miR-200c在CIK细胞诱导胃癌细胞凋亡中的作用  

The role of miR-200c in the apoptosis of gastric cancer cell induced by CIK cells

作　　者：杜宪红[1] 邢召全[2] 焦晋山[1] 

机构地区：[1]山西医科大学第一医院输血科,太原030001 [2]山东大学齐鲁医院泌尿外科,济南250012

出　　处：《山东大学学报（医学版）》2013年第6期49-52,56,共5页Journal of Shandong University:Health Sciences

基　　金：中国博士后科研基金(2012M521346);山东省博士后创新基金(201203041)

摘　　要：目的观察细胞因子诱导的杀伤细胞(CIK)对胃癌细胞凋亡的影响,探讨微RNA-200c(miR-200c)在此过程中的作用。方法实时荧光定量RT-PCR检测胃癌细胞(MKN-45、BGC-803)miR-200c表达,流式细胞术检测胃癌细胞凋亡。构建包含miR-200c作用位点的psiCHECK2-FAP-1 3’-UTR,转染胃癌细胞,双荧光素酶报告基因试剂盒检测胃癌细胞Fas相关磷酸酯酶-1(FAP-1)荧光素酶活性。结果 CIK细胞促进MKN-45细胞miR-200c表达[(2.10±0.25)倍,P<0.05]及凋亡(P<0.01),而miR-200c inhibitor能够拮抗此过程(P<0.05)。另外,miR-200c mimics诱导胃癌细胞凋亡(P<0.05),BGC-803细胞也发现相似趋势。miR-200c直接作用于FAP-1基因3’-UTR区,且CIK细胞降低胃癌细胞荧光素酶活性[MKN-45细胞:(49.67±2.36)%,P<0.01;BGC-803细胞:(43.86±4.41)%,P<0.01]。结论 CIK细胞通过上调miR-200c促进胃癌细胞凋亡,为CIK细胞治疗胃癌提供新的数据。Objective To observe the effect of cytokine-induced killer cells （CIK cells） on the apoptosis of gastric cancer （GC） cells, and explore the role of microRNA-200c （miR-200c） in this process. Methods MiR-200c was determined by real time quantitative PCR, and the apoptosis was detected by flow cytometry in GC cells （MKN-45 and BGC803）. psiCHECK2-FAP-1 3’-UTR containing the binding site of miR-200c was constructed, and then transfected into GC cells. Luciferase activity of FAP-1 （Fas-associated phosphatase-1） was determined with the assistance of dual luciferase report system. Results CIK cells promoted GC cell apoptosis （P〈0.01） and the expression of miR-200c [（2.10±0.25） fold, P〈0.05], and miR-200c inhibitor might block this process （P〈0.05）. Moreover, miR-200c mimics induced GC cell apoptosis （P〈0.05）. The similar trend was also observed in BGC-803 cells. MiR-200c targeted at the site of FAP-1 3’-UTR, and CIK cells decreased the luciferase activity of GC cells [MKN45： （49.67±2.36）%, P〈0.01; BGC803： （43.86±4.41）%, P〈0.01]. Conclusion CIK cells could enhance GC cell apoptosis by upregulating miR-200c, which may provide a new database to elucidate GC therapy using CIK cells.

关 键 词：胃肿瘤 CIK细胞 细胞凋亡 微RNA-200c FAS相关磷酸酯酶-1 

分 类 号：R735.2[医药卫生—肿瘤]