PD-1信号对骨肉瘤类肿瘤干细胞及T细胞增殖的影响

Effect of PD-1 signal on the proliferation of human osteosarcoma cells and T cells

目的探讨程序性死亡-1(PD-1)对骨肉瘤细胞干细胞及T细胞增殖的影响。方法对人骨肉瘤细胞株MG-63细胞进行分选及球体诱导,采用MTT法检测骨肉瘤MG-63与肿瘤细胞球的增殖过程;采用实时荧光定量聚合酶链反应检测PD-1 mRNA的表达。结果7 d内癌细胞明显增殖,显示较强的增殖能力和侵袭性;肿瘤细胞球的形成依赖于血清营养的支持,与无血清悬浮培养的骨肉瘤细胞球细胞数量相比,血清培养基中的MG-63癌细胞增殖数量显著较高(P<0.05);实时荧光定量聚合酶链反应检测结果显示多能干细胞标记CD133在癌细胞球中的表达显著高于MG-63,癌细胞球及MG-63中PD-1表达均明显增加(P<0.05)。结论MG-63细胞系能够表达相应的细胞标记,随着骨肉瘤细胞的增殖PD-1的表达增加,免疫功能下降,说明PD-1表达与骨肉瘤的进展过程密切相关。

Objective To investigate the influences of Programmed death-1(PD-1)on the proliferation of osteosarcoma cells and T cells proliferation.Methods The human osteosarcoma cell line MG-63 cells were sorting and induced to cell spheres.Qsteosarcoma MG-63 and tumor cell spheres were detected by MTT method.The expression of PD-1 mRNA were detected by RT-PCR.Results After 7 days,the cancer cells had an obvious proliferation,which showed strong ability of proliferation and invasion.The formation of tumor cell spheres depends on the support of serum nutrition;compared with the number of MG-63 cells proliferation in serum-free suspension culture,the number was significantly higher in the serum suspension(P<0.05).RT-PCR results showed that The expression of the pluripotent stem cell marker CD133 in cancer cells was significantly higher than that in MG-63,the PD-1 expression of cancer cells spheres and MG-63 were both increased significantly(P<0.05).Conclusion The MG-63 cell line could express the corresponding cell markers.With the proliferation of osteosarcoma cells the expression of PD-1 increased,the immune function decreased.All of above showed that the expression PD-1 closely related to the progression of osteosarcoma.