中华血液学杂志
中華血液學雜誌
중화혈액학잡지
Chinese Journal of Hematology
2010年
10期
680-683
,共4页
郝牧%谢振卿%韩有金%安刚%孟恒星%黄靖%李长虹%邹德慧%邱录贵
郝牧%謝振卿%韓有金%安剛%孟恆星%黃靖%李長虹%鄒德慧%邱錄貴
학목%사진경%한유금%안강%맹항성%황정%리장홍%추덕혜%구록귀
骨髓微环境%多发性骨髓瘤%间充质干细胞%细胞凋亡
骨髓微環境%多髮性骨髓瘤%間充質榦細胞%細胞凋亡
골수미배경%다발성골수류%간충질간세포%세포조망
Bone marrow microenvironment%Multiple myeloma%Mesenchymal stem cell%Cell apoptosis
目的 探讨间充质干细胞在多发性骨髓瘤细胞生长以及硼替佐米诱导骨髓瘤细胞凋亡中的作用.方法 取15例临床确诊的多发性骨髓瘤(MM)患者以及3名正常供者的骨髓标本,分离其间充质干细胞(MM-BMSC和ND-BMSC);测定细胞生长曲线、免疫表型和细胞因子分泌水平.将骨髓瘤细胞(NCI-H929)与BMSC细胞共培养,并加入蛋白酶体抑制剂硼替佐米,观察BMSC对NCI-H929细胞生长增殖的影响;进一步通过Annexin V-FITC/PI双染法流式细胞术(FCM)检测BMSC对硼替佐米诱导NCI-H929细胞凋亡的影响.结果 成功分离得到MM患者以及正常供者骨髓间充质干细胞,FCM检测显示两者均高表达CD73和CD105(＞95%),表达CD44和CD29,不表达CD31、CD34、CD45和HLA-DR(＜1%)等表面分子.生长曲线测定显示MM-BMSC增殖较为缓慢,倍增时间(82 h)较ND-BMSC(62 h)略有延长(P＜0.05).与ND-BMSC比较,MM-BMSC分泌高水平的细胞因子IL-6和VEGF,分别为(188.8±9.4)pg/ml对(115.0±15.1)pg/ml和(1497.2±39.7)pg/ml对(1329.0±21.1)pg/ml.将BMSC与骨髓瘤细胞NCI-H929共培养,MM-BMSC可促进骨髓瘤细胞的生存,降低NCI-H929细胞对硼替佐米的敏感性,明显抑制硼替佐米诱导的瘤细胞凋亡.结论 MM-BMSC可促进骨髓瘤细胞的生长,明显减少蛋白酶体抑制剂硼替佐米诱导的细胞凋亡.
目的 探討間充質榦細胞在多髮性骨髓瘤細胞生長以及硼替佐米誘導骨髓瘤細胞凋亡中的作用.方法 取15例臨床確診的多髮性骨髓瘤(MM)患者以及3名正常供者的骨髓標本,分離其間充質榦細胞(MM-BMSC和ND-BMSC);測定細胞生長麯線、免疫錶型和細胞因子分泌水平.將骨髓瘤細胞(NCI-H929)與BMSC細胞共培養,併加入蛋白酶體抑製劑硼替佐米,觀察BMSC對NCI-H929細胞生長增殖的影響;進一步通過Annexin V-FITC/PI雙染法流式細胞術(FCM)檢測BMSC對硼替佐米誘導NCI-H929細胞凋亡的影響.結果 成功分離得到MM患者以及正常供者骨髓間充質榦細胞,FCM檢測顯示兩者均高錶達CD73和CD105(＞95%),錶達CD44和CD29,不錶達CD31、CD34、CD45和HLA-DR(＜1%)等錶麵分子.生長麯線測定顯示MM-BMSC增殖較為緩慢,倍增時間(82 h)較ND-BMSC(62 h)略有延長(P＜0.05).與ND-BMSC比較,MM-BMSC分泌高水平的細胞因子IL-6和VEGF,分彆為(188.8±9.4)pg/ml對(115.0±15.1)pg/ml和(1497.2±39.7)pg/ml對(1329.0±21.1)pg/ml.將BMSC與骨髓瘤細胞NCI-H929共培養,MM-BMSC可促進骨髓瘤細胞的生存,降低NCI-H929細胞對硼替佐米的敏感性,明顯抑製硼替佐米誘導的瘤細胞凋亡.結論 MM-BMSC可促進骨髓瘤細胞的生長,明顯減少蛋白酶體抑製劑硼替佐米誘導的細胞凋亡.
