CAJ | 학술논문

目的研究全反式维甲酸(all-trans-retinoicacid,ATRA)对肝癌细胞增殖的影响并探讨可能的作用机制.方法利用ATRA处理肝癌细胞系HepG2与SMMC-7721,选择MTT法分析细胞增殖.利用实时PCR方法研究ATRA对miR-18a表达的影响,并选择特异抑制剂Anti-miR-18a处理肝癌细胞,利用MTT法分析细胞增殖.设计拯救实验,利用ATRA处理转染miR-18a模拟物的肝癌细胞系,利用MTT法分析细胞增殖情况.结果 ATRA可以有效抑制肝癌细胞系HepG2和SMMC-7721的增殖,A490吸光度值分析显示,HepG2经ATRA处理后,细胞增殖分别被抑制74％(P＜0.05,36 h)、72％(P＜0.01,48 h)、67％(P＜0.05,72 h)；SMMC-7721经ATRA处理后,细胞增殖分别被抑制68％ (P＜0.05,48 h)、64％ (P＜0.01,72 h).miR-18a在肝癌细胞HepG2与SMMC-7721中的表达水平分别上调4.7倍(P＜0.05)、3.8倍(P＜0.05); ATRA处理后,HepG2与SMMC-7721内源性miR-18a的表达水平分别下调67％(P＜0.05)与56％(P＜0.05).过表达miR-18a的肝癌细胞HepG2与SMMC-7721对ATRA的抑制作用出现耐受,其中HepG2细胞增殖上调1.2倍(P＜0.05),SMMC-7721细胞增殖上调1.25倍(P＜0.05,24 h)与1.2倍(P＜0.05,48 h).结论 ATRA可通过下调肝癌细胞内源性miR-18a的表达,抑制细胞增殖.
목적 연구전반식유갑산(all-trans-retinoicacid,ATRA)대간암세포증식적영향병탐토가능적작용궤제.방법 이용ATRA처리간암세포계HepG2여SMMC-7721,선택MTT법분석세포증식.이용실시PCR방법연구ATRA대miR-18a표체적영향,병선택특이억제제Anti-miR-18a처리간암세포,이용MTT법분석세포증식.설계증구실험,이용ATRA처리전염miR-18a모의물적간암세포계,이용MTT법분석세포증식정황.결과 ATRA가이유효억제간암세포계HepG2화SMMC-7721적증식,A490흡광도치분석현시,HepG2경ATRA처리후,세포증식분별피억제74％(P＜0.05,36 h)、72％(P＜0.01,48 h)、67％(P＜0.05,72 h)；SMMC-7721경ATRA처리후,세포증식분별피억제68％ (P＜0.05,48 h)、64％ (P＜0.01,72 h).miR-18a재간암세포HepG2여SMMC-7721중적표체수평분별상조4.7배(P＜0.05)、3.8배(P＜0.05); ATRA처리후,HepG2여SMMC-7721내원성miR-18a적표체수평분별하조67％(P＜0.05)여56％(P＜0.05).과표체miR-18a적간암세포HepG2여SMMC-7721대ATRA적억제작용출현내수,기중HepG2세포증식상조1.2배(P＜0.05),SMMC-7721세포증식상조1.25배(P＜0.05,24 h)여1.2배(P＜0.05,48 h).결론 ATRA가통과하조간암세포내원성miR-18a적표체,억제세포증식.
Objective To investigate the inhibitory effect of all-tram-retinoicacid (ATRA) on HCC cell growth and probe the potential molecular mechanism.Methods HCC cell lines,HepG2 and SMMC-7721 were treated by ATRA and cell growth was analyzed by using MTT assay.The expression levels of miR-18a were evaluated in HepG2 and SMMC-7721,compared with the normal livers pool by using RealTime PCR analysis.Cell growth analysis by using MTT assay was performed on HepG2 and SMMC-7721 after transfection with anti-miR-18a.Rescued assay was designed to probe the mechanism of ATRA on cell growth by using ATRA with or without miR-18a mimic.Results HepG2 cell growth was suppressed about 74％ (P＜0.05,36 h),72％ (P＜0.01,48 h),and 67％ (P＜0.05,72 h) and SMMC-7721 cell growth was inhibited about 68％ (P＜0.05,48 h),and 64％ (P＜0.05,72 h) after treatment with ATRA,compared with the cells treated with Ethanol.MiR-18a expression was up-regulated in HepG2 and SMMC-7721 cell lines about 4.7- and 3.8-fold (P＜0.05),respectively.Endogenous miR-18a levels were down-regulated by ATRA about 67％ and 56％ (P＜0.05).The inhibitory effect of ATRA on HCC cell growth was reversed about 1.2-fold (P＜0.05,48 h) by overexpression of miR-18a in HepG2 cells and cell growth of SMMC-7721 was enhanced about 1.25- and 1.2-fold (P＜0.05,24 and 48 h) with ectopic expression of miR-18a.Conclusion HCC cells growth is suppressed by ATRA through miR-18a mediated network.