Hedgehog信号通路抑制剂HPI-4对软骨肉瘤细胞增殖和侵袭能力的影响

Effect of Hedgehog signaling pathway inhibitor HPI-4 on proliferation and invasion abilities of human chondrosarcoma cells

目的检测Hedgehog(HH)信号通路在人软骨肉瘤中的表达情况,探讨HH信号通路抑制剂HPI-4对于软骨肉瘤细胞SW1353增殖和侵袭能力的影响。方法应用免疫组化技术检测人软骨肉瘤标本中HH信号通路相关蛋白Ihh及增殖相关蛋白Ki67的表达情况,应用CCK-8实验、平板集落形成实验、Transwell肿瘤细胞侵袭实验及免疫印迹法(Western blot)检测HPI-4阻断HH信号通路后SW1353细胞增殖和侵袭能力的变化。结果免疫组化结果证实Ihh蛋白及Ki67蛋白在人软骨肉瘤中存在明显表达。经不同浓度HPI-4干预后,CCK-8实验显示软骨肉瘤细胞SW1353的增殖活性随HPI-4浓度的增高而逐渐降低;平板集落形成实验显示单个肿瘤细胞不断克隆形成集落的能力受到抑制;Transwell实验证实肿瘤细胞侵袭能力随着HPI-4浓度的增高而递减;Western blot结果显示肿瘤细胞内增殖与侵袭相关蛋白的表达明显下降。结论人软骨肉瘤中存在HH信号通路的明显激活,HH信号通路抑制剂HPI-4能够明显抑制软骨肉瘤细胞的增殖和侵袭能力。

Objective To detect the abnormal expression of hedgehog （HH） signaling pathway in human chondrosarcoma cells and discuss the effects of HPI-4 on proliferation and invasion of SW1353 cells.Methods Immunohistochemistry was used to detect the expression of HH related protein Ihh and Ki67.The CCK-8,Tablet colony formation assay,Transwell invasion and Western blot assays were used to detect the effects of HPI-4 on proliferation and invasion abilities of SW1353 cells.Results Immunohistochemical assay confirmed human chondrosarcoma tissue expressed HH signaling related protein Ihh and proliferation related Ki67 obviously.Cell activity experiment showed that after the intervention with HPI-4,proliferation activity of SW1353 cells was gradually decreased with increased drug concentration.At the same time,the plate colony experiments showed a single tumor cell＇s ability of colony was restrained.Transwell experiment confirmed the tumor cell＇s invasion ability was decreased with the increased concentration of HPI-4.Western blotting results showed that HPI-4 could obviously inhibit the expression of tumor cells proliferation and invasion related proteins.Conclusion There was a strong expression of HH signaling pathway in human chondrosarcoma cells and inhibitor HPI-4 could prominently decrease the proliferation and invasion abilities of chondrosarcoma cells.