miR-124抑制胃癌细胞增殖和侵袭的机制研究  

miR-124 suppresses cell proliferation and invasion in gastric carcinoma and its mechanism

作　　者：谢黎明[1] 贺荣芳[2] 张志伟[3] 唐云云[3] 罗招阳[3] 

机构地区：[1]南华大学附属第一医院肿瘤内科,衡阳421001 [2]南华大学附属第一医院病理科,衡阳421001 [3]南华大学肿瘤研究所

出　　处：《中华肿瘤杂志》2013年第7期497-500,共4页Chinese Journal of Oncology

基　　金：基金项目：国家自然科学基金（31100935）

摘　　要：目的探讨miR-124抑制胃癌细胞增殖和侵袭的机制。方法构建鞘氨醇激酶1（SPHKl）3’UTR-荧光素酶报告载体，通过荧光素酶报告基因实验检测miR-124对SPHKl3’UTR-荧光素酶活性的影响。将miR-124模拟物转染胃癌细胞MGC-803，采用Westernblot检测SPHKl表达水平，采用四甲基偶氮唑蓝（MTT）和Transwell侵袭实验检测miR-124与SPHKlsiRNA对MGC-803细胞生长、侵袭转移能力的影响。结果荧光素报告载体系统证实，SPHKl是miR-124直接调控的靶基因。Westernblot检测结果显示，miR-124可抑制SPHKI蛋白的表达。MTT法检测结果显示，转染12、24、48h后，对照组的A值分别为0．316±0．010、0．429±0．002和0．517±0．007，miR．124模拟物组的A值分别为0．152±0．003、0．216±0．001和0．242±0．020，SPHKlsiRNA组的A值分别为0．167±0．006、0．286±0．011和0．311±0．040，miR-124模拟物组和SPHKIsiRNA组与对照组差异均有统计学意义（均P〈0．05），且具有时间依赖性。Transwell侵袭实验显示，转染24h后，对照组、miR-124模拟物组和SPHKlsiRNA组的穿膜细胞数分别为（100．6±11．3）个、（47．8±6．6）个和（54．6±8．3）个，miR．124模拟物和SPHKlsiRNA能明显减缓MGC-803细胞的侵袭能力，与对照组差异均有统计学意义（均P〈0．05）。结论miR-124可通过靶向调控SPHKl的表达而抑制胃癌细胞的增殖和侵袭能力。Objective To explore the molecular mechanism of miR-124 suppressing the proliferation and invasion of gastric cancer cells. Methods SPHK1 3＇UTR-luciferase vector was constructed and luciferase reporter gene assay was employed to examine the effect of miR-124 on luciferase activity. Human gastric cancer MGC-803 cells were transfected with miR-124 mimics, and then Western blot was performed to detect the expression of SPHK1 protein. Results Luciferase reporter vector system confirmed that SPHK1 was a target gene of miR-124. Western blot showed that the expression of SPHK1 protein was inhibited by miR-124. After transfection of miR-124 mimics or SPHK1 siRNA for 12 h, 24 h and 48 h, respectively, M＇IT assay showed that the A values of the three groups were significantly different （ P 〈 0.05 ）, and it was in a time-dependent manner. After transfection of miR-124 mimics or SPHK1 siRNA for 24 h, transwell invasion assay showed that the number of transmembrane cells was 54.6±8.3 in the SPHK1 siRNA group and 47.8 ±6.6 in the miR-124 mimics group, both were significantly lower than 100.6±11.3 of the control group （ P 〈 0.05 ）, indicating that SPHK1 siRNA can slow down the invasion of MGC-803 cells. Conclusion miR-124 can suppress the cell proliferation and invasion by targeting SPHK1 in gastric carcinoma.

关 键 词：胃肿瘤 miR-124 鞘氨醇激酶1 细胞增殖 肿瘤侵润 

分 类 号：R735.2[医药卫生—肿瘤]