Department of Breast and Thyroid Surgery, Liuzhou People's Hospital, NO.8, Wenchang Road, Liuzhou, 545006, Guangxi, People's Republic of China.
Department of Dermatology & Venerology, Affiliated Hospital of Guilin Medical University, Guilin, 541001, Guangxi, People's Republic of China.
Mol Cell Biochem. 2024 Aug;479(8):2055-2068. doi: 10.1007/s11010-023-04819-8. Epub 2023 Jul 31.
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.
hsa_circ_0071589 可加重结直肠癌（CRC）细胞的恶性行为。然而，其在 CRC 细胞干性和奥沙利铂（OXP）耐药中的作用尚不清楚。为了评估 hsa_circ_0071589 在 CRC 细胞干性和 OXP 耐药中的功能。应用 Western blot 和 qRT-PCR 评估蛋白和 mRNA 水平。通过相关性分析探讨 hsa_circ_0071589 与 miR-133b 和 SOX13 的关系。球体形成用于评估细胞干性。同时，MTT 法评估 CRC 细胞和 OXP 耐药 CRC 细胞的活力。用流式细胞术检测 CRC 细胞和 OXP 耐药 CRC 细胞的干细胞标志物（CD133）水平和细胞凋亡。用相关检测试剂盒检测 ALDH 水平。此外，应用 RIP 和双荧光素酶报告基因实验检测 hsa_circ_0071589 与 miR-133b 和 SOX13 的关系。最后，进行体内实验检测 hsa_circ_0071589 在 CRC 中的功能，并通过免疫组化染色评估小鼠中 SOX13、Ki67 和 CD44 的水平。CRC 中 hsa_circ_0071589 和 SOX13 的表达上调，而 miR-133b 的表达下调。hsa_circ_0071589 敲低通过 miR-133b 介导显著抑制 CRC 干性。此外，hsa_circ_0071589 沉默通过上调 miR-133b 显著增加 CRC 细胞对 OXP 的敏感性。SOX13 是 miR-133b 的直接靶标，miR-133b 可通过靶向 SOX13 减轻 CRC 细胞的干性和 OXP 耐药性。值得注意的是，hsa_circ_0071589 敲低抑制了 CRC 小鼠的肿瘤生长并降低了其对 OXP 的耐药性。hsa_circ_0071589 通过海绵吸附 miR-133b 间接靶向 SOX13 加重 CRC 中的干性和 OXP 耐药性。因此，我们的研究可能为 OXP 耐药性 CRC 提供一种新的治疗策略。
