5-FU联合pE9-Endostatin对喉癌的抑瘤效应

Antitumor effects of 5-FU combined with pE9-Endostatin on aryngeal carcinoma

目的研究5-FU联合pE9-Endostatin(pE9-En)对喉癌的抑瘤效应。方法体外培养人喉癌Hep-2细胞,通过脂质体将pE9-En转染至细胞株中,采用MTT法测定细胞活力,并计算其半数抑制浓度IC_(50);pE6-EGFP、pE9-EGFP、pE12-EGFP转染至细胞株,流式检测5-FU诱导喉癌Hep-2细胞EGFP表达。将细胞分成pE9组、pE9+5-FU组、pE9-En组及pE9-En+5-FU组,分别采用MTT法、流式细胞术和细胞克隆技术检测细胞增殖和凋亡;将人喉癌Hep-2细胞接种于裸鼠右下肢,建立喉癌移植瘤模型,将16只移植瘤接种成功的裸鼠随机平均分成pE9组、pE9+5-FU组、pE9-En组及pE9-En+5-FU组,检测各组肿瘤体积及质量。结果pE9-En+5-FU组与pE9组、pE9+5-FU组、pE9-En组相比,喉癌细胞增殖能力明显减弱(P<0.05),克隆形成率降低(P<0.05),早期凋亡率明显增加(P<0.05),裸鼠瘤重及肿瘤体积减小(P<0.05)。结论5-FU联合pE9-En抑制喉癌细胞增殖,促进其凋亡,并能抑制小鼠喉癌移植瘤生长。

Objective To investigate the effects of the combined treatment with pE9-Endostatin and 5-FU on laryngeal carcinoma HEP-2 in vitro and in vivo.Methods Human laryngeal carcinoma Hep-2 cells were cultured in vitro,and pE9-En was transfected into the cell line by liposome.The cell viability was de⁃termined by MTT assay,and the 50%inhibitory concentration IC50 was calculated.pE6-EGFP,pE9-EG⁃FP and pE12-EGFP were transfected into the cell lines,and the expression of EGFP in Hep-2 cells in⁃duced by 5-FU was detected by flow cytometry.The cells were divided into pE9 group,pE9+5-FU group,PE9-En group and PE9-En+5-FU group,cell proliferation and apoptosis were detected by MTT assay,flow cytometry and cell cloning technique,respectively.The transplantation tumor models were established by inoculating the human HEP-2 cells into the right lower limb of nude mice.Sixteen nude mice that were successfully inoculated with transplanted tumor were divided equally into pE9 group,pE9+5-FU group,PE9-En group and PE9-En+5-FU group.The antitumor effect of tumor volume and weight on laryngeal carcinoma was detected.Results Compared with pE9,pE9+5-FU and PE9-EN groups,the proliferation a⁃bility of laryngeal carcinoma cells in PE9-EN+5-FU group was significantly decreased(P<0.05),the clone formation rate was significantly decreased(P<0.05),the early apoptosis rate was significantly in⁃creased(P<0.05),and the tumor weight and tumor volume of nude mice were decreased(P<0.05).Conclusion 5-FU combined with pE9-En can inhibit laryngeal carcinoma cell proliferation and promote apopto⁃sis and inhibit the growth of mouse laryngeal carcinoma xenograft.