中华血液学杂志
中華血液學雜誌
중화혈액학잡지
Chinese Journal of Hematology
2011年
6期
388-391
,共4页
王季石%柴柏胜%方琴%何颖颖%陈珵%杨畅
王季石%柴柏勝%方琴%何穎穎%陳珵%楊暢
왕계석%시백성%방금%하영영%진정%양창
血红素加氧酶-1%氯高血红素%伊马替尼耐药%K562/A02-IM细胞%细胞凋亡
血紅素加氧酶-1%氯高血紅素%伊馬替尼耐藥%K562/A02-IM細胞%細胞凋亡
혈홍소가양매-1%록고혈홍소%이마체니내약%K562/A02-IM세포%세포조망
Heme oxygenase-1%Hemin%Imatinib resistant%K562/A02-IM cells%Cell apoptosis
目的 通过氯高血红素(Hemin)诱导伊马替尼耐药慢性髓系白血病(CML)细胞株K562/A02-IM 中血红素加氧酶-1(HO-1)基因表达,探讨HO-1基因对伊马替尼耐药CML细胞增殖的影响,为治疗CML多药耐药及新药的开发提供思路和实验依据.方法 采用RT-PCR法检测20例CML伊马替尼耐药患者骨髓细胞中HO-1基因的表达,半定量RT-PCR法和Western blot法分别检测不同剂量Hemin处理K562/A02-IM细胞不同时间后HO-1的表达,通过Annexin V/PI双染色法检测细胞凋亡情况,采用MTT法检测Hemin诱导及锌原卟啉抑制HO-1表达与细胞存活率的关系.结果 RT-PCR结果显示耐药患者骨髓细胞中HO-1基因阳性表达,半定量RT-PCR和Western blot法显示,不同浓度的Hemin(0、10、20及40 μmol/L)处理K562/A02-IM细胞16 h后,HO-1的表达量随Hemin浓度的升高而增加,存在剂量依赖关系,而20 μmol/L Hemin分别处理K562/A02-IM细胞0、8、16、24 h后,HO-1的表达肇在处理16 h组最高.Annexin V/PI法检测显示,0、10、20及40 μmol/L Hemin作用于K562/A02-IM细胞16 h后细胞凋亡率分别为(17.61±0.01)%、(12.13±0.11)%、(7.94±0.03)%和(4.62±0.15)%,其抗凋亡的作用呈剂量依赖性;20 μmol/L Hemin作用K562/A02-IM细胞8、16及24 h的细胞凋亡率分别为(14.72±0.05)%、(8.15±0.07)%和(16.37±0.13)%.MTT法检测显示与对照组相比,Hemin诱导K562/A02-IM细胞HO-1基因表达促进了细胞的增殖,且作用存在剂量依赖关系;而锌原卟啉抑制HO-1的表达,促进了细胞的凋亡(P<0.05).结论 CML伊马替尼耐药患者骨髓细胞HO-1基因阳性表达,HO-1 是一种可诱导表达型基因,具有抗细胞凋亡及促进细胞增殖的作用,抑制HO-1 表达可能成为治疗CML耐药的新方法.
目的 通過氯高血紅素(Hemin)誘導伊馬替尼耐藥慢性髓繫白血病(CML)細胞株K562/A02-IM 中血紅素加氧酶-1(HO-1)基因錶達,探討HO-1基因對伊馬替尼耐藥CML細胞增殖的影響,為治療CML多藥耐藥及新藥的開髮提供思路和實驗依據.方法 採用RT-PCR法檢測20例CML伊馬替尼耐藥患者骨髓細胞中HO-1基因的錶達,半定量RT-PCR法和Western blot法分彆檢測不同劑量Hemin處理K562/A02-IM細胞不同時間後HO-1的錶達,通過Annexin V/PI雙染色法檢測細胞凋亡情況,採用MTT法檢測Hemin誘導及鋅原卟啉抑製HO-1錶達與細胞存活率的關繫.結果 RT-PCR結果顯示耐藥患者骨髓細胞中HO-1基因暘性錶達,半定量RT-PCR和Western blot法顯示,不同濃度的Hemin(0、10、20及40 μmol/L)處理K562/A02-IM細胞16 h後,HO-1的錶達量隨Hemin濃度的升高而增加,存在劑量依賴關繫,而20 μmol/L Hemin分彆處理K562/A02-IM細胞0、8、16、24 h後,HO-1的錶達肇在處理16 h組最高.Annexin V/PI法檢測顯示,0、10、20及40 μmol/L Hemin作用于K562/A02-IM細胞16 h後細胞凋亡率分彆為(17.61±0.01)%、(12.13±0.11)%、(7.94±0.03)%和(4.62±0.15)%,其抗凋亡的作用呈劑量依賴性;20 μmol/L Hemin作用K562/A02-IM細胞8、16及24 h的細胞凋亡率分彆為(14.72±0.05)%、(8.15±0.07)%和(16.37±0.13)%.MTT法檢測顯示與對照組相比,Hemin誘導K562/A02-IM細胞HO-1基因錶達促進瞭細胞的增殖,且作用存在劑量依賴關繫;而鋅原卟啉抑製HO-1的錶達,促進瞭細胞的凋亡(P<0.05).結論 CML伊馬替尼耐藥患者骨髓細胞HO-1基因暘性錶達,HO-1 是一種可誘導錶達型基因,具有抗細胞凋亡及促進細胞增殖的作用,抑製HO-1 錶達可能成為治療CML耐藥的新方法.
