DUOX2对结直肠癌细胞氟尿嘧啶敏感性的影响  

Effect of DUOX2 on sensitivity of colorectal cancer cells to 5-fluorouracil

作　　者：张雪[1] 刘妍[1] 周欣亮[1] 范志松[1] 韩晶[1] 冯莉[1] 左静[1] 王贵英[2,3] ZHANG Xue;LIU Yan;ZHOU Xinliang;FAN Zhisong;HAN Jing;FENG Li;ZUO Jing;WANG Guiying(Department of Medical Oncology,the Fourth Affiliated Hospital of Hebei Medical University,Shijiazhuang 050011,Hebei,China;The Second Department of General Surgery,the Fourth Affiliated Hospital of Hebei Medical University,Shijiazhuang 050011,Hebei,China;Department of Gastrointestinal Surgery,the Third Affiliated Hospital of Hebei Medical University,Shijiazhuang 050051,Hebei,China)

机构地区：[1]河北医科大学第四医院肿瘤内科,河北石家庄050011 [2]河北医科大学第四医院外二科,河北石家庄050011 [3]河北医科大学第三医院胃肠外科,河北石家庄050051

出　　处：《中国肿瘤生物治疗杂志》2020年第12期1365-1371,共7页Chinese Journal of Cancer Biotherapy

基　　金：国家癌症中心肿瘤科研专项课题(No.NCC2017A23)。

摘　　要：目的:探讨双氧化酶2(dual oxidase 2,DUOX2)对结直肠癌(colorectal cancer,CRC)细胞5-氟尿嘧啶(5-fluorouracil,5-FU)药物敏感性的影响。方法:选用CRC细胞系DLD-1、SW480、HCT116、SW620与正常肠上皮细胞株NCM460,用qPCR法检测细胞中DUOX2的表达水平。利用慢病毒感染技术,稳定敲降HT-29与HCT116细胞中DUOX2表达,qPCR法和WB法检测敲降效率。用不同浓度(0、5、10、20、40、80、120μg/ml)5-FU处理sh-Control组与sh-DUOX2组细胞,用CCK-8法、流式细胞术分别检测5-FU对细胞增殖、细胞凋亡和细胞周期的影响。构建裸鼠HT29细胞移植瘤模型,观察DUOX2基因对5-FU疗效的影响。结果:DUOX2 mRNA在CRC细胞中的表达水平显著高于NCM460细胞(P<0.05或P<0.01)。敲降DUOX2基因后,与sh-Control组相比,sh-DUOX2组HT29和HCT116细胞中DUOX2 mRNA和蛋白表达水平明显下降(均P<0.01);细胞对5-FU的敏感性增强,细胞凋亡率明显升高(均P<0.01),G0/G1期比例显著升高、G2期与S期比例显著下降(均P<0.01)。未经5-FU治疗的sh-Control组与sh-DUOX2组裸鼠移植瘤体积及质量比较差异均无统计学意义(均P>0.05),经5-FU治疗的sh-DUOX2+5-FU组移植瘤体积及质量明显低于sh-Control+5-FU组(均P<0.01)。结论:敲降DUOX2基因可显著增强CRC细胞对5-FU的敏感性。Objective:To investigate the effect of double oxidase 2(DUOX2)on the sensitivity of colorectal cancer(CRC)cells to 5-fluorouracil(5-FU).Methods:CRC cell lines DLD-1,SW480,HCT116,SW620 and normal intestinal epithelial cell line NCM460 were selected,and the expression of DUOX2 in these cell lines were detected by qPCR.DUOX2 expression in HT-29 and HCT116 cells was stably knocked down by lentivirus infection technique.The knockdown efficiency was detected by qPCR and WB.Cells in sh-Control and sh-DUOX2 groups were treated with 5-FU at different concentrations(0,5,10,20,40,80,120μg/ml).The effects of 5-FU on cell proliferation,apoptosis and cell cycle were detected by CCK-8 method and flow cytometry.HT29 cell transplanted xenograft model in nude mice was constructed to observe the effect of DUOX2 gene on the treatment efficacy of 5-FU.Results:the expression level of DUOX2 mRNA in CRC cells was significantly higher than that in NCM460 cells(P<0.05 or P<0.01).Compared with sh-Control group,the mRNA and protein expressions of DUOX2 in sh-DUOX2 group were significantly decreased(all P<0.01);the sensitivity of cells to 5-FU was enhanced,the apoptosis rate and the ratio of cells at G0/G1 phase were significantly increased(all P<0.01),and the ratio of cells at G2 and S phase was significantly decreased(all P<0.01).There was no significant difference in tumor volume and mass between sh-Control group and sh-DUOX2 group without 5-FU treatment(all P>0.05),but the volume and mass of transplanted tumor in sh-DUOX2+5-FU group after 5-FU treatment was significantly lower than that in sh-Control+5-FU group(all P<0.01).Conclusion:The sensitivity of CRC cells to 5-FU can be significantly enhanced by knocking down DUOX2 gene.

关 键 词：双氧化酶2 5-氟尿嘧啶 结直肠癌 HT29细胞 HCT116细胞 增殖 凋亡 

分 类 号：R735.3[医药卫生—肿瘤] R730.5[医药卫生—临床医学]