CAJ | 학술논문

目的探讨微小RNA185(miR-185)对食管癌细胞增殖、迁移的影响及其分子机制.方法采用实时定量PCR检测miR-185在23例(2002至2012年中国医学科学院肿瘤医院胸外科接受手术治疗的经病理诊断证实的食管癌患者)手术切除新鲜冻存的食管鳞癌及其配对的癌旁正常组织中的表达情况；采用xCELLigence RTCA MP系统和Transwell实验分别检测miR-185对食管癌细胞增殖、迁移能力的影响；在食管癌细胞KYSE150中转染miR-185模拟物,采用实时定量PCR检测靶基因Six1的下游基因细胞周期素A1 mRNA水平的变化；在食管癌细胞KYSE30中转染miR-185抑制剂后,通过免疫荧光实验观察靶基因Six1下游基因E-钙黏蛋白表达水平的变化.结果 miR-185在食管鳞癌中的相对表达量较癌旁正常组织低(0.006比0.039,P=0.016)；瞬时转染miR-185模拟物后,食管癌细胞的增殖和迁移能力均下降[KYSE150迁移、KYSE150侵袭的对照组和实验组细胞迁移个数分别为(146±15)和(64±11)个、(110±12)和(67±5)个,均P＜0.05],同时Six1的下游基因细胞周期素A1的mRNA表达相应下调；瞬时转染miR-185抑制剂后,Six1的下游基因E-钙黏蛋白的表达水平明显下降.结论 miR-185能够通过其靶基因Six1的下游基因细胞周期素A1和E-钙黏蛋白抑制食管癌细胞的增殖和迁移.
목적 탐토미소RNA185(miR-185)대식관암세포증식、천이적영향급기분자궤제.방법 채용실시정량PCR검측miR-185재23례(2002지2012년중국의학과학원종류의원흉외과접수수술치료적경병리진단증실적식관암환자)수술절제신선동존적식관린암급기배대적암방정상조직중적표체정황；채용xCELLigence RTCA MP계통화Transwell실험분별검측miR-185대식관암세포증식、천이능력적영향；재식관암세포KYSE150중전염miR-185모의물,채용실시정량PCR검측파기인Six1적하유기인세포주기소A1 mRNA수평적변화；재식관암세포KYSE30중전염miR-185억제제후,통과면역형광실험관찰파기인Six1하유기인E-개점단백표체수평적변화.결과 miR-185재식관린암중적상대표체량교암방정상조직저(0.006비0.039,P=0.016)；순시전염miR-185모의물후,식관암세포적증식화천이능력균하강[KYSE150천이、KYSE150침습적대조조화실험조세포천이개수분별위(146±15)화(64±11)개、(110±12)화(67±5)개,균P＜0.05],동시Six1적하유기인세포주기소A1적mRNA표체상응하조；순시전염miR-185억제제후,Six1적하유기인E-개점단백적표체수평명현하강.결론 miR-185능구통과기파기인Six1적하유기인세포주기소A1화E-개점단백억제식관암세포적증식화천이.
Objective To explore the role and mechanism of microRNA185 (miR-185) on proliferation,migration and invasion of esophageal squamous cell carcinoma (ESCC).Methods Samples were obtained from 23 ESCC patients undergoing surgery whose were confirmed by pathological diagnosis of esophageal carcinoma from 2002 to 2012,at Department of Thoracic Surgery,Cancer Institute and Cancer Hospital,Chinese Academy of Medical Sciences and Peking Union Medical College.Real-time PCR was used to measure the expression of miR-185.The xCELLigence RTCA MP system and Transwell assay were performed to detect the effect of miR-185 on proliferation,migration and invasion of ESCC respectively.After transfectiing of miR-185 mimic into KYSE150,the expression of Sixl's downstream gene cyclin A1 was evaluated by real-time PCR.After transfection of miR-185 inhibitor into KYSE30,the expression of Ecadherin,a downstream protein of Sixl,was observed under confocal microscope.Results The expression level of miR-185 was down-regulated in ESCC compared with adjacent normal tissue (0.006 vs 0.039,P =0.016).After transfection of miR-185 mimic,miR-185 significantly inhibited proliferation,migration and invasion of ESCC.Transwell assay showed,in comparison with the control group,the number of KYSE150 metastatic and invasive cells was respectively decreased(146 ± 15 vs 64 ± 11,110 ± 12 vs 67 ± 5,both P ＜ 0.05).And the expression level of cyclin A1 decreased.After transfection of miR-185 inhibitor,the expression level of E-cadherin decreased.Conclusion miR-185 may inhibit proliferation,migration and invasion of ESCC through its target gene Six1.