应用shRNA抑制Skp2表达对喉癌化疗的增敏作用研究  

Effect of Skp2 shRNA on the Chemosensitivity of Laryngeal Squamous Carcinoma Cells

作　　者：路秀英[1] 李晓明[1] 

机构地区：[1]解放军白求恩国际和平医院耳鼻咽喉头颈外科-全军耳鼻咽喉头颈外科中心,石家庄050082

出　　处：《临床误诊误治》2014年第9期36-39,共4页Clinical Misdiagnosis & Mistherapy

基　　金：全军医药卫生十一五科技攻关项目(200606M076)

摘　　要：目的探讨shRNA(small hairpin RNA)抑制S期激酶相关蛋白2(S-phase kinase associated protein 2,Skp2)基因表达对化疗后喉癌Hep-2细胞增殖和凋亡的影响。方法将喉癌细胞分为空质粒对照组、Skp2 shRNA组、Skp2 shRNA加化疗组、空质粒加化疗组及化疗组。应用脂质体将Skp2 shRNA转染到人喉癌Hep-2细胞,6 h后加入顺铂,培养48 h后流式细胞术检测Skp2、p27蛋白表达和细胞凋亡,四甲基偶氮唑蓝(MTT)法检测细胞增殖变化,并进行比较。结果 Skp2 shRNA组与空质粒对照组比较,Skp2蛋白表达明显降低,p27蛋白表达明显增高,差异有统计学意义(P<0.01,P<0.05)。Skp2 shRNA加化疗组与空质粒加化疗组比较,各剂量顺铂(0 mg/L、1 mg/L、2 mg/L、3 mg/L)作用下细胞增殖抑制率均明显增高,差异均有统计学意义(P均<0.01);与空质粒加化疗组(2 mg/L顺铂)比较,Skp2 shRNA加化疗组细胞凋亡率明显增加,差异有统计学意义(P<0.01)。结论 Skp2 shRNA可选择性阻断细胞Skp2基因表达,联合化疗可能为喉癌的综合治疗提供新的途径。Objective To investigate the effect of silencing of S-phase kinase associated protein 2 （Skp2） by small hairpin RNA （ shRNA） in combination with chemotherapy on proliferation and apoptosis in laryngeal carcinoma cell line Hep-2 cells. Methods Laryngeal cancer cells were divided into pGPU6 group, Skp2 shRNA group, Skp2 shRNA with chemotherapy group, pGPU6 with chemotherapy group and chemotherapy group. After transfected with Skp2 shRNA, Hep-2 cells were cultured and received cisplatin 6 h after that. Forty-eight hours after cultured, the cell proliferation was assayed by MTT, and apoptosis rate and the expression of Skp2 and p27 protein were analyzed by flow cytometry. Results The expression of Skp2 protein in Skp2 shRNA group was significantly lower than that in pGPU6 group （P〈0. 01）. The expression of p27 protein in Skp2 shRNA group was higher than that in pGPU6 group （P〈0. 05）. The growth inhibition rate in Skp2 shRNA group with cisplatin at 0 mg/L, 1 mg/L, 2 mg/L and 3 mg/L were higher than those in pGPU6 group with cisplatin at 0 mg/L, 1 mg/L, 2 mg/L and 3 mg/L, respectively （P〈0. 01）. The apoptosis rate in Skp2 shRNA with cisplatin group （2 mg/L） was strikingly higher than that in pGPU6 with cisplatin group （2 mg/L）（P〈0. 01）. Conclusion Skp2 shRNA could inhibit the expression of Skp2 gene selectively and combining with chemotherapy might provide a new therapeutic approach to treatment of laryngeal carcinomas.

关 键 词：喉肿瘤 癌 鳞状细胞 S期激酶相关蛋白质类 药物疗法 

分 类 号：R739.65[医药卫生—肿瘤]