miR-143靶向抑制己糖激酶-2治疗乳腺癌的实验研究  

Experimental study on targeting and inhibiting hexokinase 2 by miR-143 for treatment of breast cancer

作　　者：苗莹[1] 张凌飞[2] 梁胜[3] 石朔[1] 刘默芳[2] 李彪[1] 

机构地区：[1]上海交通大学医学院附属瑞金医院核医学科,上海200025 [2]中国科学院上海生命科学研究院生物化学与细胞生物学研究所,上海200031 [3]上海交通大学医学院附属新华医院核医学科,上海200092

出　　处：《上海交通大学学报（医学版）》2014年第8期1107-1112,共6页Journal of Shanghai Jiao tong University:Medical Science

基　　金：国家自然科学基金(81271610)~~

摘　　要：目的在乳腺癌细胞MDA-MB-231中验证miR-143靶向抑制己糖激酶-2(HK2)改变其能量代谢达到治疗作用,并探讨18F-氟代脱氧葡萄糖(18F-FDG)和3'-脱氧-3'-18F-氟代胸苷(18F-FLT)在评估其疗效中的价值。方法实验分为三组,实验组转染miR-143模拟物(miR-143 mimic),阴性对照组转染无义序列小RNA,空白组不做任何处理。通过qRT-PCR和Western blot分别检测治疗前后HK2 mRNA和蛋白表达水平的改变,检测葡萄糖代谢和乳酸生成速率的变化,MTT法检测细胞增殖活性的变化,测定细胞对18F-FDG和18F-FLT摄取的动态变化,判断两种示踪剂在评估其疗效中的价值。结果实验组HK2 mRNA和蛋白的表达均降低;葡萄糖消耗和乳酸生成速率、细胞增殖活性较阴性对照组明显下降。18F-FDG与18F-FLT分别孵育细胞30、60、90和120 min后,摄取呈时间依赖性递增的趋势,并且各时间点的18F-FLT摄取率均低于18F-FDG。30 min时,实验组18F-FDG摄取率明显低于阴性对照组(21.81±2.75 vs.36.71±4.36),差异具有统计学意义(P<0.05),此时两组摄取18F-FLT无明显差异(12.03±1.53 vs.15.23±2.31,P>0.05)。60 min时,18F-FDG与18F-FLT在两组中的摄取率均出现统计学差异(P<0.05)。结论 miR-143能靶向抑制HK2改变其能量代谢达到治疗作用,18F-FDG在评估其疗效中具有更好的价值。Objective To verify the therapeutic effect of miR-143 on breast cancer cell line MDA-MB-231 by targeting and inhibiting hexokinase 2 （HK2） and changing its energy metabolism and to explore the value of 18F- FDG and 18F-FLT for evaluating the therapeutic effectiveness. Methods Breast cancer cells were divided into three groups, i.e. the test group （ transfected by the miR-143 mimic）, the negative control group （ transfected by scramble sequence of small RNA）, and the blank group （no treatment）. Variations of expressions of mRNA and protein level of HK2 before and after the treatment were detected by qRT-PCR and Western blot. The variations of rates of glucose metabolism and lactic acid production were detected. The variations of proliferative ability of tumor cells were detected by the MTT. The dynamic variations of laF-FDG and 18F-FLT uptake were measured to determine the value of two tracers for evaluating the therapeutic effectiveness. Results The expressions of mRNA and protein level of HK2 of the test group decreased. Compared to the negative control group, the rates of glucose metabolism and lactic acid production and the proliferative ability significandydecreased in the test group. After cells were cultured by 18F-FDG and ISF-FLT for 30, 60, 90, and 120 min, the uptake of 18F-FDG and 18F-FLT showed time-dependent increase and the uptake of 18F-FDG was lower than that of 18F-FLT at each time point. At 30 min, the uptake of 18F-FDG of the test group was significandy lower than that of the negative control group （21.81 ± 2.75 vs 36.71± 4.36）. The difference was statistically significant （P〈0.05）. The uptake of aSF-FLT of two groups showed no significant difference （12.03 ±1.53 vs 15.23±2.31, P〉0.05）. At 60 rain, the differences of the uptake of 18F-FDG and ISF-FLT of two groups were statistically significant （P〈0.05）. Conclusion The therapeutic effect of miR-143 is achieved by targeting and inhibiting HK2 and changing its energy metabolism, 18F-FDG is better for evalu

关 键 词：靶向治疗 乳腺癌 microRNA 18F-FDG 18F-FLT 

分 类 号：R737.9[医药卫生—肿瘤]