| 摘要: |
| 目的 探讨lncRNA LINC00342在肾透明细胞癌（ccRCC）中的作用及其机制。方法 通过基因表达谱动态分析（GEPIA）数据库分析LINC00342在ccRCC组织中的表达及其表达水平与患者生存率的关系。采用qPCR检测ccRCC组织及ccRCC细胞系中LINC00342的表达，并分析其与ccRCC患者临床病理学特点的相关性。将ccRCC细胞分为siRNA组、阴性对照组和空白对照组，通过CCK-8实验、划痕愈合实验及Transwell实验分别验证LINC00342对ccRCC细胞增殖、迁移及侵袭的影响，采用蛋白质印迹法检测上皮间质转化（EMT）相关蛋白的表达变化。通过生物信息学方法预测LINC00342的靶分子，并采用双萤光素酶报告基因实验验证LINC00342与miRNA-384的靶向关系。结果 GEPIA数据库的分析结果表明，LINC00342在ccRCC组织中高表达，并且高表达的ccRCC患者总生存率较低（P均<0.05）。qPCR结果显示，ccRCC组织及ccRCC细胞中LINC00342的表达水平均升高（P均<0.05）。LINC00342高表达的患者肿瘤直径更大、临床分期更晚（P均<0.05）。通过siRNA沉默LINC00342的表达能够抑制ccRCC细胞的增殖、迁移及侵袭，下调波形蛋白和神经钙黏素的表达，上调上皮钙黏素的表达（PP均<0.05）。生物信息学分析结果显示miRNA-384存在LINC00342的结合位点，双萤光素酶报告基因实验结果表明miRNA-384是LINC00342的下游靶分子之一。结论 LINC00342可以促进ccRCC细胞的恶性表型。miRNA-384是LINC00342的下游靶分子之一。 |
| 关键词: 肾肿瘤 透明细胞癌 长链非编码RNA LINC00342 微RNA-384 上皮间质转化 生物信息学 |
| DOI：10.16781/j.CN31-2187/R.20210991 |
| 投稿时间：2021-09-28修订日期：2021-12-06 |
| 基金项目:江西省卫生健康委科技计划（202210397）. |
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| Long non-coding RNA LINC00342 promotes proliferation, migration and invasion of clear cell renal cell carcinoma cells by targeting microRNA-384 |
| XIE Wen-jie,LEI Kun-yang,SUN Ting* |
(Department of Urology, The First Affiliated Hospital of Nanchang University, Nanchang 330006, Jiangxi, China
*Corresponding author) |
| Abstract: |
| Objective To investigate the role of long non-coding RNA (lncRNA) LINC00342 in clear cell renal cell carcinoma (ccRCC) and its mechanism. Methods The expression of LINC00342 in ccRCC tissues was analyzed by the Gene Expression Profiling Interactive Analysis (GEPIA) database, and the relationship between LINC00342 expression and patient survival was analyzed. The expression of LINC00342 in ccRCC tissues and ccRCC cell lines was detected by quantitative polymerase chain reaction (qPCR), and its correlation with the clinicopathological characteristics of patients was analyzed. The ccRCC cells were divided into small interfering RNA group, negative control group, and blank group. The effects of LINC00342 on the proliferation, migration and invasion of ccRCC cells were verified by cell counting kit 8 (CCK-8) assay, scratch wound healing assay, and Transwell invasion assay. The expression of proteins involved in the epithelialmesenchymal transition (EMT) pathway was detected by Western blotting. The target genes of LINC00342 were predicted by bioinformatics, and the targeting relationship of LINC00342 and microRNA (miRNA)-384 was validated by dual-luciferase reporter gene assay. Results The analysis of the GEPIA database showed that LINC00342 was highly expressed in ccRCC tissues, and the patients with high LINC00342 expression had a low overall survival rate (both P<0.05). The qPCR results showed that the expression of LINC00342 was significantly increased in both ccRCC tissues and ccRCC cells (both P<0.05). The patients with high LINC00342 expression had larger tumor size and later clinical stage (both P<0.05). Silencing the expression of LINC00342 by small interfering RNA inhibited the proliferation, migration and invasion of ccRCC cells (all P< 0.05), down-regulated the expression of vimentin and N-cadherin, and up-regulated the expression of E-cadherin (all P<0.05). Bioinformatic analysis showed that miRNA-384 had a binding site with LINC00342, and the results of dual-luciferase reporter gene assay indicated that miRNA-384 was one of the downstream target genes of LINC00342. Conclusion LINC00342 can promote the malignant phenotype of ccRCC cells. miRNA-384 is one of the downstream target genes of LINC00342. |
| Key words: kidney neoplasms clear cell carcinoma long non-coding RNA LINC00342 microRNA-384 epithelialmesenchymal transition bioinformatics |
