中华血液学杂志
中華血液學雜誌
중화혈액학잡지
Chinese Journal of Hematology
2014年
7期
641-644
,共4页
芮明忠%付天文%刘英%黄海雯%傅晋翔
芮明忠%付天文%劉英%黃海雯%傅晉翔
예명충%부천문%류영%황해문%부진상
罗格列酮%多发性骨髓瘤%基因,HIF1α%基因,IGF1
囉格列酮%多髮性骨髓瘤%基因,HIF1α%基因,IGF1
라격렬동%다발성골수류%기인,HIF1α%기인,IGF1
Rosiglitazone%Multiple myeloma%Gene,HIF1α%Gene,IGF1
目的 观察罗格列酮作用后骨髓瘤细胞HIF1α和IGF1 mRNA表达水平,探讨罗格列酮抑制骨髓瘤血管形成的可能机制.方法 以骨髓瘤细胞系RPMI 8226细胞及5例初诊多发性骨髓瘤患者骨髓浆细胞(CD138免疫磁珠筛选,作为骨髓瘤原代细胞)为研究对象,采用RT-PCR法检测用不同浓度(10、20、40 μmol/L)罗格列酮处理前后骨髓瘤细胞HIF1α、IGF1 mRNA表达水平;采用Westernblot法检测AKT和ERK蛋白磷酸化和非磷酸化表达的情况.同时以5例缺铁性贫血患者的骨髓单个核细胞为对照组.结果 RPMI8226细胞和骨髓瘤原代细胞HIF1α和IGF1 mRNA表达显著高于对照组.罗格列酮浓度为20 μmol/L处理48 h后,与对照组相比,RPMI 8226细胞中HIF1α、IGF1 mRNA相对表达水平分别由1.21±0.08和0.62±0.06降至0.75±0.06和0.32±0.04,骨髓瘤原代细胞中HIF1α、IGF1mRNA表达水平分别由2.02±0.16和1.92±0.13降至0.53±0.04和0.58±0.03,差异均有统计学意义(P值均＜0.05),且呈剂量依赖性;10、20、40 μmol/L罗格列酮均可抑制RPMI8226细胞pAKT、pERK蛋白的表达,然而对骨髓瘤原代细胞T-AKT和T-ERK蛋白表达没有明显影响.结论 罗格列酮可抑制HIF1α、IGF1 mRNA的表达,降低pAKT和pERK蛋白的表达可能是其抑制肿瘤血管形成机制之一.
目的 觀察囉格列酮作用後骨髓瘤細胞HIF1α和IGF1 mRNA錶達水平,探討囉格列酮抑製骨髓瘤血管形成的可能機製.方法 以骨髓瘤細胞繫RPMI 8226細胞及5例初診多髮性骨髓瘤患者骨髓漿細胞(CD138免疫磁珠篩選,作為骨髓瘤原代細胞)為研究對象,採用RT-PCR法檢測用不同濃度(10、20、40 μmol/L)囉格列酮處理前後骨髓瘤細胞HIF1α、IGF1 mRNA錶達水平;採用Westernblot法檢測AKT和ERK蛋白燐痠化和非燐痠化錶達的情況.同時以5例缺鐵性貧血患者的骨髓單箇覈細胞為對照組.結果 RPMI8226細胞和骨髓瘤原代細胞HIF1α和IGF1 mRNA錶達顯著高于對照組.囉格列酮濃度為20 μmol/L處理48 h後,與對照組相比,RPMI 8226細胞中HIF1α、IGF1 mRNA相對錶達水平分彆由1.21±0.08和0.62±0.06降至0.75±0.06和0.32±0.04,骨髓瘤原代細胞中HIF1α、IGF1mRNA錶達水平分彆由2.02±0.16和1.92±0.13降至0.53±0.04和0.58±0.03,差異均有統計學意義(P值均＜0.05),且呈劑量依賴性;10、20、40 μmol/L囉格列酮均可抑製RPMI8226細胞pAKT、pERK蛋白的錶達,然而對骨髓瘤原代細胞T-AKT和T-ERK蛋白錶達沒有明顯影響.結論 囉格列酮可抑製HIF1α、IGF1 mRNA的錶達,降低pAKT和pERK蛋白的錶達可能是其抑製腫瘤血管形成機製之一.
목적 관찰라격렬동작용후골수류세포HIF1α화IGF1 mRNA표체수평,탐토라격렬동억제골수류혈관형성적가능궤제.방법 이골수류세포계RPMI 8226세포급5례초진다발성골수류환자골수장세포(CD138면역자주사선,작위골수류원대세포)위연구대상,채용RT-PCR법검측용불동농도(10、20、40 μmol/L)라격렬동처리전후골수류세포HIF1α、IGF1 mRNA표체수평;채용Westernblot법검측AKT화ERK단백린산화화비린산화표체적정황.동시이5례결철성빈혈환자적골수단개핵세포위대조조.결과 RPMI8226세포화골수류원대세포HIF1α화IGF1 mRNA표체현저고우대조조.라격렬동농도위20 μmol/L처리48 h후,여대조조상비,RPMI 8226세포중HIF1α、IGF1 mRNA상대표체수평분별유1.21±0.08화0.62±0.06강지0.75±0.06화0.32±0.04,골수류원대세포중HIF1α、IGF1mRNA표체수평분별유2.02±0.16화1.92±0.13강지0.53±0.04화0.58±0.03,차이균유통계학의의(P치균＜0.05),차정제량의뢰성;10、20、40 μmol/L라격렬동균가억제RPMI8226세포pAKT、pERK단백적표체,연이대골수류원대세포T-AKT화T-ERK단백표체몰유명현영향.결론 라격렬동가억제HIF1α、IGF1 mRNA적표체,강저pAKT화pERK단백적표체가능시기억제종류혈관형성궤제지일.
Objective To observe the effect of rosiglitazone (RGZ) on the mRNA expression of HIF1α and IGF1 genes in the myeloma cells and explore possible mechanism of angiogenesis inhibition.Methods Human myeloma cell line RPMI 8226 and primary myeloma cells from five patients enriched by using CD 138 immunomagnetic beads were treated with different concentrations (10,20,40 μmol/L)of RGZ.The mRNA expression of HIF1α and IGF1 was analyzed in cells treated with RGZ after 48h by RT-PCR,The levels of phosphorylated AKT and ERK proteins were detected by Western blotting.Bone marrow mononuclear cells from five patients with iron deficiency anemia were regarded as control.Results Higher mRNA expression of HIF1α and IGF1 genes in RPMI8226 and in primary myeloma cells was showed as compared to those in control.Treated with RGZ of 20 μmol/L after 48 h,the mRNA expression of HIF1α (1.21±0.08 vs 0.75±0.06) and IGF1 (0.62±0.06 vs 0.32±0.04) in RPMI8226 cells was declined as compared to those without RGC treatment.The same declination was also seen in in primary myeloma cells (HIF 1 α:2.02±0.16 vs 0.53±0.04; IGF 1:1.92±0.13 vs 0.58±0.03).RGZ could inhibit the expression of pAKT and pERK,nor the total AKT and ERK proteins,in RPMI8226 cells in a dose-dependent manner at the concentration of 10 μmol/L,20 μmol/L,and 40 μmol/L.Conclusions RGZ could inhibit the mRNA expression of HIF 1 α and IGF 1.Inhibition of angiogenesis by RGZ may be associated with down-regulation of pAKT and pERK expression.
