长链非编码 RNA RPPH1 通过抑制胃癌中 P21 的表达来预测不良预后并调节细胞增殖和迁移。
LncRNA RPPH1 predicts poor prognosis and regulates cell proliferation and migration by repressing P21 expression in gastric cancer.
机构信息
Department of Clinical Laboratory, The Affiliated Hospital of Qingdao University, Qingdao, China.
出版信息
Eur Rev Med Pharmacol Sci. 2020 Nov;24(21):11072-11080. doi: 10.26355/eurrev_202011_23593.
OBJECTIVE
The purpose of this study was to explore the expression and biological functions of long non-coding ribonucleic acid (lncRNA) ribonuclease P RNA component H1 (RPPH1) in gastric cancer (GC), and to analyze the correlations of lncRNA expression with the clinical features and prognosis of GC patients.
PATIENTS AND METHODS
The relative expression of RPPH1 in tissue specimens from 60 GC patients was measured via quantitative Reverse Transcription-Polymerase Chain Reaction (qRT-PCR), and the correlations of RPPH1 expression with tumor-node-metastasis (TNM) stage, lymph node metastasis, etc. in GC patients were analyzed. Then, qRT-PCR was performed to detect the relative expression level of RPPH1 in GC cells. Moreover, colony formation assay, 5-Ethynyl-2'-deoxyuridine (EdU) staining, wound-healing assay, and transwell assay were employed to investigate the influence of RPPH1 on GC cell functions. After interfering in the expression of RPPH1, the changes in p21 (CDKN1A, cyclin dependent kinase inhibitor 1A) expression were determined through qRT-PCR and Western blotting.
RESULTS
It was shown in qRT-PCR assay results that the expression of RPPH1 was upregulated in 60 cases of GC tissues. Statistical analysis revealed that RPPH1 expression was positively correlated with the TNM stage, lymph node metastasis, and infiltration depth in GC patients. Besides, highly expressed lncRNA RPPH1 suggested poor prognosis of GC patients. Based on the results of qRT-PCR assay, the expression of RPPH1 in GC cells was upregulated. After interfering in RPPH1 expression, both colony formation assay and EdU staining indicated that the proliferative capacity of GC cells was repressed. Furthermore, it was manifested in the results of wound-healing and transwell assays that the migratory and invasive abilities of GC cells were weakened. Finally, the qRT-PCR and Western blotting assay results demonstrated that p21 expression was upregulated after interfering in the expression of RPPH1 in GC cells.
CONCLUSIONS
The expression of lncRNA RPPH1 is upregulated in GC, suggesting that the prognosis of the patients is poor. Highly expressed RPPH1 promotes the proliferation and metastasis of GC cells by regulating p21 expression.
目的
本研究旨在探讨长链非编码核糖核酸（lncRNA）核糖核酸酶 P RNA 成分 H1（RPPH1）在胃癌（GC）中的表达和生物学功能，并分析 lncRNA 表达与 GC 患者临床特征和预后的相关性。
方法
采用定量逆转录-聚合酶链反应（qRT-PCR）检测 60 例 GC 患者组织标本中 RPPH1 的相对表达水平，并分析 RPPH1 表达与 GC 患者肿瘤-淋巴结-转移（TNM）分期、淋巴结转移等的相关性。然后，采用 qRT-PCR 检测 GC 细胞中 RPPH1 的相对表达水平。此外，采用集落形成实验、5-乙炔基-2'-脱氧尿苷（EdU）染色、划痕愈合实验和 Transwell 实验研究 RPPH1 对 GC 细胞功能的影响。干扰 RPPH1 表达后，通过 qRT-PCR 和 Western blot 检测 p21（CDKN1A，细胞周期蛋白依赖性激酶抑制剂 1A）表达的变化。
结果
qRT-PCR 检测结果显示，60 例 GC 组织中 RPPH1 的表达上调。统计分析显示，RPPH1 表达与 GC 患者的 TNM 分期、淋巴结转移和浸润深度呈正相关。此外，高表达的 lncRNA RPPH1 提示 GC 患者预后不良。基于 qRT-PCR 检测结果，GC 细胞中 RPPH1 的表达上调。干扰 RPPH1 表达后，集落形成实验和 EdU 染色均表明 GC 细胞的增殖能力受到抑制。此外，划痕愈合和 Transwell 实验结果表明，GC 细胞的迁移和侵袭能力减弱。最后，qRT-PCR 和 Western blot 检测结果表明，干扰 GC 细胞中 RPPH1 的表达后，p21 的表达上调。
结论
lncRNA RPPH1 在 GC 中表达上调，提示患者预后不良。高表达的 RPPH1 通过调节 p21 的表达促进 GC 细胞的增殖和转移。
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