干扰 ID H-2基因对小细胞肺癌生长抑制作用的研究

摘要

[ABSTRACT]AIM:Toinvestigatetheeffectofsilencingisocitratedehydrogenase2(IDH-2)genebysmallinter-fering RNA (siRNA) on the biological characteristics of human small cell lung cancer cell line NCI -H446.METHODS:IDH-2 expression was knocked down in human small cell lung cancer cell line NCI -H446 by siRNA-IDH-2.The expression level of IDH-2 was determined by real-time PCR and Western blotting .The cell proliferation was measured by CCK-8 as-say , the protein expression of MAPK p 42 was detected by Western blotting , and the cell cycle was analyzed by flow cytome-try.The migration was observed using Transwell cell migration system .BALB/c nude mice were subcutaneously injected on the back with NCI-H446 cells transfected with siRNA-IDH-2/negative control siRNA or non-transfected cells to study the tumor growth .RESULTS:siRNA-IDH-2 remarkably down-regulated the expression of IDH-2 and MAPK p42 in the NCI-H446 cells.siRNA-IDH-2 inhibited both the proliferation and migration abilities of NCI-H446 cells, and the cell cycle was arrested in S phase as compared with negative control group .Additionally, the volume of xenograft tumors in siRNA-IDH-2 group was significantly decreased as compared with control group .CONCLUSION:siRNA-IDH-2 down-regulates the expres-sion of IDH-2 in NCI-H446 cells, reduces the cell migration efficiency and inhibits the tumor growth in vitro and in vivo.%目的：研究siRNA特异干扰沉默异柠檬酸脱氢酶2（IDH-2）基因后对人小细胞肺癌细胞NCI-H446生长作用的影响。方法：用siRNA沉默NCI-H446细胞的IDH-2基因表达，用real-time PCR及Western blotting技术检测mRNA-IDH-2和蛋白的表达；CCK-8法测定细胞生长抑制，Western blotting 检测MAPK p42的表达和流式细胞术检测细胞周期；用Transwell细胞迁移系统检测siRNA-IDH-2对NCI-H446细胞迁移能力的影响；将转染siRNA-IDH-2、阴性对照siRNA的细胞或未转染细胞皮下接种于 BALB/c 裸鼠背部，观察移植瘤的生长状况。结果：siRNA-IDH-2有效沉默NCI-H446细胞中IDH-2的表达；与对照组相比，下调IDH-2基因表达抑制NCI-H446细胞的增殖；MAPK p42蛋白表达下降且S期细胞明显增加；下调IDH-2基因显著抑制了NCI-H446细胞的迁移能力；体内实验显示siRNA-IDH-2组的肿瘤体积明显小于对照组。结论： siRNA-IDH-2能显著抑制IDH-2基因在人小细胞肺癌细胞NCI-H446中的表达，并降低细胞迁移效率，在体外和体内明显抑制NCI-H446细胞的生长。