【摘要】 目的探讨用特异性短发卡RNA(small hairpin RNA,shRNA)抑制表皮生长因子受体(epidermal growth factor receptor,EGFR)表达对鳞状细胞癌Colo-16细胞凋亡的影响及其可能的机制。方法构建EGFR特异性shRNA表达载体并转染Colo-16细胞,Western blot检测细胞内EGFR,Akt及NF-κB的表达水平;流式细胞仪检测Colo-16细胞的凋亡变化。结果成功构建EGFR特异性shRNA表达载体并转染Colo-16细胞。转染特异性shRNA后,Colo-16细胞EGFR表达量的灰度值为0.42±0.03,与正常对照组(0.94±0.06)相比降低了55.57%,其差异具有统计学意义(P<0.05)。流式分析显示,下调EGFR表达可诱导Colo-16细胞凋亡,其凋亡率为12.65%,与对照组[凋亡率(0.11±0.03)%,(1.86±0.38)%和(2.26±0.25)%]比较差异具有统计学意义(P<0.05)。此外,抑制EGFR表达还下调了PI3K/AKT信号通路蛋白Akt和NF-κB的表达。结论用特异性shRNA抑制EGFR表达可能是一种有潜力的抗皮肤鳞癌的治疗策略。更多还原

【Abstract】 Objective To investigate the effect of specific shRNA on apoptosis by inhibiting the expression of epidermal growth factor receptor( EGFR) in squamous cell carcinoma Colo-16 cells and to discuss its possible mechanism. Methods The EGFR specific shRNA-expressing vector was established,which was then transfected into Colo-16 cells. Western blot was employed for detecting the expression of EGFR,Akt and NF-κB. Flow cytometry was performed to investigate the apoptosis of Colo-16 cells. Results The EGFRspecific shRNA expressing plasmid was constructed and transfected into Colo-16 cells successfully. The amount of EGFR expression was 0. 42 ± 0. 03 in cells transfected with EGFR-specific shRNA expressing plasmid,which decreased approximately 55. 57% compared with 0. 94 ± 0. 06 in normal control cells( P <0. 05). The Colo-16 cells underwent apoptosis( apoptosis ratio = 12. 65%) in the presence of EGFR-specific shRNA,however,the apoptosis ratio of other control groups were( 0. 11 ± 0. 03) %,( 1. 86 ± 0. 38) %and( 2. 26 ± 0. 25) % respectively( P < 0. 05). Furthermore,the expression of Akt and NF-κB proteins,which involve in PI3 K / AKT signal pathway,was also downregulated. Conclusion Inhibition of EGFR expression by using specific shRNA may be a potential therapeutic strategy in the treatment of cutaneous squamous cell carcinoma.更多还原