Mfn2和Drp1蛋白在Nthy-ori 3-1和SW579两种细胞中的表达；然后将SW579分为对照组（DMSO,0.1%）和Mdivi-1低、中、高剂量组（浓度分别为15、30、45μmol/L），培养16 h，采用MTT法检测细胞增殖能力的变化，用荧光分光光度计检测线粒体膜电位的变化，用RT-PCR检测细胞色素C（Cyt C）和Caspase-3 mRNA的变化，Western blot检测Cyt C和Caspase-3蛋白的变化，用Transwell小室侵袭实验检测侵袭能力的变化。结果与Nthy-ori 3-1细胞系相比，SW579细胞系中Mfn2 mRNA和蛋白表达明显降低，Drp1 mRNA和蛋白表达明显升高（P＜0.01）；与对照组相比，Mdivi-1各组SW579细胞存活率和线粒体膜电位均明显降低，Cyt C和Caspase-3的mRNA及蛋白表达均明显增加，侵袭能力明显减弱（均P＜0.01），并且呈药物浓度依赖性。结论甲状腺鳞癌SW579细胞中存在线粒体动力学异常，Mdivi-1可以抑制此细胞的增殖和侵袭，并能诱导其凋亡。
목적：연구선립체동역학단백Mfn2화Drp1재갑상선린암SW579세포중적표체화선립체분렬억제제-1（Mdivi-1）대SW579세포증식、조망급침습능력적영향。방법채용RT-PCR검측Mfn2화Drp1 mRNA，Western blot검측Mfn2화Drp1단백재Nthy-ori 3-1화SW579량충세포중적표체；연후장SW579분위대조조（DMSO,0.1%）화Mdivi-1저、중、고제량조（농도분별위15、30、45μmol/L），배양16 h，채용MTT법검측세포증식능력적변화，용형광분광광도계검측선립체막전위적변화，용RT-PCR검측세포색소C（Cyt C）화Caspase-3 mRNA적변화，Western blot검측Cyt C화Caspase-3단백적변화，용Transwell소실침습실험검측침습능력적변화。결과여Nthy-ori 3-1세포계상비，SW579세포계중Mfn2 mRNA화단백표체명현강저，Drp1 mRNA화단백표체명현승고（P＜0.01）；여대조조상비，Mdivi-1각조SW579세포존활솔화선립체막전위균명현강저，Cyt C화Caspase-3적mRNA급단백표체균명현증가，침습능력명현감약（균P＜0.01），병차정약물농도의뢰성。결론갑상선린암SW579세포중존재선립체동역학이상，Mdivi-1가이억제차세포적증식화침습，병능유도기조망。
Objective To detect the expression of mitochondrial dynamics proteins (Mfn2 and Drp1) in thyroid squa?mous carcinoma cell line SW579 and the effects of Mitochondrial division inhibitor, Mdivi-1, on proliferation, apoptosis and invasion of SW579. Methods In SW579 and Nthy-ori 3-1 cell lines, the expression levels of Mfn2 and Drp1 were deter?mined by western blot while the transcription level of Mfn2 and Drp1 mRNA were measured by RT-PCR. Then, SW579 cells were divided into control group (DMSO, 0.1%) and Mdivi-1 low, medium and high dose groups (Mdivi-1 of 15,30 and 45μmol/L were incubated with cells for 16 hours respectively). Then the ability of cell proliferation was detected using MTT assay, the mitochondrial membrane potential was determined by fluorescence spectrophotometer, the expression levels of cy?tochrome C and Caspase-3 were quantified by Western blot and the transcription level of the Cyt C and Caspase-3 mRNA were determined by RT-PCR. The ability of invasion in each group was measured with Transwell assays. Results Com?pared with Nthy-ori 3-1, the mRNA transcription and protein expression levels of the Mfn2 was remarkably decreased, while the mRNA transcription and protein expression of the Drp1 was significantly increased in SW579 cells (P<0.01). Compared with control group, the cell survival rates and mitochondrial membrane potential of SW579 were decreased dramat?ically (P<0.01). The mRNA transcription and protein expression of the cytochrome C and Caspase-3 were increased dra?matically (P<0.01) and the capability of invasion was markedly decreased in all the Mdivi-1 groups in a dosage dependent manner compared with those in control groups (P<0.01). Conclusion Abnormal mitochondrial dynamics may be involved in thyroid squamous cell carcinoma SW579 cells;Mdivi-1 can inhibit the cell proliferation and invasion as well as induce apoptosis.