Lanzhou University Second Hospital, Lanzhou, Gansu, China.
Department of Gastroenterology, The First People's Hospital of Lanzhou, Lanzhou, Gansu, China.
DNA Cell Biol. 2021 Oct;40(10):1278-1289. doi: 10.1089/dna.2021.0235. Epub 2021 Sep 24.
Long noncoding RNAs (lncRNAs) represent promising therapeutic targets associated with hepatocellular carcinoma (HCC). lncRNA VPS9D1 antisense RNA 1 () regulates colon and prostate cancer, but its relevance in HCC remains to be clarified. Using microarray data from the NCBI Gene Expression Omnibus (GEO) database (GSE65485) and The Cancer Genome Atlas (TCGA) database, expression in HCC and normal liver tissue sample HCC were compared. Relative lncRNA expression was also measured through real-time quantitative PCR (qPCR) in 80 pairs of HCC tumor and paracancerous tissues and in human HCC cell lines. knockdown was achieved by transfecting these HCC cells with a specific siRNA construct and the proliferation of these cells was quantified through cell proliferation assays and colony formation assays, while flow cytometry was employed to assess their cell cycle progression. The role of the lncRNA as a regulator of HCC tumorigenesis was also assessed by subcutaneously implanting BALB/c nude mice with HepG2 cells stably expressing either sh- or a control shRNA construct. Mechanistic analyses were additionally conducted by examining and expression through western blotting and qPCR. expression was significantly increased in HCC tissues in the analyzed databases and our independent tissue samples. Elevated expression was related to larger tumor size and more advanced tumor, node, metastasis (TNM) stage, and HCC patients expressing higher levels of this lncRNA exhibited poorer survival outcomes. Knocking down impaired the proliferative and colony formation activity of HepG2 cells while promoting their apoptotic death. Consistently, silencing suppressed HCC tumor growth Mechanistically, was able to bind to the HuR protein and thereby influence the stability and expression of the mRNA, thus impacting HCC cell proliferation. The /HuR/CDK4 signaling axis regulates HCC tumor cell oncogenic activity, highlighting this pathway as a promising therapeutic target.
长链非编码 RNA（lncRNA）是与肝细胞癌（HCC）相关的有前途的治疗靶点。lncRNA VPS9D1 反义 RNA 1（）调节结肠癌和前列腺癌，但它在 HCC 中的相关性尚待阐明。使用来自 NCBI 基因表达综合数据库（GEO）数据库（GSE65485）和癌症基因组图谱（TCGA）数据库的微阵列数据，比较了 HCC 和正常肝组织样本中 HCC 的表达。通过实时定量 PCR（qPCR）在 80 对 HCC 肿瘤和癌旁组织以及人 HCC 细胞系中测量相对 lncRNA 表达。通过用特定的 siRNA 构建体转染这些 HCC 细胞来实现下调，并通过细胞增殖测定和集落形成测定来定量这些细胞的增殖，同时通过流式细胞术评估它们的细胞周期进程。还通过将稳定表达 sh-或对照 shRNA 构建体的 HepG2 细胞皮下植入 BALB/c 裸鼠来评估作为 HCC 肿瘤发生调节剂的作用。通过 Western blot 和 qPCR 进一步进行了机制分析，以检查和表达。在分析的数据库和我们的独立组织样本中，HCC 组织中显着增加了表达。升高的表达与更大的肿瘤大小和更先进的肿瘤、淋巴结、转移（TNM）阶段有关，并且表达这种 lncRNA 水平较高的 HCC 患者的生存结果较差。敲低会损害 HepG2 细胞的增殖和集落形成活性，同时促进其凋亡死亡。一致地，沉默抑制 HCC 肿瘤生长。在机制上，能够与 HuR 蛋白结合，从而影响的 mRNA 稳定性和表达，从而影响 HCC 细胞增殖。/HuR/CDK4 信号轴调节 HCC 肿瘤细胞致癌活性，强调该途径是有前途的治疗靶点。
