中华医学杂志
中華醫學雜誌
중화의학잡지
National Medical Journal of China
2011年
26期
1856-1860
,共5页
江庭秀%顾伟英%邱国强%陈子兴%王志林%吴浩清%华晓莹%贺白%吴炜%谢晓宝%曹祥山
江庭秀%顧偉英%邱國彊%陳子興%王誌林%吳浩清%華曉瑩%賀白%吳煒%謝曉寶%曹祥山
강정수%고위영%구국강%진자흥%왕지림%오호청%화효형%하백%오위%사효보%조상산
白血病,早幼粒细胞,急性%维甲酸%辛伐他汀%WT1基因%hDMP1基因
白血病,早幼粒細胞,急性%維甲痠%辛伐他汀%WT1基因%hDMP1基因
백혈병,조유립세포,급성%유갑산%신벌타정%WT1기인%hDMP1기인
Leukemia,promyelocytic,acute%Tretinoin%Simvastatin%Wilms' tumor gene%Human cyclin-dependent myb-like protein 1 gene
目的 探讨羟甲基戊二酸单酰辅酶A(HMG-CoA)还原酶的抑制剂辛伐他汀(SV)联合全反式维甲酸(ATRA)对人早幼粒细胞白瓶病NB4细胞株增殖、分化与凋亡的影响,以及肾母细胞瘤(WT1)基因/转录因子人细胞周期调节蛋白依赖性myb样蛋白1(hDMP1)基因表达变化.方法 以不同浓度SV联合ATRA处理NB4细胞,取对数生长期细胞进行实验.各组细胞分别进行细胞形态观察;四甲基偶氮唑盐(MTT)法观察细胞增殖能力;流式细胞仪测定NB4细胞分化指标CD11b和细胞凋亡指标膜联蛋白(Annexin-V)/碘化丙啶(PI)变化;实时(Real-time)RT-PCR测定WT1/hDMP1基因表达变化.结果 15、10、5 μmol/L SV单独处理NB4细胞,随着培养时间延长,细胞抑制率增加(F=7.15,P=0.000),CD11b的表达水平逐渐升高(F=3.41,P=0.014),Annexin-V表达水平逐渐增高(F=43.38,P=0.000),WT1基因表达水平逐渐降低(F=5.35,P=0.001),同时伴随着hDMP1基因表达水平的增加(F=22.61,P=0.000),其中以15 μmol/L SV组NB4细胞变化最明显.15 μmol/L SV联合ATRA处理NB4细胞培养72 h CD11b(89.46%±9.13%)和hDMP1(626.9±56.9)表达水平明显高于ATRA(71.27%±7.27%和421.8±38.3)和SV(62.41%±6.37%和241.4±21.9)单药处理(均P<0.05),两药合用具有明显交互作用(F=4.09,P=0.025和F=29.58,P=0.000).联合用药对NB4细胞抑制率和Annexin-V表达水平差异无统计学意义.结论 SV以剂量依赖方式体外抑制NB4细胞生长,诱导NB4细胞的分化,促进凋亡,降低WT1表达,升高hDMP1表达,提示SV具有协同治疗急性早幼粒细胞白血病的潜能.
目的 探討羥甲基戊二痠單酰輔酶A(HMG-CoA)還原酶的抑製劑辛伐他汀(SV)聯閤全反式維甲痠(ATRA)對人早幼粒細胞白瓶病NB4細胞株增殖、分化與凋亡的影響,以及腎母細胞瘤(WT1)基因/轉錄因子人細胞週期調節蛋白依賴性myb樣蛋白1(hDMP1)基因錶達變化.方法 以不同濃度SV聯閤ATRA處理NB4細胞,取對數生長期細胞進行實驗.各組細胞分彆進行細胞形態觀察;四甲基偶氮唑鹽(MTT)法觀察細胞增殖能力;流式細胞儀測定NB4細胞分化指標CD11b和細胞凋亡指標膜聯蛋白(Annexin-V)/碘化丙啶(PI)變化;實時(Real-time)RT-PCR測定WT1/hDMP1基因錶達變化.結果 15、10、5 μmol/L SV單獨處理NB4細胞,隨著培養時間延長,細胞抑製率增加(F=7.15,P=0.000),CD11b的錶達水平逐漸升高(F=3.41,P=0.014),Annexin-V錶達水平逐漸增高(F=43.38,P=0.000),WT1基因錶達水平逐漸降低(F=5.35,P=0.001),同時伴隨著hDMP1基因錶達水平的增加(F=22.61,P=0.000),其中以15 μmol/L SV組NB4細胞變化最明顯.15 μmol/L SV聯閤ATRA處理NB4細胞培養72 h CD11b(89.46%±9.13%)和hDMP1(626.9±56.9)錶達水平明顯高于ATRA(71.27%±7.27%和421.8±38.3)和SV(62.41%±6.37%和241.4±21.9)單藥處理(均P<0.05),兩藥閤用具有明顯交互作用(F=4.09,P=0.025和F=29.58,P=0.000).聯閤用藥對NB4細胞抑製率和Annexin-V錶達水平差異無統計學意義.結論 SV以劑量依賴方式體外抑製NB4細胞生長,誘導NB4細胞的分化,促進凋亡,降低WT1錶達,升高hDMP1錶達,提示SV具有協同治療急性早幼粒細胞白血病的潛能.
