摘要
背景与目的:缺氧是肝细胞癌(HCC)微环境的关键特征之一。缺氧可诱导编码基因和非编码RNA的表达,从而影响HCC的进展。既往研究发现缺氧反应性长链非编码RNA(lncRNA)AC114803与肾细胞癌预后相关,但其在HCC中尚未见研究。因此,本研究探讨HCC中lncRNA-AC114803表达与缺氧的关系及其功能。方法:低氧(1%O2)诱导后,用lncRNA芯片检测Hep3B细胞中缺氧调控的lncRNA,并用q RT-PCR在HCCLM3和Hep3B细胞中验证。通过TCGA数据分析AC114803在HCC组织中的表达及其与患者预后的关系。用AC114803过表达质粒转染HCCLM3和Hep3B细胞后,分别用CCK-8、细胞划痕实验检测细胞增殖和迁移能力的变化,用Western blot检测相关蛋白的表达变化。结果:lncRNA芯片分析结果显示,AC114803是Hep3B细胞中在缺氧环境下明显变化的lncRNA之一;q RT-PCR验证结果显示,HCCLM3和Hep3B细胞中仅AC114803的表达在缺氧环境下明显上调(均P<0.05)。TCGA数据分析显示,AC114803在HCC组织表达明显高于正常肝组织,且AC114803高表达患者生存率明显低于AC114803低表达患者(均P<0.05)。HCC细胞(HCCLM3、Hep3B)过表达AC114803后,增殖和迁移能力明显增强,间质标志物N-cadherin和vimentin以及抗凋亡蛋白bcl-2和细胞周期蛋白CCND1明显上调,而上皮标志物E-cadherin和促凋亡蛋白bax明显下调(均P<0.05)。结论:缺氧可诱导HCC细胞中AC114803的表达,其高表达与HCC患者的不良预后密切相关,AC114803可能通过调控上皮间质转化从而促进细胞增殖、迁移。
Background and Aims: Hypoxia is one of the key features of microenvironment of hepatocellular carcinoma(HCC). Hypoxia can induce the expressions of coding genes and non-coding RNAs thus,affecting the progression of HCC. Previous studies demonstrated that the hypoxia responsive long noncoding RNA(lncRNA) AC114803 is related to the prognosis of renal cell carcinoma. However, its role in HCC has not been studied. Therefore, this study was performed to investigate the relationship between AC114803 expression and hypoxia and its function in HCC cells.Methods: After hypoxia exposure, the hypoxia-regulated lncRNAs in Hep3 B cells were detected by lncRNA microarray, and then were verified in HCCLM3 and Hep3 B cells by q RT-PCR. The expression of AC114803 in HCC tissues and its relationship with the prognosis of patients were analyzed in TCGA samples. In HCCLM3 and Hep3 B cells after transfection with AC114803 overexpression plasmids, the changes in proliferative and migration abilities were determined by CCK-8 assay and wound healing assay, and changes in expressions of the relevant proteins were measured by Western blot analysis.Results: LncRNA microarray analysis showed AC114803 was one of the significantly changed lncRNAs in Hep3 B cells under hypoxia condition, and q RT-PCR verification showed that only the expression in AC114803 was significantly up-regulated in either HCCLM3 cells or Hep3 B cells(all P<0.05). TCGA data analysis showed that the expression of AC114803 in HCC tissues were significantly higher than that in normal liver tissues, and the survival rate in patients with high AC114803 expression was significantly lower than in those with low AC114803 expression(both P<0.05). After AC114803 overexpression, the proliferation and migration abilities of HCC cells(HCCLM3 and Hep3 B) were significantly enhanced, and the expressions of the mesenchymal markers N-cadherin and vimentin as well as the anti-apoptotic protein bcl-2 and CCND1 were significantly up-regulated, while the epithelial marker E-cadherin and the pro-apoptotic protein bax were significantly down-regulated(all P<0.05).Conclusion: Hypoxia can induce the expression of AC114803 in HCC cells, and high expression of AC114803 is closely associated with the poor prognosis of HCC patients. The action of AC114803 may probably be associated with promoting cell proliferation and migration by regulating the epithelialmesenchymal transition process.
作者
肖雪莲
刘润坤
涂康生
XIAO Xuelian;LIU Runkun;TU Kangsheng(Department of Hepatobiliary Surgery,the First Affiliated Hospital of Xi'an Jiaotong University,Xi'an 710000,China)
出处
《中国普通外科杂志》
CAS
CSCD
北大核心
2022年第1期89-96,共8页
China Journal of General Surgery
基金
陕西省科技厅自然科学基础研究计划基金资助项目(2020JC-36)。
关键词
癌
肝细胞
低氧
RNA
长链非编码
预后
肿瘤浸润
Carcinoma
Hepatocellular
Hypoxia
RNA
Long Noncoding
Prognosis
Neoplasm Invasiveness
