表皮生长因子受体特异性shRNA对Colo-16细胞凋亡的影响及其机制  

The Effect of Epidermal Growth Factor Receptor-Specific shRNA on Apoptosis and Its Mechanism in Colo-16 Cells

作　　者：赵东利 魏志平 李竹茜 张豫凤 王慧 

机构地区：[1]山东省青岛市城阳区人民医院皮肤科,山东青岛266109 [2]Department of Dermatology,Affiliated Hospital of Xuzhou Medical College

出　　处：《中国皮肤性病学杂志》2016年第6期559-563,共5页The Chinese Journal of Dermatovenereology

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。Objective To investigate the effect of specific shRNA on apoptosis by inhibiting the expression of epider- mal 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-KB. Flow cytometry was performed to investigate the apoptosis of Colo-16 cells. Results The EGFR- specific shRNA expressing plasmid was constructed and transfected into Colo-16 cells successfully. The a- mount of EGFR expression was O. 42 ＋ 0. 03 in cells transfected with EGFR-specific shRNA expressing plas- mid,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-spe- cific 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-KB proteins, which involve in PI3K/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.

关 键 词：表皮生长因子受体(EGFR) SHRNA 皮肤鳞状细胞癌 Colo一16细胞 凋亡 

分 类 号：R751[医药卫生—皮肤病学与性病学]