Department of Geriatrics/Department of Gastroenterology and Urology Ward II, Hunan Cancer Hospital; Cancer Hospital Affiliated to Xiangya School of Medicine, Central South University, Changsha 410013, China.
Zhong Nan Da Xue Xue Bao Yi Xue Ban. 2020 Dec 28;45(12):1389-1397. doi: 10.11817/j.issn.1672-7347.2020.190728.
To investigate the effect of adriamycin (ADM), idelalisib or ADM and their combination on cell proliferation and intracellular concentration of ADM, and to explore the reversal effect of idelalisib on drug resistance to ADM.
The K562 and K562/ADM cells were respectively treated with ADM and idelalisib at different concentrations. The 50% inhibitory concentration (IC) and drug resistance index (RI) of ADM to the 2 kinds of cells were measured by methyl thiazolyl tetrazolium (MTT) assay. Non-cytotoxic dose (cell inhibition rate <5%) of idelalisib in the 2 kinds of cells was determined. Then the K562 and k562/ADM cells were divided into the following groups: a K562 cells + ADM group, a K562 cells + ADM + idelalisib group, a K562/ADM cells + ADM group, and a K562/ADM cells + ADM + idelalisib group. The survival rates, the intracellular ATP levels, and the relative concentration of intracellular ADM were detected by MTT method, ATP bioluminescence assay (ATP-BLA) and flow cytometry (FCM), respectively.
The cell survival rates were significantly decreased in a dose-dependent manner when the cells were treated with different doses of ADM (0.001-10.000 mg/L ). The IC value of ADM in the K562 and K562/ADM cells were 0.2 mg/L and 1.0 mg/L, respectively. The RI value was 5. The cell survival rates were also significantly decreased in a dose-dependent manner when the cells were treated with different doses of idelalisib (1-50 μmol/L). The non-cytotoxic dose of idelalisib in the K562 and K562/ADM cells were 25 μmol/L and 15 μmol/L, respectively. The cell survival rates in the ADM+ idelalisib group was less than that in the ADM group (<0.05)；while there was no statistical difference between the ADM+ idelalisib group and the ADM group in the K562 cells (>0.05). The intracellular ATP level in the K562 cells was about (91.502±0.479) mmol/L, and that in the K562/ADM cells was about (24.311±0.349) mmol/L. The intracellular ATP level in the ADM+ idelalisib group in the K562/ADM cells was less than that in the ADM group (<0.05); but there was no statistical difference between the ADM + idelalisib group and the ADM group in the K562 cells (>0.05). The absorption of intracellular ADM in the ADM + idelalisib group in the K562/ADM cells was more than that in the ADM group (<0.05); but there was no statistical difference in the K562 cells between the 2 groups (>0.05). The exclusion of intracellular ADM in the ADM + idelalisib group in the K562/ADM cells was less than that in the ADM group (<0.05 or <0.01)；but there was no statistical difference in the K562 cells between the 2 groups (>0.05).
Idelalisib exerts effect on inhibition of the proliferation in myeloid leukemia K562 and K562/ADM cells, which may partially reverse the drug resistance of K562/ADM cells to ADM. The mechanisms for the effect of idelalisib may be related to increasing the accumulation of ADM and inducing the cell apoptosis in the K562 and K562/ADM cells.
研究阿霉素（ADM）、伊达比星或 ADM 及其联合应用对细胞增殖和 ADM 细胞内浓度的影响，并探讨伊达比星对 ADM 耐药的逆转作用。
用不同浓度的 ADM 和伊达比星分别处理 K562 和 K562/ADM 细胞，采用噻唑蓝（MTT）比色法测定 ADM 对 2 种细胞的 50%抑制浓度（IC）和耐药指数（RI）。确定 2 种细胞中无细胞毒性剂量（细胞抑制率<5%）的伊达比星。然后将 K562 和 K562/ADM 细胞分为以下几组：K562 细胞+ADM 组、K562 细胞+ADM+伊达比星组、K562/ADM 细胞+ADM 组和 K562/ADM 细胞+ADM+伊达比星组。采用 MTT 法、ATP 生物发光法（ATP-BLA）和流式细胞术（FCM）分别检测细胞存活率、细胞内 ATP 水平和细胞内 ADM 相对浓度。
不同剂量 ADM（0.001-10.000mg/L）处理细胞时，细胞存活率呈剂量依赖性显著降低。K562 和 K562/ADM 细胞的 ADM IC 值分别为 0.2mg/L 和 1.0mg/L，RI 值为 5。不同剂量伊达比星（1-50μmol/L）处理细胞时，细胞存活率也呈剂量依赖性显著降低。K562 和 K562/ADM 细胞中伊达比星的无细胞毒性剂量分别为 25μmol/L 和 15μmol/L。ADM+伊达比星组细胞存活率小于 ADM 组（<0.05）；而 ADM+伊达比星组与 ADM 组在 K562 细胞中无统计学差异（>0.05）。K562 细胞内 ATP 水平约为（91.502±0.479）mmol/L，K562/ADM 细胞内 ATP 水平约为（24.311±0.349）mmol/L。ADM+伊达比星组 K562/ADM 细胞内 ATP 水平低于 ADM 组（<0.05）；而 ADM+伊达比星组与 ADM 组在 K562 细胞中无统计学差异（>0.05）。ADM+伊达比星组 K562/ADM 细胞内 ADM 摄取量大于 ADM 组（<0.05）；但 2 组在 K562 细胞中无统计学差异（>0.05）。ADM+伊达比星组 K562/ADM 细胞内 ADM 外排量小于 ADM 组（<0.05 或<0.01）；但 2 组在 K562 细胞中无统计学差异（>0.05）。
伊达比星对髓系白血病 K562 和 K562/ADM 细胞的增殖有抑制作用，可能部分逆转 K562/ADM 细胞对 ADM 的耐药性。伊达比星作用的机制可能与增加 ADM 蓄积和诱导 K562 和 K562/ADM 细胞凋亡有关。
