Department of General Medicine, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430000, Hubei, China.
Curr Mol Pharmacol. 2023;16(8):894-904. doi: 10.2174/1874467216666230130132927.
This study aimed to unveil the specific function of lncRNA BBOX1 antisense RNA 1 (BBOX1-AS1) in ESCC cells and the underlying regulatory mechanism.
Esophageal squamous cell carcinoma (ESCC) is a deadly disease. Molecular mechanisms essential to ESCC development and progression require in-depth investigation. Long noncoding RNAs (lncRNAs) have been suggested as crucial effectors in modulating tumor growth.
RT-qPCR and western blot examined the expression of genes and proteins of concern, respectively. Colony formation and EdU assays assessed the changes in cell proliferation. Sphere formation assay also detected the stemness of ESCC cells. Bioinformatics prediction, along with mechanistic assays (FISH, Subcellular fractionation, RNA pull-down, RIP, and luciferase reporter), was conducted to explore the gene interactions and regulatory relationship.
BBOX1-AS1 was observed to be aberrantly up-regulated in ESCC tissues and cell lines. BBOX1-AS1 depletion exerted suppressive impacts on ESCC cell proliferation and stemness, while BBOX1-AS1 overexpression led to the opposite consequences. Moreover, BBOX1-AS1 was verified to activate Hedgehog signaling pathway via up-regulating PTCH1, and BBOX1-AS1 could sponge miR-506-5p to up-regulate EIF5A, thus stabilizing PTCH1 mRNA. Rescue experiments indicated that BBOX1-AS1 could affect ESCC cell proliferation and stemness via modulation on PTCH1.
To conclude, BBOX1-AS1 activates Hedgehog signaling pathway to facilitate the proliferation and stemness of ESCC cells via miR-506-5p/EIF5A/PTCH1 axis.
本研究旨在揭示长链非编码 RNA（lncRNA）BBOX1 反义 RNA 1（BBOX1-AS1）在 ESCC 细胞中的特定功能及其潜在的调控机制。
食管鳞状细胞癌（ESCC）是一种致命的疾病。ESCC 发展和进展所必需的分子机制需要深入研究。长非编码 RNA（lncRNAs）已被认为是调节肿瘤生长的关键效应物。
分别采用 RT-qPCR 和 Western blot 检测相关基因和蛋白的表达。集落形成和 EdU 检测评估细胞增殖的变化。球形成实验也检测了 ESCC 细胞的干性。通过生物信息学预测以及机制研究（FISH、亚细胞分离、RNA 下拉、RIP 和荧光素酶报告基因），探索基因相互作用和调控关系。
BBOX1-AS1 在 ESCC 组织和细胞系中异常上调。BBOX1-AS1 的耗竭对 ESCC 细胞增殖和干性具有抑制作用，而 BBOX1-AS1 的过表达则导致相反的结果。此外，BBOX1-AS1 通过上调 PTCH1 激活 Hedgehog 信号通路，并且 BBOX1-AS1 可以海绵吸附 miR-506-5p 来上调 EIF5A，从而稳定 PTCH1 mRNA。挽救实验表明，BBOX1-AS1 通过调节 PTCH1 影响 ESCC 细胞增殖和干性。
总之，BBOX1-AS1 通过 miR-506-5p/EIF5A/PTCH1 轴激活 Hedgehog 信号通路促进 ESCC 细胞的增殖和干性。
