小分子RNA干扰靶向巨噬细胞移动抑制因子对大肠癌影响的实验研究  

The effect of siRNA targeting MIF on the growth of colorectal cancer xenografts and the life quality of tumor-bearing mice

作　　者：王亚敏[1] 王丽京[1] 杨荣娇[1] 蔡洁毅[1] 吴礼浩[1] 何兴祥[1] 

机构地区：[1]广东药学院附属第一医院消化内科,广州510080

出　　处：《中华普通外科杂志》2011年第5期376-380,共5页Chinese Journal of General Surgery

基　　金：国家自然科学基金（30470435）;广东省自然科学基金（9251065005000001）;广东省科学技术厅科技计划项目基金（2008A030201001）;广东省高等学校高层次人才项目（粤教师函[2010]79号）

摘　　要：目的分析靶向巨噬细胞移动抑制因子的小分子干扰RNA（MIFsiRNA）对大肠癌生长与荷瘤小鼠生存质量的影响，并探讨其作用机制。方法利用盲肠造疝原位接种瘤块法建立大肠癌动物模型。将成功建模后的BALB／C小鼠随机分为3组，每组10只，每周2次分别给予DEPC水、MIFsiRNA（0．15nmol／g）和非特异性siRNA（0．15nmol／g）瘤内注射，每天称量各组小鼠的饮水、饮食量和体重，每周测量1次肿瘤体积。4周后处死小鼠，称取肿瘤重量，用ELISA法和免疫组织化学法检测小鼠血清和组织中的MIF水平，分光光度法检测肿瘤组织中Caspase-3蛋白的表达，TUNEL法检测肿瘤组织中凋亡细胞数。结果MIFsiRNA干预组小鼠血清中MIF表达比其他两组低[（22±6）ng／m1）比（32±8）ng／ml、（33±8）ng／ml，P〈0．01]，组织中MIF阳性细胞数比其他两组少[（85±20）／500个比（423±23）／500个、（442±1）／500个，P〈0．01]；干预后第3周与第4周MIFsiRNA干预组小鼠肿瘤体积均明显小于其他两组（P〈0．01）。处死小鼠后肿瘤重量也明显轻于其他两组[（1．93±0．21）g比（4．40±0．30）g、（5．25±0．44）g，P〈0．01]；建模后15～31d饮水量较其他两组多（P〈0．01），建模后8～31d饮食量也较其他两组多（P〈0．01），3组小鼠体重变化之间相比差异无统计学意义（P〉0．05）；MIFsiRNA干预组小鼠肿瘤组织中Caspase-3蛋白表达比其他两组高[（0．74±0．06）μg比（0．57±0．08）μg、（0．56±0．02）μg，P〈0．01]，凋亡细胞数较其他两组多[（12±2）／100个比0、0，P〈0．01]。结论敲低MIF基因表达可以抑制大肠癌的生长，提高荷瘤小鼠的生存质量，其可能的作用机制是激活Caspase-3促进细胞凋亡。Objeetive Toanalyze the effect of siRNA targeting MIF（MIFsiRNA） on the growth of colorectal cancer xenografts and the life quality of tumor-bearing mice. Methods BALB/C mouse model carting colorectal cancer was established. Thirty mice were divided into three groups randomly and managed respectively with intratumor injection of DEPC water, MIFsiRNA（0. 15 nmol/g） and non-specific siRNA （0. 15 nmol/g） ,respectively twice a week for consecutively 4 weeks. Drinking water, fodder consumed and body weight was recorded daily, and tumor volume was measured once a week. Mice were sacrificed after four weeks. ELISA and immunohistochemistry were used to detect the expression of MIF in serum and in tumor tissues. Spectrophotometric detection was used to detect caspase-3 protein. TUNEL was used to detect apoptotic cells. Results MIF expression in serum in MIFsiRNA group was lower than the other two groups [ （ 22 ± 6） ng/ml vs （32 ± 8 ） ng/ml and （ 33 ± 8 ） ng/ml, P 〈 0. 01 ） ; MIF expression in tissues was less than the other two groups [ （ 85±20 ）/500 vs. （ 423± 23 ）/500 and （ 442 ±31 ）/500, P 〈 0. 01 ] ; Tumor was smaller than the other two groups at third and fourth week （P 〈 0. 01 ） ; Tumor weight was significantly less than the other two groups [ （ 1.93 ±0. 21 ） g vs （4. 40±0. 30） g and （5.25 ± 0.44） g, P 〈 0. 01 ] ; Mice in MIFsiRNA group were healthier than the other two groups as judged by water and fodder consumption （P 〈 0. 01 ）, while weight change was not significantly different among the three groups （ P 〉 0.05 ）. Caspase-3 protein in tissues was higher than the other two groups [ （ 0. 74 ± 0. 06 ） μg vs （ 0. 57 ± 0. 08 ） txg and （0. 56±0. 02） μg,P 〈0. 01 ] ;Apoptosis cells in tissues were higher than the other two groups [ （ 12 ± 2） / 100个 vs 0 and 0,P 〈0. 01 ]. Condttsions Knockdowning MIF gene expression inhibits the growth of colorectal cancer xenografts and improves life quality of tumor-bearing mic

关 键 词：结肠肿瘤 RNA 小分子干扰 巨噬细胞游走抑制因子 细胞凋亡 疾病 模型 动物 

分 类 号：R735.34[医药卫生—肿瘤]