长链非编码RNA FER1L4在肾癌组织中的表达及其对肾癌细胞增殖侵袭和迁移的影响

雷坤阳; 谢文杰; 孙庭

doi:10.12354/j.issn.1000-8179.2021.20210842

长链非编码RNA FER1L4在肾癌组织中的表达及其对肾癌细胞增殖侵袭和迁移的影响

Expression of long non-coding RNA FER1L4 in renal cell carcinoma tissues and itseffect on the proliferation, invasion, and migration of renal cell carcinoma cells

摘要

摘要:
目的探讨长链非编码RNA（long non-coding RNA，LncRNA）FER1L4在肾癌组织中的表达及其在体外肾癌细胞增殖、侵袭和迁移中的作用。
方法通过GEPIA数据库分析523例患者的肾癌组织和其中100例患者的配对癌旁组织中的LncRNA FER1L4表达，及516例肾癌患者的组织标本与其生存期之间的关系。收集2020年1月至2021年1月65例于南昌大学第一附属医院行肾癌根治术患者的肾癌组织及癌旁组织标本，定量逆转录聚合酶链式反应（reverse transcription-quantitative polymerase chain reaction，RT-qPCR）检测患者的肾癌组织中的LncRNA FER1L4表达，并分析其与患者临床病理学特征的相关性。构建LncRNA FER1L4沉默肾癌786-O和OSRC2细胞株，利用CCK8、划痕、Transwell以及平板克隆实验检测LncRNA FER1L4对体外肾癌细胞增殖、侵袭及迁移的影响。Western blot检测上皮-间质转化（epithelial-mesenchymal transition，EMT）通路相关蛋白的表达变化。
结果 GEPIA数据库分析结果表明，LncRNA FER1L4在肾癌的表达明显高于其癌旁组织，并且其表达与生存期呈负相关（P<0.001）。RT-qPCR结果显示，65例患者的肾癌组织中的LncRNA FER1L4表达为8.63±1.79，明显高于癌旁组织的1.84±0.95，两者比较差异具有统计学意义（P<0.001）。肾癌组织中的LncRNA FER1L4高表达与患者肿瘤直径、肿瘤分期、淋巴结转移及远处转移显著相关（均P<0.05）。CCK8、划痕、Transwell以及平板克隆实验结果显示，沉默LncRNA FER1L4表达后，肾癌786-O和OSRC2细胞的增殖、迁移及侵袭能力均明显减弱（均P<0.05）。Western blot检测显示，肾癌786-O和OSRC2细胞中沉默LncRNA FER1L4，Vimentin和N-cadherin表达下调，E-cadherin表达上调（均P<0.05）。
结论肾癌中LncRNA FER1L4高表达，并且与患者的肿瘤直径、肿瘤分期、淋巴结转移及远处转移相关。LncRNA FER1L4促进肾癌进展的作用可能部分与EMT相关。

Abstract:
Objective To investigate the expression of the long non-coding RNA (lncRNA) FER1L4 in renal cell carcinoma tissues and its role in the proliferation, invasion, and migration of renal cell carcinoma cells in vitro.
Methods The expression level of lncRNA FER1L4 in 523 renal cancer tissues and 100 paracancerous tissues and the relationship between lncRNA FER1L4 expression level in tissue samples from 516 renal cancer patients and the prognoses were analyzed using the GEPIA database. Tumor tissue and paracancerous tissue samples were collected from 65 renal cell carcinoma patients under treatment at The First Affiliated Hospital of Nanchang University from January 2020 to January 2021. Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) was used to determine the expression level of LncRNA FER1L4 in renal cell carcinoma tissues and cells, and the correlation between LncRNA FER1L4 expression and clinicopathological characteristics of patients were analyzed. LncRNA FER1L4-silenced 786-O and OSRC2 renal cell carcinoma cell lines were established. The effects of lncRNA FER1L4 on the proliferation, invasion, and migration of these cell lines in vitro were determined using the CCK8, wound healing, Transwell, and colony formation assays. Changes in the expression levels of proteins related to epithelial-mesenchymal transition (EMT) were detected by Western blot.
Results The results of the GEPIA database analysis showed that lncRNA FER1L4 expression was significantly higher in the renal cell carcinoma than in the matched paracancerous tissues and that its expression negatively correlated with survival (P<0.001). RT-qPCR results showed that lncRNA FER1L4 expression in renal cell carcinoma tissues of 65 patients (8.63 ± 1.79) was significantly higher than that in the matched adjacent tissues (1.84±0.95; P<0.001). The high lncRNA FER1L4 expression in the renal cell carcinoma tissues significantly correlated with the tumor diameter, tumor stage, lymph node metastasis, and distant metastasis (all P<0.05). Results of the CCK8, wound healing, Transwell, and colony formation assays showed that the proliferation, migration, and invasion ability of the 786-O and OSRC2 cells were significantly reduced after LncRNA FER1L4 silencing (P<0.05). Western blot results showed that Vimentin and N-cadherin were down-regulated, while E-cadherin was up-regulated in the LncRNA FER1L4-silenced 786-O and OSRC2 cells (P<0.05).
Conclusions LncRNA FER1L4 was highly expressed in renal cell carcinoma, correlating with tumor diameter, tumor stage, lymphatic metastasis, and distant metastasis. The promoting effect of LncRNA FER1L4 on renal cell carcinoma progression might be partially related to EMT.

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