Department of Immunology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran.
Blood Transfusion Research Center, High Institute for Research and Education in Transfusion Medicine, Tehran, Iran.
Iran J Allergy Asthma Immunol. 2021 Apr 17;20(2):221-232.
Conditioned medium (CM) derived from mesenchymal stem cells (MSCs) contains bioactive molecules including microRNAs (miRs) that could be a potential tool for controlling cancer cells' behavior. Due to the properties of CM, this study assesses the effects of miR-34a related MSC-CM on tumor behavior through the evaluation of migration, invasion, apoptosis, and PDL1 expression in breast cancer cell lines. The miR-34a overexpression vector or scramble control was produced using lentiviral vectors, DNA cloning, and the transfection of the HEK-293T cell line. It was then transduced into human adipose-derived mesenchymal stem cells (hAD-MSCs). MSC-CMs were collected and added onto MDA-MB-231 cell lines. The functional evaluations were performed by transwell, wound healing, and Annexin V/PI methods on the treated MDA-MB-231 cell lines. The PDL1 expression was also assessed by Real-time PCR and western blot. The findings of this study showed that ectopic miR‑34a expression was significantly upregulated in manipulated hASC with miR-34a (p<0.0001). Treatment of MDA-MB-231 cell line with miR-34a-hAD-MSC-CM, scramble-hAD-MSC-CM, or hAD-MSC-CM displayed not only a reduction in the number of migrated or invaded cells (p=0.01) but also an increase in the apoptotic cells in the test group (p=0.02) when compared to the control groups. It also showed down-regulation in the gene (p=0.05) and protein expression levels of PDL1 in the test group. The results of the present study showed that simultaneous application of miR-34a and MSC-CM can be considered as a new method for changing the cancerous microenvironment; and therefore, as a potential strategy in breast cancer therapy.
条件培养基（CM）来源于间充质干细胞（MSCs），其中包含生物活性分子，包括 microRNAs（miRs），这可能是控制癌细胞行为的潜在工具。由于 CM 的特性，本研究通过评估乳腺癌细胞系的迁移、侵袭、凋亡和 PDL1 表达，评估 miR-34a 相关 MSC-CM 对肿瘤行为的影响。使用慢病毒载体、DNA 克隆和 HEK-293T 细胞系转染产生 miR-34a 过表达载体或 scramble 对照。然后将其转导到人脂肪来源的间充质干细胞（hAD-MSCs）中。收集 MSC-CM 并添加到 MDA-MB-231 细胞系上。通过 Transwell、划痕愈合和 Annexin V/PI 方法对处理后的 MDA-MB-231 细胞系进行功能评估。还通过 Real-time PCR 和 Western blot 评估 PDL1 表达。本研究的结果表明，外源性 miR-34a 在经 miR-34a 处理的操纵 hASC 中表达显著上调（p<0.0001）。与对照组相比，用 miR-34a-hAD-MSC-CM、 scramble-hAD-MSC-CM 或 hAD-MSC-CM 处理 MDA-MB-231 细胞系不仅减少了迁移或侵袭细胞的数量（p=0.01），而且增加了实验组的凋亡细胞（p=0.02）。还显示实验组 PDL1 的基因（p=0.05）和蛋白表达水平下调。本研究结果表明，同时应用 miR-34a 和 MSC-CM 可以被认为是改变癌症微环境的一种新方法；因此，它可能成为乳腺癌治疗的一种潜在策略。
