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Sodium Salicylate-induced apoptosis in HL-60 cells and its partial reversing effect on multidrug resistance in HL-60/VCR cells

Author: WanYouFeng
Tutor: ChenXieQun
School: Fourth Military Medical University
Course: Internal Medicine
Keywords: sodium salicylate apoptosis HL-60 cells HL-60/VCR cells Ca2+ multidrug resistance
CLC: R733.7
Type: Master's thesis
Year: 2002
Downloads: 44
Quote: 0
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Abstract


Objective Multidrug resistance(MDR) is the biological basis of refractory leukemia. Searching for an efficient reversing agent of MDR is always important in the clinical research of leukemia. Sodium salicylate(Na-Sal), as a sort of classical nonsteroidal anti-inflamatory drugs(NSAID), rescently showed strong apoptosis-inducing activity against some tumor cells. Because the unability of apoptosis is one of the main mechanisms of MDR in leukemia, we took HL-60 and HL-60/VCR cells as targets to probe into the proapoptotic activity of Na-Sal and its partial reversing effect upon MDR of leukemia. Methods The cytotoxicity of Na-Sal and chemotherapeutical drugs to HL-60 and HL-60/VCR cells was measured by MTT assay; the morphologic changes of target cells were observed by means of light and electronic microscope; the phosphatidylserine exposure was identified with the help of Annexin-V binding assay; the DNA fragmentation was determined by means of agarose gel electrophoresis; the changes of the concentration of intracellular free Ca2+ was analyzed with the help of confocal laser scanning microscope (CLSM); the aneuploid DNA and the concentrationof intracellular daunorubicin(DNR) were analyzed through flow cytometry. Result Na-Sal was able to inhibit the growth of HL-60 cells in a dose- and time-dependent manner. After treatment of HL-60 cells with Na-Sal in a concentration of Smmol-L’^ICso) for lOh 16h, the target cells were characterized morphologically according to condensation of the nuclear chromatin, shrinkage and formation of apoptotic bodies; the phosphatidylserine exposure was highly increased compared with control (0.24 versus 0.03); the DNA from target cells showed typical aneuploid peak by flow cytometry and characteristic "ladder" pattern in agarose gel electrophoresis; and, meanwhile, the concentration of intracellular Ca2+ in target cells was progressively elevated. After exposure of HL-60/VCR cells with non-cytotoxic dose of Na-Sal(lmmol-L"’), the sensitivity of target cells to chemotherapeutical agents was remarkably increased , indicating that the MDR of HL-60/VCR cells was partly reversed. The reversal times varied from 1.01 to 3.26 with a relative reversal efficiency reaching 0.52 % to 73.62 % .With the help of flow cytometry, we found no increase of intracellular concentration of daunorubicin in HL-60/VCR cells during Na-Sal-mediated reverse of MDR. Conclusion (1) Na-Sal is potent in inducing apoptosis of HL-60 cells, which appears to be closely associated with the increase in the concentration of intracellular free Ca2+; (2) Na-Sal can partly reverse MDR of HL-60/VCR cells without changes of the concentration of intracellular DNR, showing that Na-Sal-mediated reverse of MDR in HL-60/VCR cells appears not to affect the pump action of glycoprotein 170(gpl70).

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CLC: > Medicine, health > Oncology > Hematopoietic and lymphoid neoplasms > Leukemia
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