论文部分内容阅读
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。
Objective To investigate the effect of epidermal growth factor receptor (EGFR) on the apoptosis of Colo-16 cells by specific short hairpin RNA (shRNA) and its possible mechanism. Methods EGFR-specific shRNA expression vector was constructed and transfected into Colo-16 cells. The expression of EGFR, Akt and NF-κB were detected by Western blot. The apoptosis of Colo-16 cells was detected by flow cytometry. Results EGFR-specific shRNA expression vector was successfully constructed and transfected into Colo-16 cells. After transfected with specific shRNA, the gray value of EGFR expression in Colo-16 cells was 0.42 ± 0.03, which was 55.57% lower than that of the normal control group (0.94 ± 0.06), the difference was statistically significant (P <0.05) . Flow cytometry analysis showed that down-regulation of EGFR expression induced apoptosis of Colo-16 cells with a apoptotic rate of 12.65%, which was significantly higher than that of the control group [apoptosis rate (0.11 ± 0.03)%, (1.86 ± 0.38)% and (2.26 ± 0.25 )%] The difference was statistically significant (P <0.05). In addition, inhibition of EGFR expression downregulated the expression of PI3K / AKT signaling pathway proteins Akt and NF-κB. Conclusion Inhibition of EGFR expression with specific shRNA may be a potential therapeutic strategy for anti-squamous cell carcinoma.