国际遗传学杂志
國際遺傳學雜誌
국제유전학잡지
INTERNATIONAL JOURNAL OF GENETICS
2014年
6期
269-277,290
,共10页
马承宁%王蕊%莫文娟%吕红
馬承寧%王蕊%莫文娟%呂紅
마승저%왕예%막문연%려홍
宫颈癌%miRNA-1 9 9 a%miRNA-2 1 4%MAPK
宮頸癌%miRNA-1 9 9 a%miRNA-2 1 4%MAPK
궁경암%miRNA-1 9 9 a%miRNA-2 1 4%MAPK
Cervical cancer%miRNA-1 9 9 a%miRNA-2 1 4%MAPK
目的:研究 miRNA-199 a 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-199 a 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-199 a 和 miRNA-214的靶基因,用 Real-time PCR 和 Western 印迹方法研究它们对靶基因表达量的影响。结果 miRNA-199 a 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 MKK 4( JNKK 1/MAP 2 K 4)是 miRNA-199 a 的靶基因,同时 KRAS 也是 miRNA-214的靶基因;miRNA-199 a 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-199 a 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。
目的:研究 miRNA-199 a 和 miRNA-214調控宮頸癌細胞生長增殖的分子機製。方法用細胞生長麯線實驗和剋隆形成實驗研究 miRNA-199 a 和 miRNA-214對宮頸癌細胞繫 HeLa 細胞生長增殖的抑製作用;利用信息學手段及雙熒光素酶報告基因方法篩選鑒定 miRNA-199 a 和 miRNA-214的靶基因,用 Real-time PCR 和 Western 印跡方法研究它們對靶基因錶達量的影響。結果 miRNA-199 a 和 miRNA-214顯著抑製瞭 HeLa 細胞生長增殖( P =0.010、0.036);鑒定瞭 MAPK 通路的 KRAS 和 MKK 4( JNKK 1/MAP 2 K 4)是 miRNA-199 a 的靶基因,同時 KRAS 也是 miRNA-214的靶基因;miRNA-199 a 能夠抑製 HeLa 細胞 KRAS 和 MKK 4的錶達( P =0.231、0.011),而 miRNA-214也能夠抑製 KRAS 的錶達( P =0.029)。結論miRNA-199 a 和 miRNA-214通過調節 MAPK 信號途徑因子的錶達,參與瞭宮頸癌細胞的生長增殖。
목적:연구 miRNA-199 a 화 miRNA-214조공궁경암세포생장증식적분자궤제。방법용세포생장곡선실험화극륭형성실험연구 miRNA-199 a 화 miRNA-214대궁경암세포계 HeLa 세포생장증식적억제작용;이용신식학수단급쌍형광소매보고기인방법사선감정 miRNA-199 a 화 miRNA-214적파기인,용 Real-time PCR 화 Western 인적방법연구타문대파기인표체량적영향。결과 miRNA-199 a 화 miRNA-214현저억제료 HeLa 세포생장증식( P =0.010、0.036);감정료 MAPK 통로적 KRAS 화 MKK 4( JNKK 1/MAP 2 K 4)시 miRNA-199 a 적파기인,동시 KRAS 야시 miRNA-214적파기인;miRNA-199 a 능구억제 HeLa 세포 KRAS 화 MKK 4적표체( P =0.231、0.011),이 miRNA-214야능구억제 KRAS 적표체( P =0.029)。결론miRNA-199 a 화 miRNA-214통과조절 MAPK 신호도경인자적표체,삼여료궁경암세포적생장증식。
Objective To investigate the mechanism of regulations of miRNA-1 9 9 a and miRNA-2 1 4 in cell growth and proliferation of cervical cancer .Methods We used cell growth curves and cell clones formation assays to study effects of miRNA-1 9 9 a and miRNA-2 1 4 on growth of HeLa cells; Bioin-formatics tools and dual-luciferase report gene assays were used to screen and identify target genes of miRNA-1 9 9 a and miRNA-2 1 4 in MAPK pathway; Realtime-PCR and Western blotting were used to in-vestigate the inhibitory effects on those target genes of miRNA-1 9 9 a and miRNA-2 1 4 . Results We proved that miRNA-1 9 9 a , miRNA-2 1 4 could inhibit growth and proliferation of HeLa cells ( P =0.0 1 0 , 0.0 3 6 ); we identified miRNA-1 9 9 a ’ s target genes KRAS and MKK 4 , miRNA-2 1 4 ’ s target gene KRAS , and proved the inhibitory effects of miRNA-1 9 9 a and/or miRNA-2 1 4 on expression of those target genes ( P =0.2 3 1 , 0.0 1 1 , 0.0 2 9 ) .Conclusion miRNA-1 9 9 a , miRNA-2 1 4 could play im-portant roles in cell growth and proliferation of cervical cancer through affecting MAPK pathway .