목적 탐토간충질간세포재다발성골수류세포생장이급붕체좌미유도골수류세포조망중적작용.방법 취15례림상학진적다발성골수류(MM)환자이급3명정상공자적골수표본,분리기간충질간세포(MM-BMSC화ND-BMSC);측정세포생장곡선、면역표형화세포인자분비수평.장골수류세포(NCI-H929)여BMSC세포공배양,병가입단백매체억제제붕체좌미,관찰BMSC대NCI-H929세포생장증식적영향;진일보통과Annexin V-FITC/PI쌍염법류식세포술(FCM)검측BMSC대붕체좌미유도NCI-H929세포조망적영향.결과 성공분리득도MM환자이급정상공자골수간충질간세포,FCM검측현시량자균고표체CD73화CD105(＞95%),표체CD44화CD29,불표체CD31、CD34、CD45화HLA-DR(＜1%)등표면분자.생장곡선측정현시MM-BMSC증식교위완만,배증시간(82 h)교ND-BMSC(62 h)략유연장(P＜0.05).여ND-BMSC비교,MM-BMSC분비고수평적세포인자IL-6화VEGF,분별위(188.8±9.4)pg/ml대(115.0±15.1)pg/ml화(1497.2±39.7)pg/ml대(1329.0±21.1)pg/ml.장BMSC여골수류세포NCI-H929공배양,MM-BMSC가촉진골수류세포적생존,강저NCI-H929세포대붕체좌미적민감성,명현억제붕체좌미유도적류세포조망.결론 MM-BMSC가촉진골수류세포적생장,명현감소단백매체억제제붕체좌미유도적세포조망.
Objective To investigate the role of mesenchymal stem cells(BMSCs)in multiple myeloma(MM)bone marrow(BM)microenrivonment and their effect on myeloma cells survival and bortezomib induced apoptosis.Methods BMSCs were derived from BM of untreated myeloma patients(MM-BMSCs)and healthy donors(HD-BMSCs),respectively.The phenotype,proliferation time and cytokine secretion of MM-BMSCs were detected and compared with HD-BMSCs.Then BMSCs were co-cultured with myeloma cell line NCI-H929 and bortezomib in vitro.The NCI-H929 cells proliferation and bortezomib induced cell apoptosis were investigated.Results MM-BMSCs and HD-BMSCs were isolated successfully.The phenotype of MM-BMSCs was similar to that of HD-BMSCs.Expressions of CD73,CD105,CD44 and CD29 were positive,but those of CD31,CD34,CD45 and HLA-DR(＜ 1%)negative.The proliferation time of MM-BMSCs was longer than that of HD-BMSCs(82 h vs 62 h,P ＜ 0.05).Moreover,over-expressions of IL-6 and VEGF in MM-BMSCs culture supernatant were detected as compared with that in HD-BMSCs [(188.8 ±9.4)pg/ml vs(115.0±15.1)pg/ml and(1497.2 ±39.7)pg/ml vs(1329.0±21.1)pg/ml,respectively].MM-BMSCs supported survival of the myeloma cells NCI-H929 and protected them from bortezomib induced cell apoptosis.Conclusions MM-BMSCs is benefit for myeloma cells proliferation and against cell apoptosis induced by bortezomib.Over-expression of IL-6 and VEGF maybe play a critical role in these effects.
