CAJ | 학술논문

目的探讨氧化还原酶基因(WWOX)对甲状腺癌细胞凋亡的影响及其作用机制.方法将携有WWOX基因的真核表达载体体外转染甲状腺乳头状癌细胞株K1(重组质粒组),筛选稳定转染的细胞并扩增培养,以转染空载质粒(空质粒组)及未经转染(空白对照组)的K1细胞作为对照.合成针对WWOX的靶向小干扰RNA(WWOX siRNA组)转染甲状腺乳头状癌细胞株TPC-1,以空白细胞(空白对照组)和转染脂质体试剂(脂质体对照组)作为对照组.采用实时定量反转录-聚合酶链反应(RT-qPCR)检测转染前后WWOX基因mRNA水平.采用Western blot法分析WWOX基因对甲状腺乳头状癌凋亡相关蛋白的影响.结果重组质粒组细胞中WWOX mRNA能稳定表达,是空质粒组及空白对照组细胞WWOX mRNA表达的4.5倍(P＜0.05).WWOX siRNA组WWOX mRNA表达明显下降,是对照组的0.2倍(P＜0.05).重组质粒组较空质粒组及空白对照组凋亡相关蛋白表达明显增加,差异有统计学意义(P＜0.05).相反,WWOX siRNA组与对照组比较,细胞凋亡相关蛋白表达明显下降(P＜0.05).结论 WWOX基因能促进甲状腺癌细胞的凋亡,其机制可能与激活线粒体凋亡信号通路有关.
목적 탐토양화환원매기인(WWOX)대갑상선암세포조망적영향급기작용궤제.방법 장휴유WWOX기인적진핵표체재체체외전염갑상선유두상암세포주K1(중조질립조),사선은정전염적세포병확증배양,이전염공재질립(공질립조)급미경전염(공백대조조)적K1세포작위대조.합성침대WWOX적파향소간우RNA(WWOX siRNA조)전염갑상선유두상암세포주TPC-1,이공백세포(공백대조조)화전염지질체시제(지질체대조조)작위대조조.채용실시정량반전록-취합매련반응(RT-qPCR)검측전염전후WWOX기인mRNA수평.채용Western blot법분석WWOX기인대갑상선유두상암조망상관단백적영향.결과 중조질립조세포중WWOX mRNA능은정표체,시공질립조급공백대조조세포WWOX mRNA표체적4.5배(P＜0.05).WWOX siRNA조WWOX mRNA표체명현하강,시대조조적0.2배(P＜0.05).중조질립조교공질립조급공백대조조조망상관단백표체명현증가,차이유통계학의의(P＜0.05).상반,WWOX siRNA조여대조조비교,세포조망상관단백표체명현하강(P＜0.05).결론 WWOX기인능촉진갑상선암세포적조망,기궤제가능여격활선립체조망신호통로유관.
Objective To investigate the effects of WW domail-containing oxidoreductase (WWOX)-mediated apoptosis on papillary thyroid carcinoma in vitro and its possible mechanism.Methods A eukaryotic expression vector containing WWOX was transfected into papillary thyroid cancer cell line K1 (recombinant plasmid group),and positive cell clones were selected and amplified.Synthesized small interfering RNA (siRNA) targeting WWOX (WWOX siRNA group) was transfected into TPC-1 cells.On the other hand,TPC-1 cells transfected with Lipofectamine 2000 (liposomes control group) and vacant TPC-1 cells (blank control group) were used as controls.The expression of WWOX mRNA was detected by real-time quantitative reverse transcriptase-polymerase chain reaction (RT-qPCR).The effects of WWOX on apoptosis releated protein were analyzed by Western blotting.Results Recombinant plasmid group cells could steadily express WWOX mRNA,which was more than 4.5 times higher than plasmid group and blank control group (P ＜ 0.05).The absorbance of TPC-1 cells decreased greatly after transfection with WWOX siRNA,which was 0.2 times lower than in control groups (P ＜ 0.05).Forced expression of WWOX in K1 cells enhanced the expression of apoptosis releated protein.Conversely,knockdown of WWOX in TPC-1 cells using siRNA targeting WWOX decreased the expression of apoptosis releated protein.Conclusion WWOX can promote apoptosis of human PTC cells through a mechanism which may be assocoated with activation of the mitochondrial apoptosis pathway.