Effect of LncRNA OSER1-AS1 targeted regulation on miR-433-3p and its biological significance in hepatocellular carcinoma
扫码查看
点击上方二维码区域，可以放大扫码查看
原文链接
NETL
NSTL
万方数据
目的 研究肝细胞肝癌(HCC)中长链非编码RNA(LncRNA)氧化应激反应丝氨酸丰富1反义RNA1(OSER1-AS1)靶向调控微小RNA(miR)-433-3p的作用及生物学意义.方法 选择唐山市第九医院手术切除的HCC组织和癌旁组织,培养 HCC 细胞株 SMMC-7721、HepG2、MHCC97H 及正常肝细胞株 HL-7702.检测 LncRNA OSER1-AS1、miR-433-3p的表达水平,比较HCC组织与癌旁组织、HCC细胞株与正常肝细胞株中LncRNA OSER1-AS1、miR-433-3p表达水平的差异.对转染MHCC97H细胞进行分组,转染阴性对照(NC)-siRNA为si-NC组,转染LncRNA OSER1-AS1-siRNA 为 si-OSER1-AS1组,转染 LncRNA OSER1-AS1-siRNA 及 NC miR 为 si-OSER1-AS1+miR-NC 组,转染 LncR-NA OSER 1-AS 1-siRNA及miR-433-3p抑制物为si-OSER1-AS1+miR-433-3p抑制物组.检测细胞增殖活力、凋亡率、增殖细胞核抗原(PCNA)及裂解型caspase-3(cleaved caspase-3)的表达水平,采用双荧光素酶报告基因实验验证LncR-NA OSER1-AS1靶向miR-433-3p.结果 HCC组织中LncRNA OSER1-AS1的表达水平高于癌旁组织(1.77±0.34 vs 1.00±0.21)、miR-433-3p 的表达水平低于癌旁组织(0.65±0.09 vs 1.00±0.28)(P＜0.05)且 LncRNA OSER1-AS1 与miR-433-3p 呈负相关(r=-0.351,P＜0.05);HCC 细胞株中 LncRNA OSER 1-AS 1 的表达水平高于 HL-7702 细胞、miR-433-3p的表达水平低于HL-7702细胞(P＜0.05)且MHCC97H细胞中上述变化最显著.si-OSER1-AS1组MHCC97H 细胞中 LncRNA OSER1-AS1、PCNA 的表达水平及增殖活力低于 si-NC 组(0.37±0.05 vs 1.00±0.11,0.33± 0.05 vs 0.92±0.12,0.51±0.09 vs 1.03±0.12),miR-433-3p、cleaved caspase-3 的表达水平及凋亡率高于 si-NC 组(1.88±0.25 vs 1.00±0.09,0.96±0.11 vs 0.44±0.06,9.39％±1.15％vs 3.82％±0.55％)(P＜0.05);si-OSER1-AS1+miR-433-3p 抑制物组 MHCC97H 细胞中 cleaved caspase-3 表达水平及凋亡率低于 si-OSER1-AS1+miR-NC 组(0.27±0.05 vs 0.91±0.10,6.04％±0.77％ vs 11.32％±1.32％),PCNA 的表达水平及增殖活力高于 si-OSER1-AS1+miR-NC 组(0.94±0.12 vs 0.48±0.06,0.95±0.11 vs 0.34±0.05)(P＜0.05).结论 LncRNA OSER1-AS1 靶向 miR-433-3p调控HCC细胞的增殖和凋亡.
Objective To study the effect of long noncoding RNA(LncRNA)OSER1-AS1 targeted regulation on miR-433-3p and its biological significance in hepatocellular carcinoma(HCC).Methods The HCC tissues and adjacent tissues were collect-ed,HCC cell lines SMMC-7721,HepG2,MHCC97H and normal hepatocyte line HL-7702 were cultured.The expression lev-els of LncRNA OSER1-AS1 and miR-433-3p were detected.The expression levels of LncRNA OSER1-AS1 and miR-433-3p in HCC tissues and adjacent tissues,HCC cell lines and normal hepatocyte lines were compared.The transferred MHCC97H cell were divided into 4 groups:transfected with negative control(NC)-siRNA as si-NC group,transfected with LncR-NA OSER1-AS1-siRNA as si-OSER1-AS1 group,transfected with LncRNA OSER1-AS1-siRNA and NC miR as si-OSER1-AS1+miR-NC group,transfected with LncRNA OSER1-AS1-siRNA and miR-433-3p inhibitor as si-OSER1-AS1+miR-433-3p inhibitor group.The cell proliferation activity,apoptosis rate,expression levels of proliferating cell nuclear antigen(PCNA)and cleaved caspase-3 were detected.The dual luciferase reporter gene assay was used to validate LncRNA OSER1-AS1 target-ing miR-433-3p.Results The expression level of LncRNA OSER1-AS1 in HCC was higher than that in adjacent tissues(1.77± 0.34 vs 1.00±0.21),the expression level of miR-433-3p was lower than that in adjacent tissues(0.65±0.09 vs 1.00±0.28)(P＜0.05),and there was negative correlation between LncRNA OSER1-AS1 and miR-433-3p(r=-0.351,P＜0.05).The expression level of LncRNA OSER1-AS1 in HCC cell was higher than that in HL-7702 cell,the expression level of miR-433-3p was lower than that of HL-7702 cell(P＜0.05),and the above changes were most significant in MHCC97H cell.The expression level of LncRNA OSER1-AS1 and PCNA,the proliferation viability of MHCC97H cell in si-OSER1-AS1 group were lower than those in si-NC group(0.37± 0.05 vs 1.00±0.11,0.33±0.05 vs 0.92±0.12,0.51±0.09 vs 1.03±0.12),the expression level of miR-433-3p and cleaved caspase-3,apoptosis rate in si-OSER1-AS1 group were higher than those in si-NC group(1.88±0.25 vs 1.00±0.09,0.96±0.11 vs 0.44±0.06,9.39％±1.15％ vs 3.82％±0.55％)(P＜0.05).The expression level of cleaved caspase-3,the apoptosis rate of MHCC97H cell in si-OSER1-AS1+miR-3p inhibitor group were lower than those in si-OSER1-AS1+miR-NC group(0.27± 0.05 vs 0.91±0.10,6.04％±0.77％ vs 11.32％±1.32％),the expression level of PCNA and proliferation activity were higher than those in si-OSER1-AS1+miR-NC group(0.94±0.12 vs 0.48±0.06,0.95±0.11 vs 0.34±0.05)(P＜0.05).Conclusion It is demonstrated that LncRNA OSER1-AS1 regulates the proliferation and apoptosis of HCC cell by targeting miR-433-3p.
NETL
NSTL
万方数据
