正常人骨髓成纤维样基质细胞对骨髓瘤细胞RPMI8226增殖的影响

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RPMI8226增殖的影响，建立了RPMI8226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，Western blot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，RPMI8226细胞的粘附率为29．4％；生长曲线显示直接接触组RPMI8226细胞生长受抑，而非直接接触组(transwell组)无变化；RPMI8226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RPMI8226的增殖，阻止其细胞周期的运行。

Effects of Normal Human Bone Marrow Fibroblastoid Stromal Cell Line on the Proliferation of Multiple Myeloma Cell Line RPMI8226

To investigate the effects of normal human bone marrow fibroblastoid stromal cell lines HFCL on the proliferation of multiple myeloma cell lines RPMI8226,the co-culture system of RPMI8226 with HFCL was established,the adhesion ratio was determined by MTT colorimetric assay, growth curves were determined by trypan blue exclusion,the mitotic index (MI) of RPMI8226 cell was observed by Wright-Giemsa staining. Flow cytometry and Western blot were used to study the changes of cell cycle and expression of proliferating cell nuclear antigen (PCNA),respectively. The results showed that in co-culture the adhesion ratio of RPMI8226 after 1 hour was 29.4%;the proliferation of RPMI8226 cell line in direct contact with HFCL cell line was inhibited as compared with RPMI8226 in suspension. No obvious changes were observed in RPMI8226 cell separated by transwell inserts. The percentage of G_1 phase cells of RPMI8226 cell line in direct contact with HFCL was higher than that of RPMI8226 in suspension,and the percentage of S phase cells was lower. Moreover,the MI of RPMI8226 cell line in suspension was higher than that of RPMI8226 cell line in direct contact with HFCL cell. The expression of PCNA in RPMI8226 cell line in suspension was higher than that of RPMI8226 cell in direct contact with HFCL cell. It is concluded that the normal human bone marrow fibroblastoid stromal cell HFCL inhibits the proliferation and progression of cell cycle of multiple myeloma cell line,RPMI8226.