摘要
目的通过检测肝癌细胞系中长链非编码RNA MALAT1(lncRNA MALAT1)的表达情况,以及其对肝癌细胞增殖,迁移与体外成瘤能力的影响,探讨其机制。方法采用荧光定量PCR技术(qRT-PCR)分析肝癌细胞系Huh-7,HepG2和正常肝细胞系L-O2中的lncRNA MALAT1的表达情况。将Huh-7细胞系分为MALAT1RNAi载体的细胞株Huh-7-siMALAT1组(si-MALAT1组)、转染无关干扰序列的Huh-7-NC组(si-NC组)以及空白对照组(Blank组)。采用CCK-8法测定细胞的增殖能力。细胞划痕及集落形成实验测定细胞的迁移能力与体外成瘤能力。Western blot测定PI3K/Akt信号传导通路上的关键靶点PI3K及Akt蛋白的表达情况。结果肝癌细胞系Huh-7,HepG2中的lncRNA MALAT1表达明显高于正常肝细胞系L-O2(P<0.05)。转染MALAT1RNAi后,Huh-7、HepG2的MALAT1表达下调(P<0.05)。CCK-8结果显示:转染48h,si-MALAT1组与Blank组的OD450nm值分别为(0.881±0.05)与(1.151±0.06)(P<0.05);转染72h,si-MALAT1组与Blank组的OD450nm值分别为(1.092±0.06)与(1.578±0.05)(P<0.05)。si-MALAT1组细胞划痕愈合率(31.420±6.235)%显著低于Blank组(63.477±4.324)%(P<0.05)。si-MALAT1组体外成瘤能力(22.753±6.382)%明显低于Blank组(52.141±5.320)%(P<0.05)。与Blank组相比,si-MALAT1组PI3K、Akt表达量下降(P<0.05)。结论lncRNA MALAT1在肝癌细胞系中处于高表达状态,敲低lncRNA MALAT1可以抑制肝癌细胞的增殖,迁移及体外成瘤能力。其机制可能与lncRNA MALAT1影响PI3K/Akt信号传导通路上关键靶点的表达有关。
Objective To investigate the expression of long non-coding RNA malat1(lncRNA MALAT1)in hepatocellular carcinoma(HCC)cell lines and its effect on the proliferation,migration and tumorigenicity of HCC cells in vitro.Methods The expression of lncRNA MALAT1 in Huh-7,HepG2 and L-O2 cell lines was analyzed by qRTPCR.Huh-7 cell lines were divided into three groups:Huh7-siMALAT1 group,Huh7-NC group and blank group.CCK-8 method was used to determine the proliferation of the cells.Cell scratch and colony assay were used to determine the migration and tumorigenicity of the cells in vitro.Western blot was used to detect the expression of Akt and PI3 Kin PI3 K/Akt signaling pathway.Results The expression of lncRNA MALAT1 in Huh-7 and HepG2 cells was significantly higher than that in L-O2 cells(P<0.05).After transfection with MALAT1 RNAi,the expression of MALAT1 in Huh-7 was down regulated(P<0.05).CCK8 results showed that after 48 hours of transfection,the OD450 nm values of si-MALAT1 group and Blank group were(0.881±0.05)and(1.151±0.06)respectively(P<0.05).After 72 hours of transfection,the OD450 nm values of siMALAT1 group and Blank group were(1.092±0.06)and(1.578±0.05)respectively(P<0.05).The wound healing rate(31.420±6.235)%of si-MALAT1 cells was significantly lower than the Blank group(63.477±4.324)%(P<0.05).The tumorigenicity of si-MALAT1 group in vitro(22.753±6.382)%was significantly lower than that in Blank group(52.141±5.320)%(P<0.05).Compared with Blank group,the expression of Akt and PI3 Kwere down regulated in si-MALAT1 group(P<0.05).Conclusion LncRNA MALAT1 is highly expressed in HCC cell lines.Knockdown of lncRNA MALAT1 can inhibit the proliferation,migration and tumorigenesis of HCC cells in vitro.The mechanism may be that lncRNA MALAT1 affects the expression of key targets in PI3 K/Akt signaling pathway.
作者
吴惧
李贺
岳坤
程楠
徐键
尹敏
尹家俊
余杰
WU Ju;LI He;YUE Kun(Department of Hepatobiliary La paroscopic Surgery,Zhongshan Hospi-tal,DaLian University,Dalian 116001,China)
出处
《中国实验诊断学》
2021年第4期564-569,共6页
Chinese Journal of Laboratory Diagnosis
关键词
长链非编码RNA
MALAT1
肝癌细胞
增殖
迁移
信号传导通路
long non-coding RNA MALAT1
hepatocellular carcinoma cells
proliferation
migration
signal transduction pathway