Department of Genetics, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, No. 2, Hafez St., Banihashem St., Resalat Highway, P.O.Box:16635-148, Tehran, Iran.
Department of Genetics, Faculty of Biological Science, Shahid Beheshti University, Tehran, Iran.
Mol Biol Rep. 2020 Nov;47(11):8605-8614. doi: 10.1007/s11033-020-05903-8. Epub 2020 Oct 15.
Chromosomal abnormalities, as a frequent phenomenon in cultured embryonic stem cells (ESCs), is a major obstacle in the ESC application in regenerative medicine. Recent studies showed that aneuploid embryonic stem cells of humans and mice are more vulnerable to anticancer drugs, compared with normal cells. The aim of the current study was to evaluate effects of three anticancer drugs, paclitaxel, lapatinib and bortezomib, on mouse embryonic stem cells (mESCs) as a suitable and available model. To assess in vitro cell toxicity, two mESC lines were treated with the aforementioned drugs. Using G-band karyotyping and micronucleus assay, the effect of anticancer drugs in terms of reduction of chromosomal instability in the mESCs was evaluated in control and treatment groups. Also, apoptosis rate of both groups was estimated by FITC-Annexin V/Propidium Iodide (PI) double staining. In addition, the effect of these three drugs in maintaining the pluripotency was assessed through alkaline phosphatase assay and quantification of the expression of three key pluripotency genes, Nanog, Pou5f1 and Sox-2 was performed using Real Time PCR. The rate of numerical abnormalities after treatment with paclitaxel and lapatinib was lower than the control group. The expression level of pluripotency genes exhibited no significant difference between control and treatment groups. Administration of paclitaxel and lapatinib to the mESCs culture at an appropriate dose and in a timely manner could decrease chromosome stability without affecting pluripotency.
染色体异常是培养胚胎干细胞（ESC）中常见的现象，是 ESC 在再生医学应用中的主要障碍。最近的研究表明，与正常细胞相比，人类和小鼠的非整倍体胚胎干细胞对抗癌药物更敏感。本研究旨在评估三种抗癌药物紫杉醇、拉帕替尼和硼替佐米对小鼠胚胎干细胞（mESC）的影响，将其作为合适的、可用的模型。为了评估体外细胞毒性，用上述药物处理两种 mESC 系。通过 G 带核型分析和微核试验，评估抗癌药物对 mESC 中染色体不稳定性降低的影响，分别在对照组和治疗组中进行。此外，通过 FITC-Annexin V/PI 双染色法估计两组的凋亡率。此外，还通过碱性磷酸酶试验评估这三种药物对维持多能性的影响，并通过实时 PCR 定量检测三个关键多能性基因 Nanog、Pou5f1 和 Sox-2 的表达水平来评估。紫杉醇和拉帕替尼治疗后的数值异常率低于对照组。多能性基因的表达水平在对照组和治疗组之间没有显著差异。在适当的剂量和时间内将紫杉醇和拉帕替尼施用于 mESC 培养物中，可以降低染色体稳定性，而不影响多能性。
