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为了观察正常人骨髓成纤维样基质细胞系HFCL对多发性骨髓瘤细胞系RPMI82 2 6增殖的影响 ,建立了RPMI82 2 6细胞和HFCL细胞共培养体系 ,用MTT法测粘附率 ,台盼蓝拒染法测定生长曲线 ;流式细胞术 (FCM )检测细胞周期的变化 ,瑞氏 吉姆萨染色后进行分裂指数 (MI)测定 ,Westernblot检测增殖细胞核抗原 (PCNA)表达。结果显示 ,与HFCL细胞共培养 1小时后 ,RPMI82 2 6细胞的粘附率为 2 9.4 % ;生长曲线显示直接接触组RPMI82 2 6细胞生长受抑 ,而非直接接触组 (transwell组 )无变化 ;RPMI82 2 6细胞与HFCL细胞共培养后 ,直接接触组G1期细胞增高 ,S期细胞减少 ;其分裂指数表现为单独骨髓瘤细胞组大于与HFCL直接接触组 ;直接接触组PCNA表达下调。结论 :正常人骨髓成纤维样基质细胞HFCL能抑制骨髓瘤细胞系RPMI82 2 6的增殖 ,阻止其细胞周期的运行。
In order to observe the effect of normal human bone marrow fibroblastoid stromal cell line HFCL on the proliferation of multiple myeloma RPMI 82 26 cells, a co-culture system of RPMI 8226 cells and HFCL cells was established. The adhesion rate, trypan blue Flow cytometry (FCM) was used to detect cell cycle changes. Wright’s Giemsa staining was used to determine the mitotic index (MI), and Western blot was used to detect the expression of proliferating cell nuclear antigen (PCNA). The results showed that the adhesion rate of RPMI8226 cells was 2 9.4% after 1 h co-culture with HFCL cells. The growth curve showed that the growth of RPMI82 2 6 cells was inhibited in the direct contact group, but not in the non-direct contact group (transwell group) After co-cultured with HFCL cells, the number of cells in G1 phase of RPMI82 2 6 cells increased and the number of S phase cells decreased in the direct contact group. The split index showed that the myeloma cell group was more than the direct contact group with HFCL; Conclusion: HFCL in normal human bone marrow fibroblastoid stromal cells can inhibit the proliferation of myeloma cell line RPMI82 2 6 and prevent its cell cycle operation.