Zhao Fuyou, Wu Qiong, Dai Xiusong, Li Yumei, Gan Huaiyong, Wang Ri, Lv Jie, Chen Yuqing
Department of Medical Oncology, The First Affiliated Hospital of Bengbu Medical College, Bengbu, Anhui 233000, P.R. China.
Department of Orthopedics, The First Affiliated Hospital of Bengbu Medical College, Bengbu, Anhui 233000, P.R. China.
Oncol Lett. 2016 Dec;12(6):5199-5204. doi: 10.3892/ol.2016.5311. Epub 2016 Oct 24.
The aim of the present study was to investigate the effect of programmed cell death 1 (PD-1) on osteosarcoma (OD) stem cells and T cells, and to determine their correlation. OS stem cells were sorted and identified from OS MG63 cells. Flow cytometry was used to detect the PD-1 expression of the OS tumor stem cell membrane surface. The expression of PD-1 mRNA was detected by reverse transcription-polymerase chain reaction (RT-PCR). MTT was used to detect the effect of PD-1 signals on T-cell proliferation. The results indicated that the cancer cells (cultured in DMEM medium containing 10% fetal bovine serum) exhibited clear proliferation within 1 week of cell culture, which showed their strong proliferation and aggressive ability. The formation of tumor cell spheres was dependent on the support of serum nutrition. The proliferation of MG63 cells in the serum culture medium was significantly higher than the number of OS cell spheres in serum-free suspension culture (P<0.05). Pluripotent stem cells in cancer cell spheres exhibited significantly higher cluster of differentiation 133 expression compared with the MG63 cells. The PD-1 expression levels of the cancer cell spheres was significantly increased compared with the MG63 cells, which is consistent with the results of the RT-PCR. In conclusion, the MG63 cell line possesses the features of OS stem cells. The MG63 cell line can express the certain cancer-associated cell markers. The expression of PD-1 in spheres was also increased significantly compared to the MG63 cells, which can reduce the immune function of patients and may be closely associated with the occurrence and development of tumors.
本研究的目的是探讨程序性细胞死亡蛋白1(PD-1)对骨肉瘤(OS)干细胞和T细胞的影响,并确定它们之间的相关性。从骨肉瘤MG63细胞中筛选并鉴定出OS干细胞。采用流式细胞术检测OS肿瘤干细胞膜表面PD-1的表达。通过逆转录-聚合酶链反应(RT-PCR)检测PD-1 mRNA的表达。采用MTT法检测PD-1信号对T细胞增殖的影响。结果表明,癌细胞(在含10%胎牛血清的DMEM培养基中培养)在细胞培养1周内表现出明显的增殖,显示出其较强的增殖和侵袭能力。肿瘤细胞球的形成依赖于血清营养的支持。血清培养基中MG63细胞的增殖明显高于无血清悬浮培养中OS细胞球的数量(P<0.05)。与MG63细胞相比,癌细胞球中的多能干细胞表现出明显更高的分化簇133表达。与MG63细胞相比,癌细胞球的PD-1表达水平显著升高,这与RT-PCR结果一致。综上所述,MG63细胞系具有OS干细胞的特征。MG63细胞系可表达某些癌症相关细胞标志物。与MG63细胞相比,球状体中PD-1的表达也显著增加,这可能会降低患者的免疫功能,并且可能与肿瘤的发生发展密切相关。