목적 탐토간갑기무이산단선보매A(HMG-CoA)환원매적억제제신벌타정(SV)연합전반식유갑산(ATRA)대인조유립세포백병병NB4세포주증식、분화여조망적영향,이급신모세포류(WT1)기인/전록인자인세포주기조절단백의뢰성myb양단백1(hDMP1)기인표체변화.방법 이불동농도SV연합ATRA처리NB4세포,취대수생장기세포진행실험.각조세포분별진행세포형태관찰;사갑기우담서염(MTT)법관찰세포증식능력;류식세포의측정NB4세포분화지표CD11b화세포조망지표막련단백(Annexin-V)/전화병정(PI)변화;실시(Real-time)RT-PCR측정WT1/hDMP1기인표체변화.결과 15、10、5 μmol/L SV단독처리NB4세포,수착배양시간연장,세포억제솔증가(F=7.15,P=0.000),CD11b적표체수평축점승고(F=3.41,P=0.014),Annexin-V표체수평축점증고(F=43.38,P=0.000),WT1기인표체수평축점강저(F=5.35,P=0.001),동시반수착hDMP1기인표체수평적증가(F=22.61,P=0.000),기중이15 μmol/L SV조NB4세포변화최명현.15 μmol/L SV연합ATRA처리NB4세포배양72 h CD11b(89.46%±9.13%)화hDMP1(626.9±56.9)표체수평명현고우ATRA(71.27%±7.27%화421.8±38.3)화SV(62.41%±6.37%화241.4±21.9)단약처리(균P<0.05),량약합용구유명현교호작용(F=4.09,P=0.025화F=29.58,P=0.000).연합용약대NB4세포억제솔화Annexin-V표체수평차이무통계학의의.결론 SV이제량의뢰방식체외억제NB4세포생장,유도NB4세포적분화,촉진조망,강저WT1표체,승고hDMP1표체,제시SV구유협동치료급성조유립세포백혈병적잠능.
Objective To investigate the effects of simvastatin (SV) plus all-trans retinoic acid ( ATRA) on the proliferation, differentiation, apoptosis and WT1/hDMP1 gene expression profiles of human promyelocytic leukemia cell line NB4. Methods The NEW cell was incubated with simvastatin and ATRA alone or in combination. And the NB4 cell without any treatment was adopted as a normal control. The cells of different groups were collected at24, 48 and 72 h post-incubation. Their morphological changes were observed after Wright staining. The method of MTT was employed to assay the growth inhibition rate and flow cytometry was used to detect the early-stage ratios of apoptosis and cell necrosis. Real-time quantitative reverse transcriptase-polymerase chain reaction ( RT-PCR) was used to detect the WT1/hDMP1 gene expression levels. Results The cell inhibition rates increased gradually (F = 7. 15, P = 0.000) at 15, 10 and 5 μmol/L SV respectively. And so did the expression levels of CD11b ( F= 3. 41, P = 0. 014) and Annexin-V ( F = 43. 38, P = 0. 000). However the expression levels of WT1 decreased gradually ( F =5. 35, P = 0. 001) reversely with the elevated levels of hDMP1 ( F= 22. 61, P = 0. 000 ). Furthermore the NB4 cell exhibited the most significant changes at 15 μmol/L SV. After a 72-hour incubation, the expression levels of GD11b(89. 46% ±9. 13% )and hDMP1(626. 9 ±56.9)in NB4 cells at 15 μmol/L SV plus 0. 5 μmol/L ATRA were significantly higher than those with ATRA(71. 27% ±7. 27% , P =0. 000 and 421. 8 ±38. 3, P =0.003 in each) and SV alone(62.41% ±6.37%, P =0.003 and 241. 4 ±21. 9, P =0.003 in each). A combination of 15 μmol/L SV with 0.5 μmol/L ATRA displayed obvious interactions with the expressions of CD11b and hDMPl (F = 4. 09, P = 0. 025 and F = 29. 58, P = 0. 000 in each). And there was no significant interaction for cell inhibition rates and Annexin-V expression. Conclusion Simvastatin in vitro inhibits the proliferation of NB4 cell, induces its differentiation and promotes its apoptosis. And the lowered expression of WT1 has a dose-dependent correlation with the elevated expression of hDMP1. It indicates that simvastatin has the synergistic in vitro anti-promyelocytic potency.
