长链非编码RNA GSTM3TV2通过调节微小RNA-597对肝癌细胞恶性细胞生物学的影响

Effects of long non-coding RNA GSTM3TV2 on malignant biological behaviors of hepatoma cells by regulating microRNA-597

目的探讨长链非编码RNA(lncRNA)GSTM3TV2对肝癌细胞增殖、侵袭和转移的影响,并探讨其与微小RNA(miR)-597的相互作用。方法选取郑州大学附属南阳市中心医院、南阳市宛城区第一人民医院和郑州大学第一附属医院2016年5月到2018年5月收集的126例肝癌和癌旁组织作为研究对象,采用荧光定量聚合酶链反应(PCR)分析癌旁和肝癌组织中GSTM3TV2和miR-597表达水平;在肝癌细胞系HepG2建立lncRNA对照组和GSTM3TV2 KD组细胞系,采用细胞计数试剂盒(CCK-8)、Transwell和体内异体移植瘤实验检测GSTM3TV2的体内外功能;采用生物信息学和双荧光素酶报告基因分析GSTM3TV2与miR-597的关系;将miRNA抑制剂转染HepG2细胞,分析其作用机制。计量数据比较采用t检验。结果肝癌组织中GSTM3TV2表达水平(2.96±0.34)明显高于癌旁组织(2.96±0.34),差异有统计学意义(t=4.819,P<0.05)。lncRNA对照组细胞48 h吸光度(A)值(2.17±0.22)明显高于GSTM3TV2 KD组(1.09±0.20),差异有统计学意义(t=3.429,P<0.05)。lncRNA对照组细胞侵袭数量[(154.82±13.83)个]明显高于GSTM3TV2 KD组[(87.29±10.43)个],差异有统计学意义(t=5.018,P<0.05)。lncRNA对照组细胞体内成瘤体积[(1048.45±251.30)mm3]明显大于GSTM3TV2 KD组[(759.49±142.09)mm3],差异有统计学意义(t=4.719,P<0.05)。lncRNA对照组细胞体内肿瘤重量[(1.09±0.21)g]明显高于GSTM3TV2 KD组[(0.61±0.15)g],差异有统计学意义(t=3.185,P<0.05)。lncRNA对照组细胞淋巴结转移数量[(15.29±3.39)个]明显多于GSTM3TV2 KD组[(8.22±2.09)个],差异有统计学意义(t=3.103,P<0.05)。生物信息学和双荧光素酶活性结果显示miR-597是GSTM3TV2的靶基因。lncRNA对照组细胞miR-597表达水平(1.09±0.21)明显低于GSTM3TV2 KD组(2.80±0.26),差异有统计学意义(t=3.529,P<0.05)。结论lncRNA GSTM3TV2通过调节miR-597促进肝癌细胞增殖、侵袭和转移等恶性细胞生物学行为。

Objective To investigate the effect of long non-coding RNA(lncRNA)GSTM3TV2 on proliferation,invasion and metastasis of hepatoma cells and its interaction with microRNA(miR)-597.Methods A total of 126 cases of liver cancer and adjacent tissues collected in Nanyang Central Hospital Affiliated to Zhengzhou University,the Nanyang First People’s Hospital and the First Affiliated Hospital of Zhengzhou University from May 2016 to May 2018 were selected as the research objects.The expression levels of lncRNA GSTM3TV2 and miR-597 in adjacent and liver cancer tissues were analyzed by fluorescence quantitative polymerase chain reaction.The cell lines of lncRNA control group and GSTM3TV2 KD group were established in hepatoma cell line HepG2.The functions of GSTM3TV2 in vivo and in vitro were detected by cell counting kit-8(CCK-8),Transwell and allogeneic tumor transplantation in vivo.The relationship between GSTM3TV2 and miR-597 was analyzed by bioinformatics and double luciferase reporter gene.The measurement data were compared by t-test.Results The expression level of GSTM3TV2 in hepatocellular carcinoma(2.96±0.34)was significantly higher than that in adjacent tissues(2.96±0.34,t=4.819,P<0.05).The absorbance(A)value at 48 h in lncRNA control group(2.17±0.22)was significantly higher than that in GSTM3TV2 KD group(1.09±0.20,t=3.429,P<0.05).The number of invasive cells in lncRNA control group(154.82±13.83)was significantly greater than that in GSTM3TV2 KD group(87.29±10.43,t=5.018,P<0.05).The tumor volume in lncRNA control group[(1048.45±251.30)mm3]was significantly larger than that in GSTM3TV2 KD group[(759.49±142.09)mm3,t=4.719,P<0.05].The tumor weight in lncRNA control group[(1.09±0.21)g]was significantly higher than that in GSTM3TV2 KD group[(0.61±0.15)g,t=3.185,P<0.05].The number of lymph node metastases in lncRNA control group[(15.29±3.39)]was significantly greater than that in GSTM3TV2 KD group[(8.22±2.09),t=3.103,P<0.05].Bioinformatics and double luciferase activity showed that miR-597 was the target gene of GSTM3TV2.The expression level of miR-597 in lncRNA control group(1.09±0.21)was significantly lower than that in GSTM3TV2 KD group(2.80±0.26,t=3.529,P<0.05).Conclusion lncRNA GSTM3TV2 promotes the biological behaviors of malignant cells such as proliferation,invasion and metastasis of hepatoma cells by regulating miR-597.