重组变构人TNF相关促凋亡配体联合三氧化二砷诱导白血病耐药细胞株凋亡

Apoptosis of the Adriamycin-resistant Leukemia Cell Line Induced by the Recombinant Mutant Human TNF-related Apoptosis-inducing Ligand Combined with Arsenic Trioxide

本研究探讨重组变构人TNF相关促凋亡配体(recombinant mutant human TNF-related apoptosis-inducing ligand,rmhTRAIL)联合三氧化二砷(As2O3)对阿霉素诱导的人慢性髓系白血病红白血病变耐药细胞株K562/AO2(mdr-1+)的促凋亡作用。以亲本阿霉素敏感细胞株K562(mdr-1-)为对照,观察rmhTRAIL作用后细胞形态变化,采用MTT法测定rmhTRAIL、As2O3单药及联合作用后细胞增殖抑制率;碘化丙锭染色的流式细胞术(FACSCalibur)定量检测rmhTRAILL、As2O3单药及联合作用后sub-G1峰占检测细胞的百分比(sub-G1%)。结果表明:rmhTRAIL联合As2O3对K562/AO2和K562增殖的抑制作用优于单药,采用国内金(正均)氏公式判断rmhTRAIL和As2O3联合作用具有协同作用;流式细胞术定量检测sub-G1%显示,rmhTRAIL2 000ng/ml与As2O32μmol/L单独作用下K562/AO2组与K562组凋亡率分别为(34.93±0.10)%、(10.53±0.16)%(p<0.01);(5.95±0.07)%、(3.50±0.01)%(p<0.05),联合作用下细胞凋亡率分别为(50.95±0.91)%、(20.75±0.95)%(p<0.05),K562/AO2组凋亡效应强于K562组。结论:rmhTRAIL可诱导K562、K562/AO2细胞凋亡;rmhTRAIL联合凋亡诱导剂As2O3对白血病耐药细胞株K562/AO2和K562细胞株的凋亡具有协同作用;rmhTRAIL、As2O3对K562/AO2的促凋亡作用优于其亲本K562细胞。

This study was aimed to investigate the effect of recombinant mutant human TNF-related apoptosis-inducing ligand （rmhTRAIL） combined with AS2O3 on inducing apoptosis of adriamycin-resistant leukemia cell line K562/AO2 （mdr-1^＋ ）. The mophologic changes of cells treated with rmhTRAIL were observed by inverted microscope, taking adrianycin-sensitve cell line K562 （rndr-1^- ） as control; the inhibitory rate of cell proliferation after being treated with rmhTRAIL, AS2O3 alone or combined was assayed by MTT method; the apoptosis peaks of K562/AO2 and K562 were quantitatively detected by flow cytometry with PI staining after being treated with rmhTRAIL, As2O3 alone or in combination. The results indicated that the inhibition effect of rmhTRAIL and As2O3 in combination on K562/AO2 and K562 cells was higher than that of riTRAIL and AS2O3 alone （p 〈 0. 01 ）, rrnhTRAIL combined with AS2O3 had synargisfic effect in killing K562/AO2 and K562 cells by king＇s formtda. The apoptosis rates of K562/AO2 and K562 cells were 34.93 ±0. 10% and 10.53 ±0. 16% （p 〈0.01 ）, as well as 5.95 ±0. 07%, and 3.50 ±0. 01% （p 〈 0.05）, 50.95 ±0.91% and 20.75 ±0.95% （p 〈0.05） respectively when their cells were treated by rmhTRAIL and AS2O3 alone. Tne apoptosis rate in K562/AO2 group was higher than that in K562 group. It is concluded that rmhTRAIL can induce K562/AO2 and K562 cell apoptosis; rmhTRAIL combined AS2O3 had synergistic effects; the efficacy of on rmhTRAIL or AS2O3 inducing K562/AO2 cell apoptosis is higher than that on their parental cell line K562.