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目的探讨microRNA-133b(miR-133b)对人结直肠癌裸鼠皮下种植瘤生长的抑制作用,体内实验进一步验证miR-133b是一种新的抑癌基因。方法 30只裸鼠按随机数字表法分为接种人结直肠癌细胞SW-620细胞组、接种SW-620-HK细胞组及接种SW-620-133b细胞组,每组10只,建立裸鼠皮下种植瘤模型,记录并比较各组种植瘤生长情况,TUNEL法检测种植瘤细胞凋亡,荧光实时定量PCR检测各组种植瘤中miR-133b表达,Westernblot检测c-met表达。结果各组裸鼠皮下接种肿瘤细胞5d后均成功致瘤,成瘤率100%;SW-620-133b组种植瘤生长速度显著低于SW-620与SW-620-HK组(P<0.05);终点时间第30天时,SW-620组裸鼠皮下种植瘤的体积为(3.56±0.79)cm3,SW-620-HK组为(3.24±0.68)cm3,显著大于SW-620-133b组肿瘤体积(0.85±0.14)cm3(P<0.05);SW-620-133b组种植瘤质量为(1.15±0.16)g,明显低于SW-620组(4.02±0.64)g及SW-620-HK组(3.81±0.61)g(P<0.05);SW-620-133b组肿瘤细胞凋亡率(30.28±3.47)%显著高于对照组SW-620-HK(7.26±0.84)%及空白组SW-620(7.96±0.92)%(P<0.05);SW-620-133b组种植瘤中c-met表达量(0.19±0.02)较SW-620组(0.52±0.04)及SW-620-HK组(0.51±0.04)显著下降(P<0.05)。结论外源性高表达miR-133b可显著抑制人结直肠癌裸鼠皮下种植瘤的生长。
Objective To investigate the inhibitory effect of microRNA-133b (miR-133b) on the growth of human colorectal cancer xenografts in nude mice, and to further verify that miR-133b is a novel tumor suppressor gene in vivo. Methods Thirty nude mice were randomly divided into three groups: SW-620 cells inoculated with human colorectal cancer cells, SW-620-HK cells inoculated with SW-620-HK cells, and SW-620-133b cells inoculated with SW-620-133b cells The growth of implanted tumor was recorded and compared. TUNEL method was used to detect the apoptosis of implanted tumor cells. Real-time quantitative PCR was used to detect the expression of miR-133b in each group. Western blot was used to detect the expression of c-met. Results The nude mice were successfully inoculated with tumor cells 5 days after the inoculation of nude mice, and the tumorigenic rate was 100%. The growth rate of SW-620-133b group was significantly lower than that of SW-620 and SW-620-HK groups (P <0.05) (3.56 ± 0.79) cm3 in SW-620 group and (3.24 ± 0.68) cm3 in SW-620-HK group were significantly higher than those in SW-620-133b group on the 30th day (0.85 ± 0.14) cm3 (P <0.05). The tumor mass in SW-620-133b group was (1.15 ± 0.16) g, which was significantly lower than that in SW-620 group (4.02 ± 0.64) g and SW-620-HK group 3.81 ± 0.61) g (P <0.05). The apoptosis rate of SW-620-133b group was significantly higher than that of SW-620-HK group (7.26 ± 0.84)% and SW-620 (7.96 ± 0.92)% (P <0.05). The expression of c-met in SW-620-133b group was significantly higher than that in SW-620 group (0.52 ± 0.04) and SW-620-HK group ± 0.04) (P <0.05). Conclusion Exogenous high expression of miR-133b can significantly inhibit the growth of human colorectal cancer subcutaneous tumor in nude mice.