miRNA-199 a和 miRNA-214通过负调控MAPK通路因子抑制宫颈癌细胞生长增殖  

miRNA-199a and miRNA-214 regulate its target genes in MAPK pathway and inhibit cell growth and proliferation of cervical cancer

作　　者：马承宁[1] 王蕊[1] 莫文娟[1] 吕红[1] 

机构地区：[1]复旦大学生命科学学院遗传工程国家重点实验室,上海200433

出　　处：《国际遗传学杂志》2014年第6期269-277,290,共10页International Journal of Genetics

基　　金：国家自然科学基金重大研究计划（91129730）;教育部博士点基金优先发展领域（20110071130007）

摘　　要：目的：研究 miRNA-199a和miRNA-214调控宫颈癌细胞生长增殖的分子机制。方法用细胞生长曲线实验和克隆形成实验研究 miRNA-199a和miRNA-214对宫颈癌细胞系 HeLa 细胞生长增殖的抑制作用；利用信息学手段及双荧光素酶报告基因方法筛选鉴定miRNA-199a和miRNA-214的靶基因，用Real-timePCR和Western印迹方法研究它们对靶基因表达量的影响。结果miRNA-199a和miRNA-214显著抑制了HeLa细胞生长增殖（P ＝0.010、0.036）；鉴定了MAPK通路的KRAS和MKK4（JNKK1／MAP 2K4）是miRNA-199a的靶基因，同时 KRAS 也是 miRNA-214的靶基因；miRNA-199a能够抑制HeLa细胞 KRAS 和 MKK 4的表达（P ＝0.231、0.011），而miRNA-214也能够抑制 KRAS 的表达（P ＝0.029）。结论miRNA-199a和miRNA-214通过调节MAPK信号途径因子的表达，参与了宫颈癌细胞的生长增殖。Objective To investigate the mechanism of regulations of miRNA-199a and miRNA-214 in cell growth and proliferation of cervical cancer .Methods We used cell growth curves and cell clones formation assays to study effects of miRNA-199a and miRNA-214 on growth of HeLa cells; Bioin-formatics tools and dual-luciferase report gene assays were used to screen and identify target genes of miRNA-199a and miRNA-214 in MAPK pathway; Realtime-PCR and Western blotting were used to in-vestigate the inhibitory effects on those target genes of miRNA-199a and miRNA-214 . Results We proved that miRNA-199a , miRNA-214 could inhibit growth and proliferation of HeLa cells （ P =0.0 1 0 , 0.036 ）; we identified miRNA-199a ’ s target genes KRAS and MKK 4 , miRNA-214 ’ s target gene KRAS , and proved the inhibitory effects of miRNA-199a and/or miRNA-214 on expression of those target genes （ P =0.231,0.011 ,0.029） .Conclusion miRNA-199a , miRNA-214 could play im-portant roles in cell growth and proliferation of cervical cancer through affecting MAPK pathway .

关 键 词：宫颈癌 miRNA-199a miRNA-214 MAPK 

分 类 号：R737.33[医药卫生—肿瘤]