Abstract：
Hsa_circ_0071589 can exacerbate the malignant behavior of colorectal cancer (CRC) cells. However, its function in stemness and oxaliplatin (OXP) resistance of CRC cells remains unclear. To assess the function of hsa_circ_0071589 in stemness and OXP resistance of CRC cells. Western blotting and qRT-PCR were applied to assess protein and mRNA levels. The association between hsa_circ_0071589, miR-133b and SOX13 was explored via a correlation analysis. Sphere formation was used to assess cell stemness. Meanwhile, the viability of CRC cells and OXP-resistant CRC cells was evaluated with the MTT assay. Cell stemness marker (CD133) levels and apoptosis of CRC cells and OXP-resistant CRC cells were tested using flow cytometry. The ALDH level was investigated using the related detection kit. In addition, the association between hsa_circ_0071589 and miR-133b and SOX13 was investigated using the RIP and dual luciferase assay. Finally, in vivo experiments were performed to detect the function of hsa_circ_0071589 in CRC, and the levels of SOX13, Ki67, and CD44 in mice were evaluated via immunohistochemistry staining. The expression of hsa_circ_0071589 and SOX13 was upregulated in CRC, whereas the expression of miR-133b was downregulated. Hsa_circ_0071589 knockdown significantly inhibited CRC stemness via the mediation of miR-133b. Moreover, hsa_circ_0071589 silencing significantly sensitized CRC cells to OXP by upregulating miR-133b. SOX13 was the direct target of miR-133b, and miR-133b could attenuate stemness and OXP resistance in CRC cells by targeting SOX13. Notably, hsa_circ_0071589 knockdown inhibited tumor growth and decreased OXP resistance in mice with CRC. Hsa_circ_0071589 aggravates stemness and OXP resistance by sponging miR-133b to indirectly target SOX13 in CRC. Thus, our study might present a novel treatment strategy against OXP-resistant CRC.
摘要：
Hsa_circ_0071589可加重结直肠癌(CRC)细胞的恶性行为。然而,其在CRC细胞的干性和奥沙利铂（OXP）耐药性中的功能尚不清楚。评估hsa_circ_0071589在CRC细胞干性和OXP抗性中的功能。应用蛋白质印迹和qRT-PCR评估蛋白质和mRNA水平。通过相关性分析探索hsa_circ_0071589、miR-133b和SOX13之间的关联。球形成用于评估细胞干细胞性。同时,用MTT法评价CRC细胞和OXP耐药CRC细胞的活力.使用流式细胞术测试CRC细胞和OXP抗性CRC细胞的细胞干性标志物(CD133)水平和凋亡。使用相关检测试剂盒调查ALDH水平。此外,使用RIP和双荧光素酶测定研究了hsa_circ_0071589与miR-133b和SOX13之间的关联。最后,进行体内实验以检测hsa_circ_0071589在CRC中的功能，通过免疫组织化学染色评估小鼠体内SOX13，Ki67和CD44的水平。hsa_circ_0071589和SOX13在CRC中的表达上调,而miR-133b的表达下调。Hsa_circ_0071589敲低通过miR-133b的介导显著抑制CRC的干性。此外，hsa_circ_0071589沉默通过上调miR-133b显著地使CRC细胞对OXP敏感。SOX13是miR-133b的直接靶标,miR-133b可通过靶向SOX13减弱CRC细胞的干性和OXP抗性。值得注意的是,hsa_circ_0071589敲除抑制CRC小鼠的肿瘤生长并降低OXP抗性。Hsa_circ_0071589通过使miR-133b间接靶向CRC中的SOX13来加重干性和OXP抗性。因此，我们的研究可能为OXP耐药的CRC提供了一种新的治疗策略.
