实用医学杂志 ›› 2022, Vol. 38 ›› Issue (12): 1486-1493.doi: 10.3969/j.issn.1006⁃5725.2022.12.009

• 基础研究 • 上一篇    下一篇

长链非编码RNA LINC00467在卵巢癌的表达及其临床意义

李剑琦 生秀杰   

  1. 广州医科大学附属第三医院妇科,广东省产科重大疾病重点实验室,广东省普通高校生殖与遗传重点实 验室(广州510150)

  • 出版日期:2022-06-25 发布日期:2022-06-25
  • 通讯作者: 生秀杰 E⁃mail:2008691150@gzhmu.edu.cn
  • 基金资助:
    广东省自然科学基金项目(编号:2020A1515010082)

The study of the expression of long non⁃coding RNA LINC00467 in ovarian cancer and its clinical signifi⁃ cance

LI Jianqi,SHENG Xiujie.   

  1. Department of Obstetrics and Gynecology,the Third Affiliated Hospital of Guang⁃ zhou Medical University,Key Laboratory of Major Obstetric Diseases of Guangdong Province,Key Laboratory of Re⁃ production and Genetic of Guangdong Higher Education Institutes,Guangzhou 510150,China

  • Online:2022-06-25 Published:2022-06-25
  • Contact: SHENG Xiujie E⁃mail:2008691150@gzhmu.edu.cn ​

摘要:

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 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 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。

关键词:

长链非编码RNA, LINC00467, 卵巢癌, 生存分析

Abstract:

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(qPCR)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 expres⁃ sion 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⁃lucif⁃erase 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.

Key words:

long non?coding RNA, LINC00467, ovarian cancer, survival analysis