长链非编码RNA LOC100288637在原发性肝癌中差异性表达的生物学意义及相关机制研究

Research on the biological significance of the LncRNA LOC100288637 expression differences in primary hepatic carcinoma and the related mechanism

目的研究长链非编码RNA(LncRNA)LOC100288637在肝癌中的表达差异,对肝癌细胞增殖的影响及相关机制。方法分析GEO数据集GSE58043和GSE10694,得出LOC100288637及hsa-miR-101-3p在肝癌组织中差异表达,RNA-hybrid分析LOC100288637与hsa-miR-101-3p的结合具有可能性。逆转录-聚合酶链式反应在组织和细胞水平检测LOC100288637表达量。荧光原位杂交观察LOC100288637在细胞中的定位。敲低LOC100288637后CCK-8检测细胞增殖情况。过表达hsamiR-101-3p后,检测LOC100288637的变化。结果LOC100288637在肝癌组织中的表达量高于癌旁组织(P<0.05);LOC100288637在肝癌细胞系中的表达量高于肝细胞系(P<0.05)。LOC100288637在HepG2细胞核质均有表达,以细胞质居多。敲低LOC100288637后HepG2细胞增殖活性降低(P<0.01)。过表达hsa-miR-101-3p后,LOC100288637表达量下降(P<0.05)。结论 LncRNA LOC100288637可能在肝癌发生,发展过程中起重要作用并接受hsa-miR-101-3p靶向调控影响肝癌细胞的增殖。

Objective To explore the expression differences of long noncoding RNA（LncRNA） LOC100288637 in liver cancer,the effect of LOC100288637 on liver cancer cell proliferation and the relevant mechanisms. Methods Based on the analysis of the GEO database data sets GSE58043 and GSE10694,we found that both LOC100288637 and hsa-miR-101-3p has obvious expression differences in liver cancer tissues. RNA hybrid revealed the possibility of combination between LOC100288637 and hsa-miR-101-3p; RT-PCR was performed to measure the expression level of LOC100288637 in tissues and cells; fluorescence in situ hybridization was used to observe LOC100288637 localization in cells;cell proliferation was determined by CCK8 experiment after LOC100288637 siRNA knock down. The expression of LOC100288637 in cells were measured after treated with hsa-miR-101-3p mimics. Results Relative quantitative expression of LOC100288637 in liver cancer tissues group was significantly higher than that in no tumor tissues group （P（0.05）;relative quantitative expression of LOC100288637 in liver cancer cell lines were significantly higher than that in normal liver cell line（P（0.05）. LOC100288637 was located in both cytoplasm and nucleus of HepG2 cells,and mainly in cytoplasm. Cell proliferation vitality of HepG2 reduced after treated with LOC100288637-siRNA（P〈0.01）. Relative quantitative expression of LOC100288637 in HepG2 reduced after treated with hsa-miR-101-3p mimics （P （0. 05）. Conclusion LncRNA LOC100288637 may play an important role in liver cancer development,it can be down regulated by hsa-miR-101-3p and affect the proliferation of liver cancer cells.