Department of Cardiothoracic Surgery, First Affiliated Hospital of Chengdu Medical College, No. 278, Middle Section of Baoguang Avenue, Xindu District, Chengdu, 610500, Sichuan, China.
Biochem Genet. 2023 Aug;61(4):1351-1368. doi: 10.1007/s10528-022-10307-3. Epub 2022 Dec 31.
Long noncoding RNAs (lncRNAs) are known to participate in the progression of several cancers, including esophageal carcinoma (EC), a common malignancy of the digestive system. Although the role of the lncRNA-miRNA-mRNA regulatory network is crucial for the growth and progression of EC, the regulation of lncRNA BBOX1-AS1 (BBOX1 antisense RNA1) remains unclear. We performed reverse transcription-quantitative PCR (RT-qPCR) and western blotting to evaluate miR-361-3p, collagen type V alpha 1 chain (COL5A1), and BBOX1-AS1 expression levels in EC cells and tissues. The colony formation assay (CFA) and Cell Counting Kit-8 (CCK-8) were employed to identify EC cell proliferation, while western blotting was used to examine EC cell apoptosis and Bax and Bcl-2 expression levels. The effect of BBOX1-AS1 on EC proliferation was determined using an in vivo carcinogenesis assay. Correlation between COL5A1, BBOX1-AS1, and miR-361-3p was examined using the luciferase reporter system and RNA immunoprecipitation assay (RIP). Herein, we observed that BBOX1-AS1 expression levels were upregulated in EC cells and tissues. BBOX1-AS1 knockdown inhibited EC cell proliferation and conferred a pro-apoptotic effect. These results indicated a positive interaction between BBOX1-AS1 and miR-361-3p in EC and a negative association with miR-361-3p. COL5A1 was recognized as a downstream miR-361-3p target and was inversely related to miR-361-3p in EC. Therefore, BBOX1-AS1 expression suppressed cell apoptosis and promoted cell proliferation via the downregulation of miR-361-3p and upregulation of COL5A1 expression. Overall, BBOX1-AS1 facilitates EC progression via the miR-361-3p or COL5A1 axis, indicating that BBOX1-AS1 might be a potential therapeutic target for EC therapy.
长链非编码 RNA（lncRNA）已知参与多种癌症的进展，包括食管癌（EC），这是一种常见的消化系统恶性肿瘤。尽管 lncRNA-miRNA-mRNA 调控网络在 EC 的生长和进展中起着至关重要的作用，但 BBOX1-AS1（BBOX1 反义 RNA1）的调控仍不清楚。我们通过逆转录定量 PCR（RT-qPCR）和 Western 印迹法评估了 EC 细胞和组织中 miR-361-3p、胶原 V 型α1 链（COL5A1）和 BBOX1-AS1 的表达水平。集落形成实验（CFA）和细胞计数试剂盒-8（CCK-8）用于鉴定 EC 细胞增殖，而 Western 印迹法用于检测 EC 细胞凋亡和 Bax 和 Bcl-2 的表达水平。通过体内致癌形成实验确定 BBOX1-AS1 对 EC 增殖的影响。使用荧光素酶报告系统和 RNA 免疫沉淀实验（RIP）检测 COL5A1、BBOX1-AS1 和 miR-361-3p 之间的相关性。在此，我们观察到 BBOX1-AS1 在 EC 细胞和组织中的表达水平上调。BBOX1-AS1 敲低抑制 EC 细胞增殖并发挥促凋亡作用。这些结果表明 BBOX1-AS1 和 miR-361-3p 在 EC 中存在正相互作用，与 miR-361-3p 呈负相关。COL5A1 被认为是 miR-361-3p 的下游靶标，在 EC 中与 miR-361-3p 呈负相关。因此，BBOX1-AS1 通过下调 miR-361-3p 和上调 COL5A1 表达抑制细胞凋亡并促进细胞增殖。总的来说，BBOX1-AS1 通过 miR-361-3p 或 COL5A1 轴促进 EC 进展，表明 BBOX1-AS1 可能是 EC 治疗的潜在治疗靶点。
