Xu Jiuhui, Chen Chenglong, Sun Kunkun, Shi Qianyu, Wang Boyang, Huang Yi, Ren Tingting, Tang Xiaodong
Musculoskeletal Tumor Center, Peking University People's Hospital, Beijing, China.
Beijing Key Laboratory of Musculoskeletal Tumor, Peking University People's Hospital, Beijing, China.
Front Oncol. 2023 Jun 19;13:1192472. doi: 10.3389/fonc.2023.1192472. eCollection 2023.
Anlotinib, a tyrosine kinase inhibitor (TKI) has been in clinical application to inhibit malignant cell growth and lung metastasis in osteosarcoma (OS). However, a variety of drug resistance phenomena have been observed in the treatment. We aim to explore the new target to reverse anlotinib resistance in OS.
In this study, we established four OS anlotinib-resistant cell lines, and RNA-sequence was performed to evaluate differentially expressed genes. We verified the results of RNA-sequence by PCR, western blot and ELISA assay. We further explored the effects of tocilizumab (anti- IL-6 receptor), either alone or in combined with anlotinib, on the inhibition of anlotinib-resistant OS cells malignant viability by CCK8, EDU, colony formation, apoptosis, transwell, wound healing, Cytoskeletal stain assays, and xenograft nude mouse model. The expression of IL-6 in 104 osteosarcoma samples was tested by IHC.
We found IL-6 and its downstream pathway STAT3 were activated in anlotinib-resistant osteosarcoma. Tocilizumab impaired the tumor progression of anlotinib-resistant OS cells, and combined treatment with anlotinib augmented these effects by inhibiting STAT3 expressions. IL-6 was highly expressed in patients with OS and correlated with poor prognosis.
Tocilizumab could reverse anlotinib resistance in OS by IL-6/STAT3 pathway and the combination treatment with anlotinib rationalized further studies and clinical treatment of OS.
安罗替尼是一种酪氨酸激酶抑制剂（TKI），已在临床应用中用于抑制骨肉瘤（OS）中的恶性细胞生长和肺转移。然而，在治疗过程中观察到了多种耐药现象。我们旨在探索逆转OS中安罗替尼耐药的新靶点。
在本研究中，我们建立了四种OS安罗替尼耐药细胞系，并进行RNA测序以评估差异表达基因。我们通过PCR、蛋白质免疫印迹和酶联免疫吸附测定法验证了RNA测序结果。我们进一步通过CCK8、EDU、集落形成、凋亡、Transwell、伤口愈合、细胞骨架染色试验以及异种移植裸鼠模型，探讨了托珠单抗（抗IL-6受体）单独或与安罗替尼联合使用对抑制安罗替尼耐药OS细胞恶性活力的影响。通过免疫组织化学检测了104例骨肉瘤样本中IL-6的表达。
我们发现IL-6及其下游通路STAT3在安罗替尼耐药的骨肉瘤中被激活。托珠单抗损害了安罗替尼耐药OS细胞的肿瘤进展，与安罗替尼联合治疗通过抑制STAT3表达增强了这些作用。IL-6在OS患者中高表达，且与预后不良相关。
托珠单抗可通过IL-6/STAT3通路逆转OS中的安罗替尼耐药，与安罗替尼联合治疗为OS的进一步研究和临床治疗提供了理论依据。
