bcl-XL反义寡核苷酸转染对结肠癌细胞增殖及裸鼠体内人结肠癌移植瘤生长的抑制作用  

Inhibitory effect of bcl-XL antisense oligodeoxynucleotides on the proliferation of colon cancer cells and the growth of human colon cancer xenografts in nude mice

作　　者：张俊东[1] 刘成[1] 白雪峰[2] 

机构地区：[1]黑龙江省大庆油田总医院普外胃肠外科,大庆163001 [2]哈尔滨医科大学附属肿瘤医院结直肠外科,150040

出　　处：《中华实验外科杂志》2017年第8期1322-1325,共4页Chinese Journal of Experimental Surgery

摘　　要：目的 观察bcl-XL反义寡核苷酸转染后,对结肠癌细胞增殖及裸鼠体内人结肠癌移植瘤生长的影响.方法 制备对数生长期的SW620单细胞悬液,裸鼠背侧皮下注射.当移植瘤体积平均50mm时,将裸鼠随机分为3组：对照组、N-ODN组和反义寡核苷酸组,每组5只.反义寡核苷酸组及N-ODN组瘤旁注射2ml/kg体重的反义寡核苷酸或N-ODN,对照组瘤旁注射等量生理盐水,第16天处死裸鼠,取部分肿瘤组织,提取RNA及蛋白,采用阳离子脂质体介导的反义寡核苷酸转染、实时定量聚合酶链反应（Real-time PCR）、Western blot、噻唑蓝（MTT）法、原位缺口末端标记法（TUNEL）等方法观察了bcl-XL反义寡核苷酸转染对结肠癌细胞增殖、凋亡的影响及裸鼠体内人结肠癌移植瘤生长的影响.结果 （1）对细胞增殖活性的抑制率随反义寡核苷酸浓度的增加逐渐增高,具剂量依赖性.当浓度超过200nμmol/L后,对细胞活性的增殖抑制率递增幅度明显减小,差异有统计学意义（t=2.418、1.531和1.838,P=0.003、0.016和0.011）.（2）细胞对照组、空白对照组、N-ODN组及反义寡核苷酸组的mRNA相对表达量分别为0.96±0.25、0.97±0.48、0.78±0.32及0.41±0.13,抑制率分别为0.00%、-6.25%、18.75%及57.29%,与细胞对照组比较,除空白对照组有少量增高外,N-ODN组及反义寡核苷酸组的表达依次减弱,抑制率依次增加.反义寡核苷酸组与细胞对照组、空白对照组及N-ODN组比较,差异有统计学意义（P=0.025）;N-ODN组、空白对照组及细胞对照组间两两比较,差异无统计学意义（P=0.263）.（3）与细胞对照组比较,空白对照组无明显变化;N-ODN组稍有降低;反义寡核苷酸组明显降低（P=0.017）.（4）细胞对照组、空白对照组、N-ODN组及反义寡核苷酸组的阳性细胞的阳性率分别为3.7%、5.0%、7.0%及35.0%.经统计学分析,反义寡核苷酸组与细胞对照组、空白对照组及N-ODN组比较,差异有统�Objective To investigate the effect of bcl-XL antisense oligonucleotide transfection on the proliferation of colon cancer cells and the growth of human colon cancer xenografts in nude mice.Methods A long-term SW620 single cell suspension was prepared for subcutaneous injection in nude mice.Nude mice were randomly divided into 3 groups： control group,N-ODN group and antisense oligonucleotide group （n=5）,with an average of 50 mm3.The antisense oligonucleotide group and N-ODN group were injected with 2 ml/kg antisense oligonucleotide or N-ODN,and the control group was injected with equal amount of normal saline.At 16th day after the operation,the tumor cells were taken out,and some tumor tissues were extracted.RNA and protein were extracted.The cationic liposome mediated transfection of antisense oligonucleotide,real-time quantitative polymerase chain reaction （Real-time PCR）,Western blotting,methyl thiazol tetrazolium （MTT） method and TdT-mediated dUTP nick end labeling （TUNEL） method were applied to observe the effects of bcl-XL antisense oligonucleotide on the proliferation and apoptosis of colorectal cancer cells in vivo and the growth of human colon cancer xenografts.Results （1） The inhibition rate of cell proliferation activity was gradually increased with the increase of antisense oligonucleotide concentration.When the concentration was over 200 nmol/L,the inhibition rate of cell proliferation was significantly decreased with the difference being statistically significant （t=2.418,1.531 and 1.838,P=0.003,0.016 and 0.011）.（2） The relative expression levels in the control group,blank control group,N-ODN group and mRNA antisense oligonucleotides group were 0.96±0.25,0.97±0.48,0.78±0.32 and 0.41±0.13,the inhibition rate was 0.00%,-6.25%,18.75% and 57.29%,respectively.There was significant difference between antisense oligonucleotide group and control group,blank control group or N-ODN group （P=0.025）.（3） Compared with the control group,there was no significant change in t

关 键 词：结肠癌 寡核苷酸类 BCL-XL 增殖 

分 类 号：R735.35[医药卫生—肿瘤]