New Flow Cytometric Method for Detection of Minimally Expressed Multidrug Resistance P‐Glycoprotein on Normal and Acute Leukemia Cells Using Biotinylated MRK16 and Streptavidin‐RED670 Conjugate
New Flow Cytometric Method for Detection of Minimally Expressed Multidrug Resistance P‐Glycoprotein on Normal and Acute Leukemia Cells Using Biotinylated MRK16 and Streptavidin‐RED670 Conjugate
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使用生物素化 MRK16 和链霉亲和素-RED670 缀合物检测正常和急性白血病细胞上最低表达的多药耐药性 P-糖蛋白的新流式细胞术方法
DOI：
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发表时间：
1995
期刊：
影响因子：
--
通讯作者：
R. Ohno
中科院分区：
文献类型：
--
作者：
A. Takeshita;K. Shinjo;Kazunori Ohnishl;R. Ohno
为了评估正常和白血病细胞上多药耐药（MDR）的表达，我们通过新设计的流式细胞术方法检测了P-糖蛋白（P-gp），使用了针对P-gp（MRK 16）的生物素化单克隆抗体（mAb）、链霉亲和素-RED 670偶联物（SA-RED 670）和适当的发射滤光片。与标准方法（如使用链霉亲和素-藻红蛋白（SA-PE）结合物）相比，生物素化MRK 16（B-MRK 16）和SA-RED 670的组合产生更高的灵敏度。在K562、K562/ADR、NOMO-1、NOMO-1/ADR和HL 60细胞中检测敏感性，并与mdr-1基因的逆转录聚合酶链反应（RT-PCR）获得的数据进行比较。流式细胞术检测P-gp阳性率分别为10.4%、99.9%、1.4%、90.4%和0%。MDr-1 mRNA在K562/ ADR和NOMO-1/ADR细胞中表达较强，而在NOMO-1和HL-60细胞中不表达。在K562细胞中，mdr-1在40个循环的PCR后被发现，而不是25个循环。这些数据与来自流式细胞术的数据良好相关。然后，我们研究了正常外周血细胞和急性白血病细胞上的P-gp表达。P-gp在外周血淋巴细胞、单核细胞和粒细胞上几乎不表达。5例急性早幼粒细胞白血病（APL）患者的原始细胞中也有少量表达，表达率为0.2 ~ 10.6%（4.6±3.9%）。另外10例急性髓细胞白血病（AMD）和5例急性淋巴细胞白血病（ALL）在诊断时表达P-gp，范围分别为8.5%-34.5%（16.9± 11.8%）和2.3%-45.6%（24.0±17.8%）。所有9例复发或难治性病例均表达P-gp，范围为21.1%至99.8%（52.2±29.9%）。APL、CD 34阳性、复发和难治性病例之间存在显著差异（分别为P=0.0006、0.0007和0.0088）。这些结果表明，这种流式细胞术分析是有用的临床MDR状态的评估，并可以识别一组耐药白血病患者。
To evaluate the expression of multidrug resistance (MDR) on normal and leukemia cells, we examined P‐glycoprotein (P‐gp) by a newly devised flow cytometric method, utilizing a biotinylated monoclonal antibody (mAb) against P‐gp (MRK16), a streptavidin‐RED670 conjugate (SA‐RED670) and appropriate emission filters. The combination of biotinylated MRK16 (b‐MRK16) and SA‐RED670 resulted in higher sensitivity as compared with standard methods such as the use of streptavidin‐phycoerythrin (SA‐PE) conjugate. The sensitivity was examined in K562, K562/ADR, NOMO‐1, NOMO‐1/ADR and HL60 cells, and compared with the data obtained from reverse transcription polymerase chain reaction (RT‐PCR) of mdr‐1 gene. P‐gp positively on flow cytometry was 10.4%, 99.9%, 1.4%, 90.4% and 0%, respectively. Mdr‐1 mRNA was well expressed in K562/ ADR and NOMO‐1/ADR cells, hut not in NOMO‐1 and HL60 cells. In K562 cells, mdr‐1 was found after 40 cycles of PCR, but not 25 cycles. These data are well correlated with those from the flow cytometry. We then studied the P‐gp expression on normal peripheral blood cells and acute leukemia cells. P‐gp was little expressed on peripheral lymphocytes, monocytes and granulocytes. It was also little expressed on blast cells from 5 patients with acute promyelocytic leukemia (APL) at diagnosis, ranging from 0.2 to 10.6% (4.6±3.9%). Ten other acute myeloid leukemia (AMD and 5 acute lymphocytic leukemia (ALL) expressed P‐gp at diagnosis, ranging from 8.5% to 34.5% (16.9± 11.8%) and from 2.3% to 45.6% (24.0±17.8%), respectively. All 9 relapsed or refractory cases expressed P‐gp, ranging from 21.1% to 99.8% (52.2±29.9%). Significant differences were found in APL, CD34‐positive and relapse and refractory cases (P=0.0006, 0.0007 and 0.0088, respectively). These results indicate that this flow cytometric analysis is useful for the evaluation of clinical MDR status and can identify a group of patients with resistant leukemia.
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影响因子：
20.3
作者：
D. Drach;Shourong Zhao;J. Drach;R. Mahadevia;Claus Gattringer;Heinz Huber;M. Andreeff
通讯作者：
D. Drach;Shourong Zhao;J. Drach;R. Mahadevia;Claus Gattringer;Heinz Huber;M. Andreeff
人外周血淋巴细胞中多药耐药P-糖蛋白的表达和活性。
影响因子：
20.3
作者：
Chaudhary,PM;Mechetner,EB;Roninson,IB
通讯作者：
Roninson,IB
影响因子：
10.3
作者：
Jonker, JW;Smit, JW;Schinkel, AH
通讯作者：
Schinkel, AH
通过 MRK-16 单克隆抗体的表位作图确定 P-糖蛋白的拓扑结构。
DOI：
--
发表时间：
1993
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Georges,E;Tsuruo,T;Ling,V
通讯作者：
Ling,V
正常志愿者循环血细胞中 P-糖蛋白的表达和功能。
影响因子：
20.3
作者：
Klimecki,WT;Futscher,BW;Grogan,TM;Dalton,WS
通讯作者：
Dalton,WS