兰州大学学报（医学版）
蘭州大學學報（醫學版）
란주대학학보（의학판）
JOURNAL OF LANZHOU UNIVERSITY(MEDICAL SCIENCES)
2015年
3期
8-13
,共6页
叶小娟%蔡慧%何斌%高勇
葉小娟%蔡慧%何斌%高勇
협소연%채혜%하빈%고용
肝癌%微小RNA-320a%FoxM1%细胞迁移
肝癌%微小RNA-320a%FoxM1%細胞遷移
간암%미소RNA-320a%FoxM1%세포천이
hepatocellular carcinoma%microRNA-320a%FoxM1%cell migration
目的：探讨微小RNA-320a (miR-320a)在肝癌细胞中对癌基因FoxM1的靶向调控作用以及对肝癌细胞迁移能力的影响，为肝癌治疗提供新的靶点。方法通过信息学工具网站http：//www.microRNA.org找到可能靶向调控FoxM1表达的微小RNA；利用蛋白印记和双荧光素酶报告基因试验验证miR-320a对FoxM1的调控作用；采用Transwell小室分析miR-320a和FoxM1对肝癌细胞迁移的影响。结果信息学分析表明，miR-320a在FoxM1的3'端非翻译区存在2个潜在结合位点。蛋白印迹实验显示miR-320a降低FoxM1在肝癌细胞中的蛋白水平，双荧光素酶报告基因试验显示miR-320a可以调控FoxM1表达。miR-320a类似物对FoxM1表达有下调作用（P<0.01），miR-320a抑制物对FoxM1表达有上调作用（P<0.05）。Transwell试验表明miR-320a类似物可抑制肝癌细胞株Huh7、HCC-LM3的迁移能力，miR-320a抑制物可增加Huh7、HCC-LM3的迁移能力。当使用小干扰RNA下调FoxM1在这些细胞株中的表达后，miR-320a类似物和miR-320a抑制物对肝癌细胞迁移能力的调控作用明显减弱。结论 miR-320a有抑制肝癌细胞迁移的作用，这种抑制作用是通过靶向调控癌基因FoxM1来实现的。
目的：探討微小RNA-320a (miR-320a)在肝癌細胞中對癌基因FoxM1的靶嚮調控作用以及對肝癌細胞遷移能力的影響，為肝癌治療提供新的靶點。方法通過信息學工具網站http：//www.microRNA.org找到可能靶嚮調控FoxM1錶達的微小RNA；利用蛋白印記和雙熒光素酶報告基因試驗驗證miR-320a對FoxM1的調控作用；採用Transwell小室分析miR-320a和FoxM1對肝癌細胞遷移的影響。結果信息學分析錶明，miR-320a在FoxM1的3'耑非翻譯區存在2箇潛在結閤位點。蛋白印跡實驗顯示miR-320a降低FoxM1在肝癌細胞中的蛋白水平，雙熒光素酶報告基因試驗顯示miR-320a可以調控FoxM1錶達。miR-320a類似物對FoxM1錶達有下調作用（P<0.01），miR-320a抑製物對FoxM1錶達有上調作用（P<0.05）。Transwell試驗錶明miR-320a類似物可抑製肝癌細胞株Huh7、HCC-LM3的遷移能力，miR-320a抑製物可增加Huh7、HCC-LM3的遷移能力。噹使用小榦擾RNA下調FoxM1在這些細胞株中的錶達後，miR-320a類似物和miR-320a抑製物對肝癌細胞遷移能力的調控作用明顯減弱。結論 miR-320a有抑製肝癌細胞遷移的作用，這種抑製作用是通過靶嚮調控癌基因FoxM1來實現的。
목적：탐토미소RNA-320a (miR-320a)재간암세포중대암기인FoxM1적파향조공작용이급대간암세포천이능력적영향，위간암치료제공신적파점。방법통과신식학공구망참http：//www.microRNA.org조도가능파향조공FoxM1표체적미소RNA；이용단백인기화쌍형광소매보고기인시험험증miR-320a대FoxM1적조공작용；채용Transwell소실분석miR-320a화FoxM1대간암세포천이적영향。결과신식학분석표명，miR-320a재FoxM1적3'단비번역구존재2개잠재결합위점。단백인적실험현시miR-320a강저FoxM1재간암세포중적단백수평，쌍형광소매보고기인시험현시miR-320a가이조공FoxM1표체。miR-320a유사물대FoxM1표체유하조작용（P<0.01），miR-320a억제물대FoxM1표체유상조작용（P<0.05）。Transwell시험표명miR-320a유사물가억제간암세포주Huh7、HCC-LM3적천이능력，miR-320a억제물가증가Huh7、HCC-LM3적천이능력。당사용소간우RNA하조FoxM1재저사세포주중적표체후，miR-320a유사물화miR-320a억제물대간암세포천이능력적조공작용명현감약。결론 miR-320a유억제간암세포천이적작용，저충억제작용시통과파향조공암기인FoxM1래실현적。
Objective To investigate whether MicroRNA-320a (miR-320a) play a role in regulating the migra-tion of hepatocellular carcinoma cell by targeting FoxM1and provide a new target for liver cancer treatment. Methods Bioinformatics tool, microRNA.org was used to find the microRNA which can targeted FoxM1. Western blot and dual luciferase reporter experiments was employed to test the regulation of miR-320a on FoxM1. Transwell chamber was utilized to analyze the effects of miR-320a on the migration of hepatocellular carcinoma cells and analyze the functional relevance between miR-320a and FoxM1 in this process. Results Bioinformatics analysis showed that miR-320a has two potential binding sites in the 3'untranslated region of FoxM1, and the Western blot results showed miR-320a reduced FoxM1 protein levels in liver cancer cells, and double-luciferase reporter experiments further demonstrated that miR-320a could directly regulate FoxM1 expression in post-transcriptional manner. miR-320a mimics can downregulate the expression of FoxM1 (P<0.01), and miR-320a inhibitor can upregulate the expression of FoxM1 (P<0.05). Transwell experiments indicated that miR-320a mimic significantly inhibited the migration of hepatocellular carcinoma cell line-Huh7 and HCC-LM3, on the contrary, miR-320a inhibitor can significantly promoted the migration of hepato-cellular carcinoma cell line. In addition, the functions of miR-320a mimic and inhibitor on liver cancer cells were significantly weakened, when FoxM1 was downregulated by small interfering RNA in these cell lines. Conclusion miR-320a suppresses the migration of liver cancer cells by directly targeting oncogene FoxM1.
