Department of Medical Genetics, Faculty of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
Department of Clinical Biochemistry, Faculty of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
Cell Biochem Biophys. 2020 Dec;78(4):531-539. doi: 10.1007/s12013-020-00936-z. Epub 2020 Aug 14.
Mesenchymal stem cells have been considered as the suitable source for the repair of kidney lesions. The study and identification of novel approaches could improve the efficiency of these cells in the recovery of kidney. In the present study, the effect of HEK 293 conditioned medium (HEK293-CM) was evaluated on the expression of GDNF/RET signaling pathway and their downstream genes in the human adipose-derived mesenchymal stem cells (AD-MSCs). For this purpose, the human AD-MSCs were cultured in the medium containing HEK293-CM. After the RNA extraction and cDNA synthesis, the expression level of GFRA1, GDNF, SPRY1, ETV4, ETV5, and CRLF1 genes were determined by SYBR Green Real time PCR. The obtained results indicated that the GDNF and GFRA1 expression enhanced in the AD-MSCs following treatment with 10% HEK293-CM-5%FBS as compared to the untreated AD-MSCs. These results were consistent with the decreased expression of SPRY1. The significant increased expression of ETV4, ETV5, and CRLF1 genes also showed that HEK293-CM activated the GDNF/RET signaling pathway in the AD-MSCs (P < 0.05). The obtained data suggested that the treatment with HEK293-CM activated the GDNF/RET signaling pathway in the human AD-MSCs.
间充质干细胞被认为是修复肾脏损伤的合适来源。研究和鉴定新方法可以提高这些细胞在肾脏恢复中的效率。在本研究中，评估了 HEK293 条件培养基 (HEK293-CM) 对人脂肪间充质干细胞 (AD-MSCs) 中 GDNF/RET 信号通路及其下游基因表达的影响。为此，将人 AD-MSCs 培养在含有 HEK293-CM 的培养基中。提取 RNA 并合成 cDNA 后，通过 SYBR Green Real time PCR 测定 GFRA1、GDNF、SPRY1、ETV4、ETV5 和 CRLF1 基因的表达水平。结果表明，与未处理的 AD-MSCs 相比，用 10%HEK293-CM-5%FBS 处理后 AD-MSCs 中的 GDNF 和 GFRA1 表达增强。这些结果与 SPRY1 表达下调一致。ETV4、ETV5 和 CRLF1 基因的显著高表达也表明 HEK293-CM 激活了 AD-MSCs 中的 GDNF/RET 信号通路 (P<0.05)。研究数据表明，HEK293-CM 处理激活了人 AD-MSCs 中的 GDNF/RET 信号通路。
