c-myc反义寡核苷酸对结肠癌HT-29细胞增殖、凋亡及化疗敏感性的影响  

Effect of c-myc antisense oligodeoxynucleotides on proliferation, apoptosis and chemosensitivity of colorectal cancer cell line HT-29

作　　者：邵国利[1] 吴爱国 焦得闯[1] 纪术峰[1] 

机构地区：[1]南方医科大学附属珠江医院普通外科,广东省广州市510282

出　　处：《世界华人消化杂志》2009年第5期459-463,共5页World Chinese Journal of Digestology

基　　金：广东省科技计划基金资助项目;No.2005B31201011~~

摘　　要：目的:研究体外c-myc反义寡核苷酸(ASODN)对人结肠癌HT-29细胞增殖、凋亡及化疗敏感性的影响.方法:利用脂质体LipofectamineTM2000介导将c-myc ASODN转染入大肠癌HT-29细胞中,逆转录多聚酶链反应(RT-PCR)、Western blot方法检测c-myc基因mRNA及蛋白的表达,MTT、流式细胞仪(FCM)检测c-myc ASODN对人结肠癌HT-29细胞增殖抑制及其对奥沙利铂敏感性影响.结果:转染c-myc ASODN后,HT-29细胞内c-mycm RNA水平显著降低(0.464±0.029vs0.974±0.027,0.945±0.012,均P<0.01).在HT-29细胞中存在分子质量62kDa的特异性条带,与c-myc分子质量相符,ASODN组在PVDF膜上的特异性条带明显弱于对照组;MTT结果显示转染c-myc ASODN48h后的HT-29细胞的增殖速度较对照组细胞明显减慢.FCM显示c-myc ASODN转染后72h后,奥沙利铂+c-myc ASODN组细胞凋亡率显著高于对照组(P<0.05).结论:体外c-myc ASODN可抑制c-myc mRNA及蛋白的表达,阻断c-myc可抑制HT-29细胞增殖并增强其对奥沙利铂的敏感性,可能为大肠癌的基因治疗提供新的靶点.AIM： To investigate the effects of c-myc antisense oligodeoxynucleotides （ASODN） on the proliferation, apoptosis and chemosensitivity to human colorectal cancer cell line HT-29 in vitro. METHODS： c-myc ASODN and scrambled oligo- deoxynueleotide （SODN） were transfected with lipofectamineTM 2000 into human colorectal cancer cell line HT-29. The expression of c-myc mRNA and protein in human colorectal cancer cell line HT-29 was detected by semi quantitive reverse transcription-polymerase chain reaction （RT-PCR） and Western blot, respectively. The effects of the c-myc ASODN on the proliferation and sensitivity to oxaliplatin of HT-29 cell line were studied by MTT assay and flow cytometry （FCM）. RESULTS： The expression of c-myc mRNA level in HT-29 was down-regulated remarkably between the antisense group and control group （0.464 ± 0.029 vs 0.974 ±0.027, 0.945 ±0.012, both P 〈 0.01）. Protein expression of c-myc in HT-29 cells was confirmed and it was clear that specific band on PVDF membrane in ASODN group was significantly weakened than those in control groups; MTT and FCM showed the proliferation of HT-29 transfected with c-myc ASODN was retarded obviously in contrast to control groups after 48 hours＇ transfection （P 〈 0.05）. Compared with the control groups after 72 hours＇ transfection, the percentage of the survival cells signifi- cantly decreased in the presence of both oxaliplatin and c-myc ASODNs （P 〈 0.05）. CONCLUSION： The c-myc ASODN can inhibit c-myc expression and inhibit the proliferation of colonic cancer HT-29 cells. Combination of c-myc ASODN and oxaliplatin might improve the treatment outcome of colorectal carcinoma. c-myc may be a potential target of gene therapy for human colorectal cancer.

关 键 词：C-MYC 反义寡核苷酸 反义治疗 HT-29 细胞 大肠癌 

分 类 号：R735.35[医药卫生—肿瘤]