An Shixiang, Cui Jiaxian, Yang Wenhong, Zhang Mingyu, Yu Huiling, Lu Jingkun, Tian Yunpeng, Qiao Lu, Wang Xiumei, Bao Lili, Zhao Pengwei
Laboratory of Microbiology and Immunology, School of Basic Medical Science, Inner Mongolia Medical University, Xinhua Street, Hohhot, 010059, PR China.
Medical Oncology, Affiliated People's Hospital of Inner Mongolia Medical University, Hohhot, Inner Mongolia, 010020, China.
J Transl Med. 2025 Mar 6;23(1):281. doi: 10.1186/s12967-024-05860-x.
Among all malignancies, colorectal cancer ranks third in incidence rate and second in mortality rate. Human beta-defensin-1 (hBD-1) has broad-spectrum antimicrobial properties, and it plays an important role in the tumor microenvironment. Circular ribonucleic acids (circRNAs) regulate the proliferation and progression of colorectal cancer cells via cancer-related signaling pathways.
Cell proliferation was assessed using the Cell Counting Kit-8 assay to determine the optimal hBD-1 concentration. Intracellular autophagic vesicles were visualized via monodansylcadaverine staining. In addition, the levels of AKT and mammalian target of rapamycin (mTOR)-associated signaling proteins were analyzed via Western blot analysis. CircRNA microarrays and quantitative real-time polymerase chain reaction were used to identify differentially expressed circRNAs in colon cancer cell lines. The functional role of HAS-CIRCpedia-5280 in vitro was demonstrated by overexpressing HAS-CIRCpedia-5280 and inhibiting miR-4712-5p. HAS-CIRCpedia-5280 could be a sponge of miR-4712-5p, mimicking the effect induced by HAS-CIRCpedia-5280 overexpression in colon cancer cells.
hBD-1 inhibited the proliferation of colon cancer cells and increased the number of intracellular autophagic vesicles. In addition, hBD-1 inhibited the AKT/mTOR signaling pathway, thereby enhancing cellular autophagy. Further, the interaction of HAS-CIRCpedia-5280 and miR-4712-5p was investigated. hBD-1 upregulated the expression level of HAS-CIRCpedia-5280 and downregulated the expression level of miR-4712-5p in colon cancer cells. Subsequently, the overexpression of HAS-CIRCpedia-5280 or the inhibition of miR-4712-5p activated the AKT/mTOR signaling pathway, leading to cellular autophagy inhibition. Conversely, the mimicry of miR-4712-p counteracted the effect of HAS-CIRCpedia 5280 overexpression in colon cancer cells by inhibiting the activation of the AKT/mTOR signaling pathway and, thereby, enhancing cellular autophagy.
hBD-1 can have an inhibitory effect against cell proliferation in colon cancer SW-620/HCT-116 cells via the HAS-CIRCpedia-5280/miR-4712-5p-mediated activation of autophagy.
在所有恶性肿瘤中，结直肠癌的发病率排名第三，死亡率排名第二。人β-防御素-1（hBD-1）具有广谱抗菌特性，且在肿瘤微环境中发挥重要作用。环状核糖核酸（circRNAs）通过癌症相关信号通路调节结肠癌细胞的增殖和进展。
使用细胞计数试剂盒-8检测法评估细胞增殖，以确定最佳hBD-1浓度。通过单丹磺酰尸胺染色观察细胞内自噬泡。此外，通过蛋白质免疫印迹分析检测AKT和雷帕霉素哺乳动物靶蛋白（mTOR）相关信号蛋白的水平。使用circRNA微阵列和定量实时聚合酶链反应鉴定结肠癌细胞系中差异表达的circRNAs。通过过表达HAS-CIRCpedia-5280和抑制miR-4712-5p来证明HAS-CIRCpedia-5280在体外的功能作用。HAS-CIRCpedia-5280可能是miR-4712-5p的海绵，模拟HAS-CIRCpedia-5280在结肠癌细胞中过表达所诱导的效应。
hBD-1抑制结肠癌细胞的增殖并增加细胞内自噬泡的数量。此外，hBD-1抑制AKT/mTOR信号通路，从而增强细胞自噬。进一步研究了HAS-CIRCpedia-5280与miR-4712-5p的相互作用。hBD-1上调结肠癌细胞中HAS-CIRCpedia-5280的表达水平并下调miR-4712-5p的表达水平。随后，HAS-CIRCpedia-5280的过表达或miR-4712-5p的抑制激活了AKT/mTOR信号通路，导致细胞自噬受到抑制。相反，miR-4712-5p模拟物通过抑制AKT/mTOR信号通路的激活来抵消HAS-CIRCpedia 5280在结肠癌细胞中过表达的作用，从而增强细胞自噬。
hBD-1可通过HAS-CIRCpedia-5280/miR-4712-5p介导的自噬激活对结肠癌SW-620/HCT-116细胞的增殖产生抑制作用。
