长链非编码RNA AURKAPS1调控肝癌细胞增殖、运动迁移能力的研究

Study on the ability of long non-coding RNA AURKAPS1 to regulate the proliferation,movement and migration of hepatocellular carcinoma cells

目的探讨长链非编码RNA(lncRNA)AURKAPS1对肝癌细胞增殖、运动迁移能力的影响。方法采用慢病毒包被质粒感染肝癌细胞构建AURKAPS1过表达细胞系,实时荧光定量PCR检测慢病毒感染后的AURKAPS1的表达量。将肝癌细胞系随机分为对照组,AURKAPS1组(对照组无任何干预,AURKAPS1组仅接受AURKAPS1干预);采用细胞计数试剂盒(CCK-8)细胞增殖实验、克隆形成和流式细胞分选方法体外检测过表达AURKAPS1对肝癌细胞增殖能力的影响;采用皮下注射裸鼠荷瘤实验体内检测过表达AURKAPS1对肝癌细胞增殖能力的影响。划痕实验体外检测过表达AURKAPS1对肝癌细胞运动能力的影响。所得数据组间采用独立样本t检验及单因素方差分析,多组间采用重复测量方差分析、LSD检验,以P<0.05为差异有统计学意义。结果过表达AURKAPS1后,肝癌细胞HepG2与BEL-7402细胞增殖都有增强(F_(HepG2)=205.593,P<0.001,η^(2)=0.934,F_(BEL-7402)=161.641,P<0.001,η^(2)=0.976);过表达AURKAPS1后,肝癌细胞HepG2和BEL-7402的克隆数均高于对照组(t_(BEL7402)=-9.806,P<0.01;t_(HepG2)=-14.425,P<0.01),AURKAPS1组BEL-7402与HepG2的G2和S期细胞比例相较于对照组明显高于G1期(F_(BEL-7402)=93.091,P<0.01,F_(HepG2)=360.071,P<0.001);裸鼠荷瘤结果显示肝癌细胞AURKAPS1组重量高于对照组(t=-2.427,P<0.05);AURKAPS1组HepG2和BEL-7402细胞平均划痕距离低于对照组(F_(HepG2)=125.472,P<0.001;F_(BEL-7402)=4465.901,P<0.001)。结论过表达AURKAPS1促进肝癌细胞生长增殖、促进肝癌细胞周期从G_(1)期向G_(2)和S期转变;增强肝癌细胞运动、迁移的能力。

Objective To investigate the effects of long non-coding RNA(lncRNA)AURKAPS1 on the proliferation,movement and migration of hepatoma cells.Methods The overexpression cell line of AURKAPS1 was constructed by infecting hepatoma cells with lentivirus coated plasmids.The expression of AURKAPS1 after lentivirus infection was detected by real-time fluorescent quantitative PCR.The hepatoma cell line was randomly divided into control group,AURKAPS1 group(control group had no intervention,AURKAPS1 group only received AURKAPS1 intervention);Cell counting kit-8(CCK-8)cell proliferation test,clone formation and flow cytometry were used to detect the effect of overexpression of AURKAPS1 on the proliferation of liver cancer cells in vitro;The effect of overexpression of AURKAPS1 on the proliferation of hepatoma cells was detected by subcutaneous injection of tumor bearing nude mice in vivo.Scratch test was used to detect the effect of overexpression of AURKAPS1 on the motility of liver cancer cells in vitro.The obtained data were tested by independent t-test among groups.Repeated measurement analysis of variance and LSD test were used among groups.The difference was statistically significant with P<0.05.Results After overexpression of AURKAPS1,the proliferation of HepG2 and BEL-7402 cells was enhanced(F_(HepG2)=205.593,P<0.001,η^(2)=0.934,F_(BEL-7402)=161.641,P<0.001,η^(2)=0.976);The clone numbers of HepG2 and BEL-7402 cells were higher than those of the control group(t_(BEL7402)=-9.806,P<0.01;t_(HepG2)=-14.425,P<0.01).The proportion of cells in G2 and S phases of BEL-7402 cells and HepG2 cells was significantly higher than that of the control group(F_(BEL-7402)=93.091,P<0.01,F_(HepG2)=360.071,P<0.001);The results of tumor bearing nude mice showed that the quality of AURKAPS1 group was higher than that of the control group(t=-2.427,P<0.05).The average scratch distance of HepG2 and BEL-7402 cells in AURKAPS1 group was lower than that in the control group(F_(HepG2)=125.472,P<0.001;F_(BEL-7402)=4465.01,P<0.001).Conclusion Overexpression of AURKAPS1 promotes the growth and proliferation of hepatoma cells,and promotes the transformation of hepatoma cell cycle from G_(1) phase to G_(2) and S phase;Enhance the ability of movement and migration of liver cancer cells.