细胞因子信号转导抑制因子1沉默对人恶性黑色素瘤细胞增殖和干扰素-γ敏感性的影响  

Effects of suppressor of cytokme signaling-1 silencing on the proliferation and interferon gamma susceptibility of human malignant melanoma cell

作　　者：涂志全 杭华 段力[1] 温宏升[1] 

机构地区：[1]上海市第八人民医院肿瘤科,上海200235 [2]上海市徐汇区漕河泾卫生服务中心,上海200235

出　　处：《现代肿瘤医学》2016年第3期367-371,共5页Journal of Modern Oncology

摘　　要：目的:探讨沉默细胞因子信号转导抑制因子1(SOCS1)对人恶性黑色素瘤细胞增殖和干扰素-γ(IFN-γ)敏感性的影响。方法:采用Western blot和RT-PCR检测SOCS1干扰序列沉默人恶性黑色素瘤细胞系Me1526中SOCS1的表达;然后给予IFN-γ刺激后,观察信号传导及转录激活因子(STAT)1及磷酸化STAT(pSTAT)1和IFN-γ调节因子1(IRF-1)表达水平变化。MTT法检测恶性黑色素瘤细胞对IFN-γ敏感性的变化。细胞计数的方法观察细胞的增殖速度。采用流式细胞术的方法检测细胞周期的变化。结果:SOCS1干扰序列可有效沉默Me1526中SOCS1的表达,并且细胞周期中S期明显延长,抑制SOCS1的表达后,增殖实验表明Me1526细胞增殖能力下降;给予IFN-γ刺激后pSTAT1及IRF-1的表达水平显著升高,MTT实验表明SOCS1表达抑制后IFN-γ对Me1526细胞的半数抑制浓度(IC_(50))显著降低。结论:SOCS1表达抑制后,人恶性黑色素瘤细胞株Me1526的增殖能力下降,并且其对IFN-γ的敏感性增强。Objective：To study the changes of biological behaviors and IFN-γ susceptibility of human melanoma cells after suppressor of cytokine signaling-1（SOCS1） gene silencing,and to explore the SOCS1 as the target of anti-tumor therapy through enhancing the function of IFN-γ.Methods：Western blot,RT-PCR were used to verify the downregulation of SOCS1 in mouse erythroleukemia cell（Mel）526 after transfection.Subsequently,Mel526 was stimulated with IFN-γ.Western blot was used to detect the expression of signal transducers and activators of transcription（STAT）1 and phosphorylation STAT（pSTAT）1;and the change of IFN-γ susceptibility was detected by MTT assay.RT-PCR was used to detect mRNA of interferon regulatory factor-1（IRF-1）.Flow cytometry was used to detect the cell cycle.Results：SOCS1 can be efficiently down regulated after modulation.Silencing SOCS1 in Mel526 could significantly enhance the expression of pSTAT1 and IRF-1 following the stimulation with IFN-γ.Meanwhile,the S phase in cell cycle was prolonged after the SOCS1 silencing.After SOCS1 gene silencing,the proliferation ability of Mel526 cells were decreased,and the median inhibitory concentration of IFN-γ for Mel526 cells also decreased significantly.Conclusion：Inhibition of SOCS1 gene expression decreases the proliferation ability of human melanoma cell line Mel526,and enhances the sensitivity of Mel526 cells to IFN-γ.

关 键 词：黑色素瘤 SOCS1 肿瘤免疫 干扰素-Γ 

分 类 号：R730.51[医药卫生—肿瘤]