长链非编码RNA LINC00467在卵巢癌的表达及其临床意义

The study of the expression of long non-coding RNA LINC00467 in ovarian cancer and its clinical significance

目的通过检测卵巢癌(ovarian cancer,OC)组织和卵巢癌细胞系中长链非编码RNA LINC00467的表达情况,探讨其与卵巢癌临床病理特征的关系及生物学特性,初步探讨LINC00467可能的调控机制。方法首先基于癌症基因组图谱(TCGA)的基因表达谱交互分析数据库,通过生物信息学分析评估LINC00467在卵巢癌组织中的表达水平,使用Starbase数据库预测LINC00467可能的miRNA作用靶点。再采用定量实时PCR(qPCR)检测66对卵巢癌组织和正常卵巢组织中LINC00467和miR-302a-3p的表达情况,分析其与卵巢癌临床病理特征的关系。进一步通过基因沉默技术,降低LINC00467的表达后,再采用CCK-8实验、集落形成实验、Transwell迁移和侵袭实验测定LINC00467对卵巢癌细胞的生物学特性的影响;使用双荧光素酶报告基因检测LINC00467和miR-302a-3p的结合关系,降低LINC00467的表达后检测miR-302a-3p的表达量。结果生物信息学分析提示癌症基因组图谱(TCGA)数据库中卵巢癌组织LINC00467表达明显高于正常对照组;实时定量PCR(qPCR)提示66例OC组LINC00467的表达明显高于正常卵巢组,差异有统计学意义(P<0.001),卵巢癌细胞系中LINC00467的表达亦明显高于人卵巢上皮细胞系(HOSEpiC),差异有统计学意义(P<0.001),验证了生物信息学分析揭示的结果。LINC00467高表达组和低表达组卵巢癌的病理分级、淋巴结转移情况以及FIGO分期差异有统计学意义(P<0.05)。生存分析结果表明低表达LINC00467的卵巢癌患者总生存期优于高表达LINC00467的卵巢癌患者,差异有统计学意义(P<0.01)。沉默LINC00467表达后,卵巢癌细胞的增殖活性、集落形成能力、迁移和侵袭能力都受到明显抑制,反而miR-302a-3p的表达量明显提高,差异有统计学意义(P<0.01),LINC00467与miR-302a-3p可以直接结合。结论长链非编码RNA LINC00467在卵巢癌组织及细胞中高表达,与卵巢癌细胞的增殖、转移能力相关,与卵巢癌患者预后不良呈正相关,其作用机制可能是LINC00467靶向调控miR-302a-3p而达到抑制作用。

Objective By detecting the expression of long-chain non-coding RNA LINC00467 in ovarian cancer tissues and ovarian cancer cell lines,to preliminarily explore its relationship with the clinicopathological characteristics of ovarian cancer and its biological characteristics in ovarian cancer,and the possible regulatory mechanism of LINC00467 was preliminarily explored.Methods Firstly,based on the gene expression profile interactive analysis database of Cancer Genome Atlas(TCGA),the expression level of LINC00467 in ovarian cancer tissue(Ovarian cancer,OC)was evaluated through bioinformatics analysis;then the possible miRNA targets of LINC00467 were predicted using the Starbase database.Then quantitative real-time PCR(q PCR)was used to detect the expression of LINC00467 in 66 pairs of ovarian cancer tissues and normal ovarian tissues,and analyze its relationship with the clinicopathological characteristics of ovarian cancer.Furthermore,after reducing the expression of LINC00467 through gene silencing technology,CCK-8 experiment,colony formation experiment,Transwell migration and invasion experiment were used to determine the effect of LINC00467 on the biological characteristics of ovarian cancer cells.The binding relationship between LINC00467 and miR-302a-3p was detected by dual-luciferase reporter gene,and the expression of miR-302a-3p was detected after reducing the expression of LINC00467.Results The bioinformatics analysis showed that the expression of LINC00467 in ovarian cancer tissue(OC)in the Cancer Genome Atlas(TCGA)database was significantly higher than that of the normal control group;quantitative real-time PCR(qPCR)indicated that the expression of LINC00467and miR-302a-3p in 66 cases of OC group was significantly higher than that of the normal ovarian group.The expression of LINC00467in ovarian cancer cell lines was also significantly higher than that in human ovarian epithelial cell lines(HOSEpiC),which verified the results revealed by bioinformatics analysis.Further analysis showed that the expression level of LINC00467 was positively correlated with the pathological grade of ovarian cancer,lymph node metastasis and FIGO staging.The results of survival analysis showed that the overall survival of OC patients with low expression of LINC00467 was better than that of OC patients with high expression of LINC00467.After silencing the expression of LINC00467,the proliferation activity,colony forming ability,migration and invasion ability of ovarian cancer cells were significantly inhibited.Conclusion The long-chain non-coding RNA LINC00467 is highly expressed in ovarian cancer tissues and cells,which is related to the proliferation and metastasis of ovarian cancer cells,and positively related to the poor prognosis of patients with ovarian cancer.Its mechanism may be that LINC00467 targets and regulates miR-302a-3p.