低频超声联合5-FU对结肠癌HT-29细胞生物学特性的影响  

Effect of low frequency ultrasound combined with 5-FU on biological characteristics of colon cancer HT-29 cells

作　　者：李艳 罗栩伟 朱冬梅 杨姣 张英娟 张慧 陈思佳 刘刚[1] 刘学彬 LI Yan;LUO Xuwei;ZHU Dongmei;YANG Jiao;ZHANG Yingjuan;ZHANG Hui;CHEN Sijia;LIU Gang;LIU Xuebin(Department of Ultrasound,Second Clinical College of North Sichuan Medical College,Nanchong Central Hospital,Nanchong 637000,China;Department of Orthopaedics,Second Clinical Medical College of North Sichuan Medical College,Nanchong Central Hospital)

机构地区：[1]南充市中心医院,川北医学院第二临床医学院超声科,南充637000 [2]南充市中心医院,川北医学院第二临床医学院骨科

出　　处：《山西医科大学学报》2021年第1期20-26,共7页Journal of Shanxi Medical University

基　　金：四川省卫生和计划生育委员会科研课题资助项目(18PJ149);四川省教育厅科研计划项目(17ZB0176);川北医学院科研发展计划资助项目(CBY17-A-YB19,CBY17-A-YB50);南充市市校合作科研专项基金资助项目(19SXHZ0119,19SXHZ0237)。

摘　　要：目的探索低频超声联合5-FU对结肠癌HT-29细胞及其干细胞增殖、凋亡的影响。方法培养人结肠癌细胞系HT-29及其肿瘤干细胞球,随机分为4组并给予相应处理:对照组、低频超声组(30 kHz,37℃,超声1 min)、5-氟尿嘧啶(5-FU)组(药物浓度为5μg/ml)和联合组(低频超声+5-FU)。通过CCK-8实验检测不同处理对细胞活性的影响;流式细胞仪检测细胞凋亡率;qRT-PCR和Western blot检测凋亡相关基因的表达情况。结果与对照组和单独处理组相比,低频超声和5-FU联合处理组细胞增殖明显受到抑制,肿瘤干细胞球的数量、活性也明显降低,干性标志物CD133、CD44和Sox2表达下调,并且凋亡率增加,凋亡相关蛋白PARP裂解片段(cleaved-PARP)表达上调,抑凋亡蛋白Bcl-2表达下调(均P<0.05)。结论低频超声可增强5-氟尿嘧啶对结肠癌细胞活性抑制作用,并且低频超声联合5-氟尿嘧啶可以通过降低结肠癌干细胞的干性而促进诱导结肠癌干细胞凋亡。Objective To explore the effect of low frequency ultrasound combined with 5-FU on colon cancer HT-29 cells and their stem cells.Methods Human colorectal cancer cell line HT-29 and its tumor stem cell spheres were randomly divided into four groups:control group,low frequency ultrasound group(30 kHz,37℃,ultrasound for 1 min),5-fluorouracil group(5μg/ml 5-FU)and combination group(LFUS+5-FU).CCK-8 assay was used to detect the cell viability.Flow cytometry was used to detect the rate of apoptosis.QRT-PCR and Western blot were used to detect the expression of apoptosis-related genes.Results Compared with control group and single treatment group,the cell proliferation was significantly inhibited,and the number and the activity of tumor stem cell spheres were significantly decreased,the expression of dry markers was down-regulated,and the apoptosis rate was increased and the expression of apoptosis-related protein PARP was up-regulated in combination group(all P<0.05).Conclusion Low-frequency ultrasound can enhance the inhibitory effect of 5-fluorouracil on the activity of colon cancer cells,and low-frequency ultrasound combined with 5-fluorouracil can promote the induction of apoptosis of colon cancer stem cells by reducing the dryness of colorectal cancer stem cells.

关 键 词：低频超声 结肠癌 肿瘤干细胞 化疗 5-氟尿嘧啶 

分 类 号：R735.3[医药卫生—肿瘤]