中国药理学通报
中國藥理學通報
중국약이학통보
CHINESE PHARMACOLOGICAL BULLETIN
2014年
9期
1214-1218,1219
,共6页
李真真%张晓龙%杨雨琪%张晴%袁向飞%范冬梅
李真真%張曉龍%楊雨琪%張晴%袁嚮飛%範鼕梅
리진진%장효룡%양우기%장청%원향비%범동매
ABCB5%P-gp%急性髓系白血病%siABCB5%多药耐药%复发难治
ABCB5%P-gp%急性髓繫白血病%siABCB5%多藥耐藥%複髮難治
ABCB5%P-gp%급성수계백혈병%siABCB5%다약내약%복발난치
ABCB5%P-gp%AML%siABCB5%MDR%relapse/refractory
目的：探讨ABCB5和MDR1在急性髓系白血病( acute myeloid leukemia, AML)干祖细胞系KG1a和来源于AML病人标本中的表达及对白血病耐药的影响。方法 FACS和Western blot的方法检测KG1a细胞表面ABCB5及MDR1蛋白表达产物P-gp的表达水平；利用lipo2000转染siRNA的方法瞬时干扰 ABCB5的表达；FACS 检测细胞内 Rhoda-mine123的蓄积；MTT法确定维拉帕米( verapamil, Vera)对KG1a的无毒剂量,并检测KG1a、siABCB5-KG1a及联用无毒剂量维拉帕米的KG1a对阿霉素( adriamycin, ADR)的敏感性；Real time-PCR的方法检测 AML病人标本中 ABCB5及MDR1的表达水平。结果 KG1a高表达ABCB5和P-gp,P-gp表达量高于ABCB5；siABCB5瞬时干扰可以降低KG1a中ABCB5的表达水平,进而增加 KG1a 细胞中 Rhodamine123的蓄积；siABCB5-KG1a对阿霉素的敏感性提高8.6倍；而联用无毒剂量维拉帕米使KG1a对阿霉素的敏感性提高67.4倍。 ABCB5在71例AML临床标本中的表达水平明显高于健康组,其中在38例复发或难治型病例中的表达水平明显高于其余33例化疗敏感型病例( P <0.01),且 ABCB5的mRNA表达水平与MDR1呈正相关。结论 ABCB5与已知的P-gp相同,也参与介导AML的多药耐药,这为复发及难治的AML病例的临床治疗提供新的潜在靶点。
目的：探討ABCB5和MDR1在急性髓繫白血病( acute myeloid leukemia, AML)榦祖細胞繫KG1a和來源于AML病人標本中的錶達及對白血病耐藥的影響。方法 FACS和Western blot的方法檢測KG1a細胞錶麵ABCB5及MDR1蛋白錶達產物P-gp的錶達水平；利用lipo2000轉染siRNA的方法瞬時榦擾 ABCB5的錶達；FACS 檢測細胞內 Rhoda-mine123的蓄積；MTT法確定維拉帕米( verapamil, Vera)對KG1a的無毒劑量,併檢測KG1a、siABCB5-KG1a及聯用無毒劑量維拉帕米的KG1a對阿黴素( adriamycin, ADR)的敏感性；Real time-PCR的方法檢測 AML病人標本中 ABCB5及MDR1的錶達水平。結果 KG1a高錶達ABCB5和P-gp,P-gp錶達量高于ABCB5；siABCB5瞬時榦擾可以降低KG1a中ABCB5的錶達水平,進而增加 KG1a 細胞中 Rhodamine123的蓄積；siABCB5-KG1a對阿黴素的敏感性提高8.6倍；而聯用無毒劑量維拉帕米使KG1a對阿黴素的敏感性提高67.4倍。 ABCB5在71例AML臨床標本中的錶達水平明顯高于健康組,其中在38例複髮或難治型病例中的錶達水平明顯高于其餘33例化療敏感型病例( P <0.01),且 ABCB5的mRNA錶達水平與MDR1呈正相關。結論 ABCB5與已知的P-gp相同,也參與介導AML的多藥耐藥,這為複髮及難治的AML病例的臨床治療提供新的潛在靶點。
목적：탐토ABCB5화MDR1재급성수계백혈병( acute myeloid leukemia, AML)간조세포계KG1a화래원우AML병인표본중적표체급대백혈병내약적영향。방법 FACS화Western blot적방법검측KG1a세포표면ABCB5급MDR1단백표체산물P-gp적표체수평；이용lipo2000전염siRNA적방법순시간우 ABCB5적표체；FACS 검측세포내 Rhoda-mine123적축적；MTT법학정유랍파미( verapamil, Vera)대KG1a적무독제량,병검측KG1a、siABCB5-KG1a급련용무독제량유랍파미적KG1a대아매소( adriamycin, ADR)적민감성；Real time-PCR적방법검측 AML병인표본중 ABCB5급MDR1적표체수평。결과 KG1a고표체ABCB5화P-gp,P-gp표체량고우ABCB5；siABCB5순시간우가이강저KG1a중ABCB5적표체수평,진이증가 KG1a 세포중 Rhodamine123적축적；siABCB5-KG1a대아매소적민감성제고8.6배；이련용무독제량유랍파미사KG1a대아매소적민감성제고67.4배。 ABCB5재71례AML림상표본중적표체수평명현고우건강조,기중재38례복발혹난치형병례중적표체수평명현고우기여33례화료민감형병례( P <0.01),차 ABCB5적mRNA표체수평여MDR1정정상관。결론 ABCB5여이지적P-gp상동,야삼여개도AML적다약내약,저위복발급난치적AML병례적림상치료제공신적잠재파점。
Aim To investigate the expression of AB-CB5 and MDR1 in the cell line KG1 a and samples from acute myeloid leukemia ( AML) and their effects on multidrug resistance. Methods The expression of ABCB5 and P-gp ( the expressed product of MDR1 ) in KG1 a cells were detected by flow cytometry as well as Western blot analysis; KG1 a cells were transfected with the specific siRNA of ABCB5 using lipo2000 to reduce the expression of ABCB5; intracellular rhoda-mine123 was measured by flow cytometry;cell viability was detected by MTT; the expressions of ABCB5 and MDR1 in samples from AML were detected by real time PCR. Results ABCB5 and P-gp were overexpressed in KG1 a;the specific siRNA of ABCB5 transiently in-hibited the expression of ABCB5 in KG1 a; the siAB-CB5-KG1 a cells increased the intracellular rhodamine 123 and have been more sensitive to adriamycin com-pared with the parent KG1a. ABCB5 gene expression in samples from AML was higher than healthy people. Further, the expression of ABCB5 in 38 relapse or re-fractory AML significantly exceeded the 33 drug sensi-tive. And we found a significant positive correlation between ABCB5 expression and MDR1 gene expression in the 38 patients with relapse or refractory AML. Conclusion ABCB5 , as well as P-gp contributes to mediate multidrug resistance of AML, which provides a novel target for the therapy of relapse or refractory AML.
