舒尼替尼调控骨肉瘤细胞的增殖、迁移与凋亡机制研究

On Mechanism of Sunitinib Regulating Cell Proliferation,Migration and Apoptosis in Osteosarcoma

目的探讨抗肿瘤药物舒尼替尼对骨肉瘤细胞增殖、迁移与凋亡的影响及潜在作用机制.方法制作骨肉瘤原位癌和转移癌的小鼠模型,将模型小鼠分为A组(40 mg/kg舒尼替尼)、B组(10 mg/kg PD-L1抗体)、C组(舒尼替尼和PD-L1抗体联合治疗)、D组(PBS对照);定期测量肿瘤体积大小,通过观察转移的骨肉瘤小鼠肺组织,确定骨肉瘤细胞的百分比;通过流式细胞学染色技术观察小鼠的免疫细胞群数量,明确肿瘤的免疫微环境;通过Western blot方法测定骨肉瘤的PD-L1和STAT3,通过CCK-8和Transwell法分析骨肉瘤的侵袭性.结果PD-L1表达水平在各骨肉瘤细胞间比较差异具有统计学意义(P<0.05),其中KRIB、U2OS、143B和LM7细胞中表达水平较高,而在MG63.2和SAOS-2细胞中的表达水平较低;舒尼替尼可显著降低KRIB和LM7细胞的转移和侵袭能力,有效降低骨肉瘤细胞的转移和侵袭;舒尼替尼通过调控STAT3而降低PD-L1的表达,并且还能重建实验小鼠的免疫系统,有效控制骨肉瘤的转移;舒尼替尼和PD-L1抗体联合应用可以更好地控制骨肉瘤生长与远处转移.结论舒尼替尼可有效控制荷瘤小鼠骨肉瘤的增殖与迁移.

Objective To investigate the effects of Sunitinib on proliferation,migration and apoptosis of osteos-arcoma cells and its potential mechanism.Method Mouse models of osteosarcoma carcinoma in situ and metastatic carcinoma were made and divided into group A(Sunitinib 40 mg/kg),group B(10 mg/kg PD-L1 antibody),group C(combination therapy),group D(PBS control).Tumor volume was measured periodically,and the percentage of osteosarcoma cells was determined by observing lung tissue of metastatic osteosarcoma mice.Flow cytometry was used to observe the number of immune cell populations in mice,and the changes of immune microenvironment of tumor were identified.PD-L1 and STAT3 of osteosarcoma were determined by Western blot,and the aggressiveness of osteosarcoma was investigated by CCK-8 and Transwell method.Results There was significant difference in the expression level of PD-L1 among osteosarcoma cells(P<0.05);The expression levels of KRIB,U2OS,143B and LM7 cells were higher,while the expression levels were lower in MG63.2 and SAOS-2 cells.Sunitinib can significantly reduce the metastasis and invasion of KRIB and LM7 cells.Sunitinib can effectively reduce the metastasis and invasion of osteosarcoma cells.Sunitinib can reduce the expression of PD-L1 by regulating the activation of STAT3,and can reconstruct the immune system of tumor-bearing mice,and effectively control the metastasis of osteosarcoma.The combined application of Sunitinib and PD-L1 antibodies can better control the growth and distant metastasis of osteosarcoma.Conclusion Sunitinib can effectively control the proliferation and migration of osteosarcoma in tumor-bearing mice.