热疗联合Id-1基因沉默对舌鳞癌CAL-27细胞增殖及侵袭的影响

目的: 探讨热疗联合Id-1基因沉默对舌鳞癌细胞增殖及侵袭的影响及其机制。方法: 将培养的舌鳞癌细胞株CAL-27分为4组。①沉默Id-1组（Id-1-siRNA）—用siRNA基因转染法沉默舌鳞癌CAL-27细胞中Id-1的表达,RT-PCR法检测其Id-1的mRNA表达变化;②热疗组（HT）—细胞置于42.5℃恒温培养箱内40min;③联合组（HT+ Id-1-siRNA）—siRNA基因转染法沉默细胞内Id-1的表达后进行42.5℃、40 min的热疗处理;④空白对照组—常规培养细胞。通过CCK8和Transwell侵袭实验,分别检测CAL-27细胞的增殖能力和侵袭能力变化; Western免疫印迹法检测CAL-27细胞中PI3K、磷酸化Akt（p-Akt）蛋白表达变化。采用SPSS19.0软件包对数据进行统计学分析。结果: siRNA能成功沉默CAL-27细胞内Id-1的表达,沉默Id-1组与空白对照组相比,Id-1的mRNA表达水平下降81.6%。沉默Id-1和热疗均可抑制CAL-27细胞的增殖和侵袭能力,下调PI3K、p-Akt蛋白表达（P<0.05）,热疗联合沉默Id-1作用于CAL-27细胞后的结果更为显著（P<0.01）。结论: 热疗协同沉默Id-1可显著抑制人舌鳞癌细胞的增殖及侵袭能力,其作用是通过PI3K/Akt信号通路来实现。

Effect of hyperthermia combined with silencing Id-1 gene on proliferation and invasion of tongue squamous cell carcinoma cell line CAL-27

PURPOSE: To investigate the effects and regulatory mechanisms of hyperthermia combined with silencing Id-1 gene on proliferation and invasion of tongue squamous cell carcinoma in vitro. METHODS: Cal-27 cells were divided into 4 groups. In silencing Id-1 group (Id-1-siRNA), siRNA was used to silence the expression of Id-1 in the CAL-27 cells, and the mRNA expression of Id-1 was detected by RT-PCR. In hyperthermia group (HT), CAL-27 cells were placed into constant temperature incubator at 42.5℃ for 40 minutes. In combined group (HT+Id-1-siRNA), hyperthermia combined with silencing Id-1 gene were exerted on CAL-27 cells. Normally cultured CAL-27 cells were used as control group. The proliferation and invasion ability were analyzed by CCK8 assay and Transwell chamber test,and the protein expression of PI3K, p-Akt was detected by Western blotting. SPSS 19.0 software package was used to analyze the data. RESULTS: The expression level of Id-1 mRNA decreased by 81.6% in the Id-1-siRNA group, compared with the blank control group. Both silencing Id-1 and hyperthermia not only inhibited the proliferation and invasion of CAL-27 cells（P<0.05）, but also down-regulated the protein expression of PI3K, p-Akt; however, the combination of hyperthermia and Id-1-siRNA contributed to more significant results. CONCLUSIONS: Hyperthermia in conjunction with silencing Id-1 could reduce the proliferation and invasion of CAL-27 cell line of tongue squamous cell carcinoma by down-regulating PI3K/Akt signaling pathway.