Callaghan R, van Gorkom L C, Epand R M
Department of Biochemistry, McMaster University Health Sciences Centre, Hamilton, Ontario, Canada.
Br J Cancer. 1992 Nov;66(5):781-6. doi: 10.1038/bjc.1992.360.
Cell lines selected (CHRC5) and transfected (LR-73-1A) for multi-drug resistance have total lipid compositions which are indistinguishable between resistant and parental cells. Lipid composition was evaluated by 1H NMR and the total fatty acid content by GLC. No change in surface hydrophobicity, as measured with the fluorescent probe dansyl-PE, was observed as a result of transfection of CHO cells with the mdr1 gene. However, the selected cell line, CHRC5, showed a decreased surface hydrophobicity. This decreased surface hydrophobicity was indicated by an 8 nm increase in the fluorescence emission of dansyl-PE in the CHRC5 cell line compared with the AB1. Both resistant cell lines showed an increase in the polarisation of the fluorescent probe, TMA-DPH in the plasma membranes corresponding to a 14% and a 24% change in fluorescence polarisation for the selected and transfected cell lines, respectively. This is indicative of reduced mobility of the acyl chains in the resistant cell lines. Both the CHRC5 and the transfected cell lines showed almost a 2-fold increase in the initial rate of membrane cycling. The membrane cycling could be inhibited by a known bilayer stabiliser, the N-carbobenzoxy-D-Phe-L-Phe-Gly. These results indicate that the properties of the plasma membrane from resistant cells are altered compared with their parental cell line.
为多药耐药性而选择(CHRC5)和转染(LR-73-1A)的细胞系,其总脂质组成在耐药细胞和亲本细胞之间没有区别。通过1H NMR评估脂质组成,通过GLC评估总脂肪酸含量。用荧光探针丹磺酰磷脂酰乙醇胺(dansyl-PE)测量,未观察到用mdr1基因转染CHO细胞后表面疏水性有变化。然而,所选细胞系CHRC5显示表面疏水性降低。与AB1相比,CHRC5细胞系中丹磺酰磷脂酰乙醇胺的荧光发射增加8 nm,表明表面疏水性降低。两种耐药细胞系的质膜中荧光探针三甲基胺基二苯基己三烯(TMA-DPH)的极化都增加,所选细胞系和转染细胞系的荧光极化分别变化14%和24%。这表明耐药细胞系中酰基链的流动性降低。CHRC5和转染细胞系的膜循环初始速率都几乎增加了2倍。膜循环可被一种已知的双层稳定剂N-苄氧羰基-D-苯丙氨酸-L-苯丙氨酸-甘氨酸抑制。这些结果表明,与亲本细胞系相比,耐药细胞质膜的性质发生了改变。