缺氧诱导因子-1α、生长素基因沉默对CNE-2鼻咽癌细胞放疗及凋亡敏感性的影响  

Effects of HIF-1α and Survivin gene silencing on radiotherapy and apoptosis sensitivity of CNE-2 nasopharyngeal carcinoma cells

作　　者：徐茂林 陈晓华 孙波 XU Mao-Lin;CHEN Xiao-Hua;SUN Bo(Department of Otolaryngology,First Hospital of Zibo City,Zibo 255200,China)

机构地区：[1]淄博市第一医院耳鼻咽喉科

出　　处：《中国免疫学杂志》2019年第18期2249-2252,2257,共5页Chinese Journal of Immunology

摘　　要：目的:探讨缺氧诱导因子-1α(HIF-1α)、生长素(Survivin)基因沉默对CNE-2鼻咽癌细胞放疗及凋亡敏感性的影响。方法:将合成的靶向沉默HIF-1α和(或)Survivin的microRNA质粒转染到鼻咽癌CNE-2细胞,接种至BALB/c裸鼠右侧腋下,根据接种的转染质粒不同,将裸鼠分为阴性对照组、空载组、联合干扰组、HIF-1α干扰组、Survivin干扰组。成瘤后每3d放射治疗1次,每次5只,给予5GyX-放射治疗,共15GyX。放射结束后3d,处死裸鼠取瘤体计算体积。分别采用RT-qPCR、Westernblot法检测组织中HIF-1α、Survivin的mRNA及蛋白表达水平,采用TUNEL法检测细胞凋亡。结果:①各组CNE-2细胞均能在裸鼠体内良好成瘤,成瘤率100%(35/35)。肿瘤生长曲线显示,随着成瘤时间的延长,空载组、阴性对照组裸鼠的肿瘤生长速度明显更快,肿瘤体积显著更大,其他三组的体积变化率显著较低(P<0.05)。②阴性对照组与空载组的HIF-1α、Survivin mRNA表达量、蛋白表达量无明显差异(P>0.05),但显著高于其他三组(P<0.05)。③TUNEL检测结果显示,正常细胞胞核蓝染,形态均一,无黄染颗粒。凋亡细胞胞核固缩,形态异常,出现棕黄色颗粒。HIF-1α干扰组、Survivin干扰组、联合干扰组之间,两两比较,凋亡细胞数量无明显差异(P>0.05)。但三组的凋亡细胞数量均明显多于空载组、阴性对照组(P<0.05)。结论:单独的HIF-1α、Survivin基因干扰能有效增强鼻咽癌移植瘤裸鼠模型对放疗的敏感性,但联合干扰后敏感性未明显增强。Objective:To investigate the effect of HIF-1 and Survivin gene silencing on the radiotherapy and apoptosis sensitivity of CNE-2 nasopharyngeal carcinoma cells.Methods:the synthesized plasmid microRNA,which silenced HIF-1αand/or Survivin,was transfected into CNE-2 cells and inoculated into BALB/c nude mice.Radiation therapy was given after tumor formation.After radiation,the nude mice were killed and the volume was calculated.The levels of mRNA and protein in HIF-la and Survivin were detected,and apoptosis was detected by TUNEL.Results:①With the prolongation of the tumor formation time,the tumor growth rate of the nude mice in the unloaded group and the negative control group was significantly faster and the volume of the tumor was significantly larger.The volume change rate of the HIF-1 alpha interference group,the Survivin interference group and the combined interference group was significantly lower(P<0.05).②There was no significant difference in the expression of HIF-1αand Survivin mRNA(P>0.05)in the negative control group and the no-load group(P>0.05),but significantly higher than that in the other three groups(P<0.05).③The results of TUNEL detection showed that there was no significant difference in the number of apoptotic cells between HIF-1αinterference group,Survivin interference group and combined interference group(P>0.05).However,the number of apoptotic cells in the empty group and negative control group were significantly less than those in the other three groups(P<0.05).Conclusion:The interference of HIF-1αand Survivin gene alone can effectively enhance the sensitivity of nude mice model to radiotherapy in nasopharyngeal carcinoma,but the sensitivity is not significantly enhanced after combined interference.

关 键 词：缺氧诱导因子-1Α 生长素 鼻咽癌 

分 类 号：R739.62[医药卫生—肿瘤]