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长链非编码 RNA TP73-AS1 通过调控 microRNA-103 促进肝癌的恶性进展。

LncRNA TP73-AS1 promotes malignant progression of hepatoma by regulating microRNA-103.

机构信息

Department of Medical, Xi'an Emergency Medical Center, Xi'an, China.

出版信息

Eur Rev Med Pharmacol Sci. 2019 Jun;23(11):4713-4722. doi: 10.26355/eurrev_201906_18052.

Abstract

OBJECTIVE

The aim of this study was to investigate long non-coding RNA (lncRNA) TP73-AS1 expression in hepatocellular carcinoma (HCC) tissues and cells, and to further investigate whether it can accelerate the progression of HCC by regulating microRNA-103.

PATIENTS AND METHODS

Quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) was performed to examine TP73-AS1 expression in 60 pairs of HCC tissues and adjacent ones, and the association between lncRNATP73-AS1 level and clinical indicators of HCC as well as patients' prognosis was analyzed. Meanwhile, qRT-PCR was used to further verify TP73-AS1 expression in HCC cell lines. The lncRNA TP73-AS1 knockdown model was constructed using lentivirus in the HCC cell lines, including Bel-7402 and HepG2. Cell counting kit-8 (CCK-8), 5-Ethynyl-2'-deoxyuridine (EdU), and flow cytometry assays were performed to figure out the influence of TP73-AS1 on the basic biological function of the HCC cells. Finally, the involved potential regulatory mechanism was explored using cell recovery experiments, and the relationship between TP73-AS1 and microRNA-103 was further studied.

RESULTS

QRT-PCR results indicated that TP73-AS1 expression in HCC samples was conspicuously enhanced compared with paracancerous tissues, and patients with a relatively high level of TP73-AS1 had a higher tumor stage and a lower overall survival rate. Meanwhile, the proliferation ability of cells in the sh-TP73-AS1 group was strikingly lower than that in the control group, while cell apoptosis showed the opposite trend. Besides, qRT-PCR results indicated a negative correlation between microRNA-103 and TP73-AS1 in HCC tissue specimens. The results of the luciferase reporting assay revealed that TP73-AS1 could be targeted by microRNA-103 through binding site. In addition, the cell recovery experiment demonstrated that TP73-AS1 and microRNA-103 might have a mutual regulation, and the two of which could together affect the malignant progression of HCC.

CONCLUSIONS

TP73-AS1 expression was conspicuously enhanced both in HCC tissues and cell lines, which were associated with advanced tumor stage and poor prognosis. In addition, TP73-AS1 could accelerate the proliferation of HCC cells by regulating microRNA-103.

摘要

目的

本研究旨在探讨长链非编码 RNA(lncRNA)TP73-AS1 在肝癌(HCC)组织和细胞中的表达,并进一步研究其是否可以通过调节 microRNA-103 加速 HCC 的进展。

方法

采用实时定量聚合酶链反应(qRT-PCR)检测 60 对 HCC 组织及其相邻组织中 TP73-AS1 的表达,分析 lncRNATP73-AS1 水平与 HCC 临床指标及患者预后的关系。同时,采用 qRT-PCR 进一步验证 HCC 细胞系中 TP73-AS1 的表达。采用慢病毒构建 HCC 细胞系 Bel-7402 和 HepG2 中的 lncRNA TP73-AS1 敲低模型。采用细胞计数试剂盒-8(CCK-8)、5-乙炔基-2'-脱氧尿苷(EdU)和流式细胞术检测 TP73-AS1 对 HCC 细胞基本生物学功能的影响。最后,通过细胞恢复实验探讨潜在的调控机制,并进一步研究 TP73-AS1 与 microRNA-103 的关系。

结果

qRT-PCR 结果表明,与癌旁组织相比,HCC 组织中 TP73-AS1 的表达明显升高,TP73-AS1 水平较高的患者肿瘤分期较高,总生存率较低。同时,sh-TP73-AS1 组细胞的增殖能力明显低于对照组,而细胞凋亡则呈现相反的趋势。此外,qRT-PCR 结果表明 HCC 组织标本中 microRNA-103 与 TP73-AS1 呈负相关。荧光素酶报告实验结果表明,TP73-AS1 可通过结合位点与 microRNA-103 靶向结合。此外,细胞恢复实验表明,TP73-AS1 和 microRNA-103 可能存在相互调节,两者可共同影响 HCC 的恶性进展。

结论

TP73-AS1 在 HCC 组织和细胞系中的表达明显增强,与肿瘤分期较晚和预后不良有关。此外,TP73-AS1 可通过调节 microRNA-103 加速 HCC 细胞的增殖。

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