摘要
目的探讨高三尖杉酯碱(HHT)对骨肉瘤细胞的抗肿瘤作用,并探讨其可能的作用靶点和机制。方法使用不同浓度的高三尖杉酯碱处理骨肉瘤细胞系(143b及U2-OS),通过CCK-8实验检测HHT对骨肉瘤细胞的抗肿瘤作用,集落形成实验检测骨肉瘤细胞的克隆能力,划痕实验检测骨肉瘤细胞的迁移能力,Transwell实验检测骨肉瘤细胞的侵袭能力,流式细胞学技术检测骨肉瘤细胞的凋亡率。并通过荧光定量PCR(qPCR)实验检测经过HHT处理后骨肉瘤细胞内基因表达情况的改变,并找出HHT作用于骨肉瘤细胞内可能的靶点和机制。结果CCK-8实验证实HHT对骨肉瘤细胞143b和U2-OS的增殖具有明显的抑制作用;集落形成实验、划痕实验、Transwell实验分别证实了HHT对骨肉瘤细胞143b和U2-OS的克隆能力、迁移能力、侵袭能力存在显著的抑制作用;流式细胞学技术证实了HHT能提高骨肉瘤细胞143b和U2-OS的凋亡率;RT-qPCR实验表明HHT可以显著上调casp3和casp8的表达,验证了HHT可能是通过调控casp3和casp8进而对骨肉瘤的增殖凋亡、侵袭及转移等发挥作用。结论casp3和casp8是HHT作用于骨肉瘤细胞的靶点,HHT通过上调casp3和casp8的表达来发挥对骨肉瘤细胞的抑制作用。
Objective To explore the anti-tumour effect of homoharringtonine(HHT)on osteosarcoma cells and its possible target and mechanism.Methods Osteosarcoma cell lines(143b and U2-OS)were treated with different concentrations of HHT.The anti-tumour effect of HHT on osteosarcoma cells was detected by CCK-8 assay,the cloning ability of osteosarcoma cells was detected by colony formation assay,and the migration ability of osteosarcoma cells was detected by scratching assay.The invasion ability of osteosarcoma cells was detected by Transwell assay,and the apoptosis rate of osteosarcoma cells was detected by flow cytometry.The changes of gene expression in osteosarcoma cells after HHT treatment were detected by fluorescence quantitative PCR(q PCR),and the possible targets and mechanisms of HHT action on osteosarcoma cells were identified.Results CCK-8 assay confirmed that HHT had obvious inhibition of proliferation on osteosarcoma cells 143b and U2-OS.Colony formation assay,scratching assay and Transwell assay confirmed that HHT significantly inhibited the cloning ability,migration ability and invasion ability of osteosarcoma cells 143b and U2-OS.Flow cytometry confirmed that HHT could increase the apoptosis rate of osteosarcoma cells 143b and U2-OS.RT-q PCR showed that HHT could significantly up-regulate the expression of casp3 and casp8,confirming that HHT might play a role in the proliferation,apoptosis,invasion and metastasis of osteosarcoma by regulating casp3 and casp8.Conclusion Casp3and casp8 may be the target of HHT on osteosarcoma cells,and HHT exerts its inhibitory effect on osteosarcoma cells by up-regulating the expression of casp3 and casp8.
作者
钟厚成
朱宇凡
杨敏
魏任雄
Zhong Houcheng;Zhu Yufan;Yang Min;Wei Renxiong(Department of Spinal and Bone Oncology,Zhongnan Hospital of Wuhan University,Wuhan Hubei,430072,China)
出处
《生物骨科材料与临床研究》
CAS
2022年第4期8-15,共8页
Orthopaedic Biomechanics Materials and Clinical Study
