HIF-1α抑制剂YC-1对人骨肉瘤细胞Saos2和U2-OS增殖和凋亡的影响  

Effects of HIF-1αinhibitor YC-1 on proliferation and apoptosis of human osteosarcoma cell lines Saos2 and U2-OS

作　　者：刘云燕 张瑞 赵景玮 任星光 文艳华 马琼 LIU Yunyan;ZHANG Rui;ZHAO Jingwei;REN Xingguang;WEN Yanhua;MA Qiong(Orthopedic Oncology Institute,the Second Affiliated Hospital of the Air Force Military Medical University,Shaanxi Xi'an 710038,China)

机构地区：[1]空军军医大学第二附属医院骨科实验室,陕西西安710038

出　　处：《现代肿瘤医学》2021年第9期1482-1486,共5页Journal of Modern Oncology

基　　金：国家自然科学基金项目(编号:31670940)。

摘　　要：目的:应用缺氧诱导因子-1α(hypoxia inducible factor-1α,HIF-1α)抑制剂YC-1处理人骨肉瘤细胞Saos2和U2-OS,观察不同药物浓度对人骨肉瘤细胞系Saos2和U2-OS细胞增殖、细胞周期以及凋亡的影响及其机制。方法:人骨肉瘤细胞系Saos2和U2-OS分对照组和实验组。对照组细胞1%氧气浓度低氧培养,实验组细胞施加HIF-1α抑制剂YC-1干预,设浓度梯度。CCK-8法检测不同药物浓度对Saos2和U2-OS细胞活力及增殖的影响;流式细胞术检测YC-1对Saos2和U2-OS细胞周期和凋亡的影响;Western blot对FXR1、G3BP1、APAF1、Cleaved Caspase-3蛋白的表达进行相对定量分析。结果:CCK-8检测结果显示,YC-1对人骨肉瘤Saos2和U2-OS细胞有显著的增殖抑制作用,且呈明显的剂量依赖效应。不同浓度的YC-1处理细胞后,均可使各处理组G1期细胞增多,S期细胞减少,且呈剂量依赖效应(P<0.05)。不同浓度的YC-1处理细胞后,与对照组相比,各实验组FXR1、G3BP1、APAF1蛋白表达显著下降(P<0.05),而Cleaved Caspase-3蛋白表达显著增高(P<0.05)。结论:YC-1可以将人骨肉瘤细胞株Saos2和U2-OS细胞阻滞于G1期,影响细胞周期进程,抑制细胞增殖,促进细胞凋亡。HIF-1α对人骨肉瘤细胞增殖与凋亡的影响可能是通过上调FXR1、G3BP1、APAF1的表达而实现,而YC-1抑制了HIF-1α的表达从而使Cleaved Caspase-3的表达增高诱导细胞凋亡。HIF-1α及FXR1、G3BP1、APAF1、Cleaved Caspase-3有望成为骨肉瘤治疗的新靶标。Objective:To explore the effects and mechanism of YC-1 on the cell proliferation and the apoptosis of human osteosarcoma cell lines Saos2 and U2-OS.Methods:Saos2 and U2-OS cells were divided as control groups and experiment groups.The former were cultured under hypoxia with 1%oxygen,the later were treated by YC-1 with various concentration.Changes of cell cycle and apoptosis rate after treated by YC-1 with various concentration were detected.The effect of YC-1 on the viability and proliferation of human osteosarcoma Saos2 and U2-OS cells were detected by CCK-8 method.The effect of YC-1 on the cycle and apoptosis of human osteosarcoma Saos2 and U2-OS cells were detected by flow cytometry.The FXR1,G3BP1,APAF1,Cleaved Caspase-3 protein was evaluated by Western blot.Results:CCK-8 assay showed that YC-1 significantly inhibited the proliferation of human osteosarcoma Saos2 and U2-OS cells in concentration dependent manner.The distribution of cell cycle by flow cytometry indicated that after treated with YC-1,the number of cells in G1 phase increased gradually,while the number of cells in S phase decreased,which was in a dose dependent manner.Following the increasing of the concentration of YC-1,compared with control group,expression of FXR1,G3BP1,APAF1 was decreased significantly(P<0.05),but Cleaved Caspase-3 protein increased significantly(P<0.05).Conclusion:YC-1 could inhibit the proliferation and affect the cell cycle by inhibiting the G1 period of Saos2 and U2-OS cells.The mechanisms are supposed to be that HIF-1αup-regulates the expression of FXR1,G3BP1,APAF1.YC-1 inhibites the expression of HIF-1α,increases the expression of Cleaved Caspase-3 and induces apoptosis.The application of the HIF-1αin hibitor offers a new way for the treatment of the osteosarcoma.

关 键 词：人骨肉瘤细胞 缺氧诱导因子抑制剂 增殖 凋亡 

分 类 号：R738.1[医药卫生—肿瘤]