长链非编码RNA CASC9通过下调miR-424-5p促进肝癌细胞的增殖、迁移并抑制其凋亡。

LncRNA CASC9 promotes proliferation, migration and inhibits apoptosis of hepatocellular carcinoma cells by down-regulating miR-424-5p.

作者信息

Yao Jingjing, Fu Jindong, Liu Ying, Qu Wei, Wang Guangdong, Yan Zaojun

机构信息

Department of Gastroenterology, People's Hospital of Rizhao, China.

Rizhao Health School, China.

出版信息

Ann Hepatol. 2021 Jul-Aug;23:100297. doi: 10.1016/j.aohep.2020.100297. Epub 2020 Dec 17.

Abstract

INTRODUCTION AND OBJECTIVES

CASC9 and miR-424-5p are closely related with hepatocellular carcinoma (HCC) progression. This study aimed to evaluate the effect of CASC9 involved with miR-424-5p on the development of HCC.

MATERIALS AND METHODS

qRT-PCR was performed to determine the mRNA expressions of CASC9 and miR-424-5p in HCC tissues/cells and adjacent normal tissues/human hepatic epithelial cells, and to analyze the relationship of CASC9 with the clinico-pathological characteristics and prognosis of HCC patients. Then, cell proliferation was measured by CCK-8 and clone formation assays. Apoptosis of HCC cells was measured by flow cytometry. Besides, cell migration and invasion were determined by scratch wound-healing and Transwell assays, respectively. DIANA-LncBase V2 and dual luciferase reporter gene assay were used to verify the targeted relationship between CASC9 and miR-424-5p. Bcl-2, Bax and cleaved caspase-3 expressions were detected by Western blot.

RESULTS

Higher expression of CASC9 was observed in HCC tissues/ cells than in adjacent normal tissues/ human hepatic epithelial cells, and was closely linked to poor prognosis of HCC, tumor size, TNM stage, differentiation degree, lymph node metastasis and alpha-fetoprotein (AFP). Down-regulation of CASC9 decreased the proliferation, invasion and migration of HCC cells while enhancing apoptosis. Besides, CASC9 was negatively correlated with miR-424-5p. MiR-424-5p inhibitor enhanced cell proliferation, invasion and migration while decreasing apoptosis. Interestingly, siRNA-CASC9 partially offset the effects of miR-424-5p inhibitor on HCC cells.

CONCLUSION

CASC9 promoted proliferation, invasion and migration and inhibited apoptosis in HCC cells by inhibiting miR-424-5p.

摘要

引言与目的

CASC9和miR - 424 - 5p与肝细胞癌(HCC)进展密切相关。本研究旨在评估CASC9与miR - 424 - 5p对HCC发生发展的影响。

材料与方法

采用qRT - PCR检测CASC9和miR - 424 - 5p在HCC组织/细胞及癌旁正常组织/人肝上皮细胞中的mRNA表达,并分析CASC9与HCC患者临床病理特征及预后的关系。然后,通过CCK - 8和克隆形成实验检测细胞增殖情况。采用流式细胞术检测HCC细胞凋亡情况。此外,分别通过划痕愈合实验和Transwell实验检测细胞迁移和侵袭能力。利用DIANA - LncBase V2和双荧光素酶报告基因实验验证CASC9与miR - 424 - 5p的靶向关系。通过蛋白质免疫印迹法检测Bcl - 2、Bax和裂解的caspase - 3表达。

结果

与癌旁正常组织/人肝上皮细胞相比,CASC9在HCC组织/细胞中表达更高,且与HCC预后不良、肿瘤大小、TNM分期、分化程度、淋巴结转移及甲胎蛋白(AFP)密切相关。下调CASC9可降低HCC细胞的增殖、侵袭和迁移能力,同时增强细胞凋亡。此外,CASC9与miR - 424 - 5p呈负相关。miR - 424 - 5p抑制剂可增强细胞增殖、侵袭和迁移能力,同时降低细胞凋亡。有趣的是,siRNA - CASC9可部分抵消miR - 用424 - 5p抑制剂对HCC细胞的影响。

结论

CASC9通过抑制miR - 424 - 5p促进HCC细胞增殖、侵袭和迁移并抑制细胞凋亡。

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