Department of Medical Oncology, Henan Tumor Hospital, Zhengzhou, Henan 450000, China.
Chin Med J (Engl). 2013 Mar;126(5):925-9.
Multidrug resistance (MDR) is a main reason for paclitaxel (TAX) treatment failure. Indirubin-3'-monoxime (IRO) and Matrine are traditional Chinese medicines, which may reverse the resistance of tumor cells to some chemotherapy drugs, but the relationship between paclitaxel resistance and Matrine is still unclear. The aim of this study was to explore the potential molecular mechanism of IRO and Matrine in reversal of TAX resistance.
In this study, MTT assay was used to measure the non-cytotoxic dosage of IRO and Matrine on NCI-H520/TAX25 cells and determine the reversal extent of TAX resistance under non-toxic doses. In addition, RT-PCR and Western blotting were used to evaluate the mRNA expression and the protein level of survivin, Oct-4, and Sox-2 in NCI-H520/TAX25 cells using semi-quantitative methods.
There was no obvious inhibition on sensitive cell strains and drug-resistant strains, when the final concentration was at lest 4 µmol/L for IRO and 100 µmol/L for Matrine. So 4 µmol/L of IRO and 100 µmol/L of Matrine were considered as the reversal dosage. When 4 µmol/L of IRO or 100 µmol/L of Matrine were used together with TAX, the sensitivity to TAX increased evidently in NCI-H520/TAX2 cells; the reversal rate of IRO and Matrine was about 1.92 (43.56/22.6 nmol/L) and 1.74 (43.56/25.0 nmol/L), respectively. The mRNA expression and the protein level of survivin, Oct-4, and Sox-2 in NCI-H520/TAX25 decreased significantly (P < 0.05) after addition of IRO or Matrine in TAX treatment, compared to that of TAX treatment alone.
The decrease in both mRNA expression and protein level of survivin, Oct-4, and Sox-2 might be the molecular mechanism, by which IRO and Matrine mediate the reversal of TAX resistance.
多药耐药（MDR）是紫杉醇（TAX）治疗失败的主要原因。靛玉红-3'-单肟（IRO）和苦参碱是传统中药，可能逆转肿瘤细胞对某些化疗药物的耐药性，但紫杉醇耐药与苦参碱的关系尚不清楚。本研究旨在探讨 IRO 和苦参碱逆转 TAX 耐药的潜在分子机制。
本研究采用 MTT 法测定 IRO 和苦参碱对 NCI-H520/TAX25 细胞的非细胞毒性剂量，并在无毒剂量下确定 TAX 耐药的逆转程度。此外，采用半定量 RT-PCR 和 Western blot 法检测 NCI-H520/TAX25 细胞中 survivin、Oct-4 和 Sox-2 的 mRNA 表达和蛋白水平。
IRO 终浓度至少为 4 μmol/L，苦参碱终浓度为 100 μmol/L 时，对敏感细胞株和耐药细胞株均无明显抑制作用。因此，4 μmol/L 的 IRO 和 100 μmol/L 的苦参碱被认为是逆转剂量。当 4 μmol/L 的 IRO 或 100 μmol/L 的苦参碱与 TAX 一起使用时，NCI-H520/TAX2 细胞对 TAX 的敏感性明显增加；IRO 和苦参碱的逆转率分别约为 1.92（43.56/22.6 nmol/L）和 1.74（43.56/25.0 nmol/L）。与 TAX 单独处理相比，加入 IRO 或苦参碱后，NCI-H520/TAX25 细胞中 survivin、Oct-4 和 Sox-2 的 mRNA 表达和蛋白水平均显著降低（P<0.05）。
IRO 和苦参碱介导 TAX 耐药逆转的分子机制可能是 survivin、Oct-4 和 Sox-2 的 mRNA 表达和蛋白水平降低。