목적 통과록고혈홍소(Hemin)유도이마체니내약만성수계백혈병(CML)세포주K562/A02-IM 중혈홍소가양매-1(HO-1)기인표체,탐토HO-1기인대이마체니내약CML세포증식적영향,위치료CML다약내약급신약적개발제공사로화실험의거.방법 채용RT-PCR법검측20례CML이마체니내약환자골수세포중HO-1기인적표체,반정량RT-PCR법화Western blot법분별검측불동제량Hemin처리K562/A02-IM세포불동시간후HO-1적표체,통과Annexin V/PI쌍염색법검측세포조망정황,채용MTT법검측Hemin유도급자원계람억제HO-1표체여세포존활솔적관계.결과 RT-PCR결과현시내약환자골수세포중HO-1기인양성표체,반정량RT-PCR화Western blot법현시,불동농도적Hemin(0、10、20급40 μmol/L)처리K562/A02-IM세포16 h후,HO-1적표체량수Hemin농도적승고이증가,존재제량의뢰관계,이20 μmol/L Hemin분별처리K562/A02-IM세포0、8、16、24 h후,HO-1적표체조재처리16 h조최고.Annexin V/PI법검측현시,0、10、20급40 μmol/L Hemin작용우K562/A02-IM세포16 h후세포조망솔분별위(17.61±0.01)%、(12.13±0.11)%、(7.94±0.03)%화(4.62±0.15)%,기항조망적작용정제량의뢰성;20 μmol/L Hemin작용K562/A02-IM세포8、16급24 h적세포조망솔분별위(14.72±0.05)%、(8.15±0.07)%화(16.37±0.13)%.MTT법검측현시여대조조상비,Hemin유도K562/A02-IM세포HO-1기인표체촉진료세포적증식,차작용존재제량의뢰관계;이자원계람억제HO-1적표체,촉진료세포적조망(P<0.05).결론 CML이마체니내약환자골수세포HO-1기인양성표체,HO-1 시일충가유도표체형기인,구유항세포조망급촉진세포증식적작용,억제HO-1 표체가능성위치료CML내약적신방법.
Objective To investigate the effect of heme oxygenase-1 (HO-1 ) expression on cell growth and apoptosis in imatinib resistant chronic myeloid leukemia (CML) cells ( K562/A02-IM) , and explore the relationship between HO-1 gene and CML. Methods The expression of HO-1 in 20 drug-resistant CML patients was detected by RT-PCR. Different concentrations of hemin were used to induce HO-1 expression of K562/A02-IM, HO-1 expression at different time was detected by RT-PCR and Western blot analysis. Cell apoptosis was detected by Annexin V/PI staining, and MTT assay was used to detect viability of K562/ A02-IM cells after induction or inhibition of HO-1 gene by hemin and zinc protoporphyrin (ZPP). Results RT-PCR showed that HO-1 was expressed in the bone marrow mononuclear cells (BMMNCs). When treated with hemin at different concentrations (0, 10, 20, 40 μmol/L) for 16 h, the expression of HO-1 in K562/ A02-IM was increased in a dose-dependent manner, and peaked at 20 μmol/L of hemin for 16 h. The apoptosis rates were (17.61 ±0.01)%, (12. 13 ±0.11)%, (7.94 ±0.03)% and (4.62 ±0. 15)% at 0,10, 20 and 40 μmol/L of hemin respectively for 16 h and were (14. 7 ± 0.05) % , (8. 1 ± 0. 07) % and (16. 3 ± 0. 13)% at 20 μmol/L of hemin treatment for 8,16, and 24 h respectively. Hemin induced apoptosis of K562/A02-IM cells in a dose-dependent manner. The expression of HO-1 was induced in K562/A02-IM cells in a dose-dependent manner, and the survival of K562/A02-IM cells was significantly increased as compared to that of control group. When HO-1 was inhibited by ZPP, the cells survival was sharply decreased compared to that of the control group (P<0.05). Conclusion HO-1 was expressed in the BMMNCs. It is a kind of molecules whose expression can be induced and can promote the growth of drug-resistant cells. Inhibition of HO-1 expression probably be used for the treatment of drug-resistant CML.
