CAJ | 학술논문

目的探讨新近克隆出来的6次跨膜的前列腺上皮抗原--STEAP1基因的功能.方法从人前列腺癌组织钓取STEAP1基因,亚克隆人表达载体.稳定转染至甲状腺上皮细胞(FRTAP2320).利用RT-PCR和Western印迹方法鉴定STEAP1基因和蛋白表达.采用细胞生长曲线观察细胞生长情况,二氯荧光黄双乙酸盐测定细胞内活性氧(ROS)水平.结果成功获得了稳定表达STEAP1基因的甲状腺上皮细胞系,该细胞系生长速度明显高于对照组;转染STEAP1基因的甲状腺上皮细胞内ROS水平高于对照组(P＜0.01).结论 STEAP1的高表达通过诱导细胞内ROS水平增高,促进细胞快速生长.
목적 탐토신근극륭출래적6차과막적전렬선상피항원--STEAP1기인적공능.방법 종인전렬선암조직조취STEAP1기인,아극륭인표체재체.은정전염지갑상선상피세포(FRTAP2320).이용RT-PCR화Western인적방법감정STEAP1기인화단백표체.채용세포생장곡선관찰세포생장정황,이록형광황쌍을산염측정세포내활성양(ROS)수평.결과 성공획득료은정표체STEAP1기인적갑상선상피세포계,해세포계생장속도명현고우대조조;전염STEAP1기인적갑상선상피세포내ROS수평고우대조조(P＜0.01).결론 STEAP1적고표체통과유도세포내ROS수평증고,촉진세포쾌속생장.
Objective To investigate the STEAP1 gene function of the newly discovered gene six transmembrAne epithelial antigen of the prostate-1(STEAP1).Methods Total RNA was obtained from human prostate cancer tissue and underwent PCR amplification.The full length of STEAP1 gene thus obtained was cloned. Mammalian expression vector pcDNA3. 1-STEAP1 was constructed and Stably transfected into the human thyroid epithelial cells of the line FRTAP2320.A growth curve of the transfected cells was drawn.The intracellular reactive oxygen species(ROS)level was determined by flow cytometry (FC)with dichlorodihydrofluorescein diacetate(DCFHDA).Results The growth curve showed that the STEAP1 transfected cells grew faster than the control cells.FC showed that the fluorescence intensity of he intracellular ROS of the STEAP1 transfected cells was 42.13±1.13,significantly higher than those of the cell transfected with blank plasmid(10.02±1.42)and un-transfected cells(13.02 ±2.42,both P＜0.01).Conclusion STEAP1 promotes the cell growth through inducing the intracellular ROS level.