Long noncoding RNAs (lncRNAs) represent promising therapeutic targets associated with hepatocellular carcinoma (HCC). lncRNA VPS9D1 antisense RNA 1 (VPS9D1-AS1) 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, VPS9D1-AS1 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. VPS9D1-AS1 knockdown was achieved by transfecting these HCC cells with a specific siRNA construct in vitro, 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 VPS9D1-AS1 lncRNA as a regulator of HCC tumorigenesis was also assessed in vivo by subcutaneously implanting BALB/c nude mice with HepG2 cells stably expressing either sh-VPS9D1-AS1 or a control shRNA construct. Mechanistic analyses were additionally conducted by examining in vitro CDK4 and HuR expression through western blotting and qPCR. VPS9D1-AS1 expression was significantly increased in HCC tissues in the analyzed databases and our independent tissue samples. Elevated VPS9D1-AS1 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 VPS9D1-AS1 impaired the proliferative and colony formation activity of HepG2 cells while promoting their apoptotic death. Consistently, VPS9D1-AS1 silencing suppressed HCC tumor growth in vivo. Mechanistically, VPS9D1-AS1 was able to bind to the HuR protein and thereby influence the stability and expression of the CDK4 mRNA, thus impacting HCC cell proliferation. The VPS9D1-AS1/HuR/CDK4 signaling axis regulates HCC tumor cell oncogenic activity, highlighting this pathway as a promising therapeutic target.

中文翻译：

---

lncRNA VPS9D1-AS1通过调节HuR/CDK4轴促进肝细胞癌细胞周期进程

长链非编码 RNA (lncRNA) 代表了与肝细胞癌 (HCC) 相关的有前景的治疗靶点。lncRNA VPS9D1反义RNA 1 ( VPS9D1-AS1 )调节结肠癌和前列腺癌，但其在HCC中的相关性仍有待阐明。使用来自 NCBI 基因表达综合 (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 HCC 中VPS9D1-AS1 的表达。还通过实时定量 PCR (qPCR) 在 80 对 HCC 肿瘤和癌旁组织以及人类 HCC 细胞系中测量了相对 lncRNA 表达。VPS9D1-AS1敲低是通过在体外用特定的 siRNA 构建体转染这些 HCC 细胞来实现的，这些细胞的增殖通过细胞增殖试验和集落形成试验进行量化，而流式细胞术被用来评估它们的细胞周期进程。通过将稳定表达sh-VPS9D1-AS1或对照 shRNA 构建体的HepG2 细胞皮下植入 BALB/c 裸鼠体内，还评估了VPS9D1-AS1 lncRNA 作为 HCC 肿瘤发生调节剂的作用。通过蛋白质印迹和 qPCR检查体外 CDK4和HuR表达，另外进行了机制分析。在分析的数据库和我们的独立组织样本中，HCC 组织中VPS9D1-AS1 的表达显着增加。高架VPS9D1-AS1表达与更大的肿瘤大小和更晚期的肿瘤、淋巴结、转移 (TNM) 分期有关，并且表达更高水平的这种 lncRNA 的 HCC 患者表现出较差的生存结果。敲低VPS9D1-AS1 会损害 HepG2 细胞的增殖和集落形成活性，同时促进它们的凋亡。一致地，VPS9D1-AS1沉默抑制了体内HCC 肿瘤的生长。从机制上讲，VPS9D1-AS1能够与 HuR 蛋白结合，从而影响CDK4 mRNA的稳定性和表达，从而影响 HCC 细胞增殖。该VPS9D1-AS1/HuR/CDK4 信号轴调节 HCC 肿瘤细胞的致癌活性，突出该途径作为有希望的治疗靶点。