人舌鳞癌细胞SOCS1基因沉默对细胞增殖和化疗药物敏感性的影响

目的:在人舌鳞癌细胞中观察细胞因子信号转导抑制因子1(SOCS1)沉默对细胞增殖及化疗药物敏感性的影响,探讨SOCS1作为舌鳞癌治疗靶点的研究。方法:western blot、PCR及定量PCR验证SOCS1干扰序列沉默人舌鳞癌细胞系CAL27中SOCS1的表达; MTT法检测舌鳞癌细胞对化疗药物敏感性的变化;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:将SOCS1干扰序列转染CAL27细胞后,SOCS1 mRNA及蛋白表达水平均明显下降。沉默SOCS1表达后,化疗药物卡铂和紫杉醇对CAL27细胞的半数抑制浓度(IC₅₀)均显著降低(P<0.01);转染72 h后CAL27细胞数量较对照组显著减少(P<0.05);SOCS1表达抑制后CAL27细胞G0/G1期细胞比例明显升高,而S期和G2/M期细胞比例明显减小,与对照组比较差异均有统计学意义(P<0.05)。结论:SOCS1表达抑制后,人舌鳞癌细胞株CAL27的增殖能力下降,并且对化疗药物的敏感性增强。

Effects of Suppressor of Cytokine signaling-1 Silencing on the Proliferation and Chemotherapeutic Drugs Sensitivity of Human Squamous Cell Carcinoma of Tongue.

Objective: To detect the changes of cell proliferation and chemotherapeutic drugs sensitivity of human squamous cell carcinoma of tongue 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 chemotherapeutic drugs. Methods: Western blot, PCR and real-time PCR were used to verify the down regulation of SOCS1 human squamous cell carcinoma of tongue cell (CAL27) after transfection; and the change of chemotherapeutic drugs sensitivity was detected by MTT assay; cell counting was used to measure cell proliferation rate; flow cytometry was used to detect the cell cycle. Results: Results of RT-PCR and western blot showed that the expression levels of SOCS1 mRNA and protein were significantly decreased in small hairpin SCOS1 (shSOCS1) transfected CAL27 cells. After the silence of SOCS1, the median inhibitory concentration of chemotherapeutic drugs for CAL27 cells decreased significantly (P<0.01). 72 hours after the SOCS1 silencing, the cell count of shSOCS1 cells dropped significantly compared with that of control group (P<0.05). The cell cycle arrest was promoted at the G0/G1 phase, but the percentage of cells in S phase and G2/M decreased compared to that of control groups (P<0.05). Conclusion: Inhibition of SOCS1 gene expression decreases the proliferation ability of human squamous cell carcinoma of tongue cell line CAL27, and enhances the sensitivity of CAL27 cells to chemotherapeutic drugs.