Salehi Mitra, Piri Hossein, Farasat Alireza, Pakbin Babak, Gheibi Nematollah
Student Research Committee, Qazvin University of Medical Sciences, Qazvin, Iran.
Department of Biochemistry and Genetics, School of Medicine, Qazvin University of Medical Sciences, Qazvin, Iran.
Iran J Basic Med Sci. 2022 Apr;25(4):489-496. doi: 10.22038/IJBMS.2022.60651.13444.
Humic acid (HA) and Fulvic acid (FA) are major members of humic substances, which are extracted from organic sources including soil and peat. The pro-apoptotic and anti-melanogenic effects of HA and FA at the cellular and molecular levels in the A375 human melanoma cell line were examined in this study.
The cytotoxicity effect of HA and FA were evaluated by cell viability assay. Apoptosis and cell cycle were investigated by flow cytometry. Real-time PCR was carried out to measure the expression of , and genes. Moreover, the changes in nanomechanical properties were determined through atomic force microscopy (AFM).
It was found that HA and FA decrease cell viability with an IC value of 50 µg/ml (dose-dependent) for 14 hr, arrested cells in the G0/G1 phase, and increased the sub-G1 phase (induce apoptosis). Based on the AFM analysis, Young's modulus and adhesion force values were increased, also ultrastructural characteristics of cells were changed. Results of Real-time PCR revealed that HA and FA lead to a decrease in the expressions of and genes, and increase the gene expression.
These results exhibited that HA and FA possess pro-apoptotic effects through increasing the expression in A375 cells. These molecular reports were confirmed by cellular nanomechanical assessments using AFM and flow cytometry. In addition, HA and FA inhibited melanogenesis by decreasing the expression of the gene. It is worthwhile to note that, HA and FA can be regarded to design new anti-cancer and anti-melanogenesis products.
腐殖酸（HA）和富里酸（FA）是腐殖质的主要成分，它们从包括土壤和泥炭在内的有机来源中提取。本研究检测了HA和FA在A375人黑色素瘤细胞系中细胞和分子水平上的促凋亡和抗黑色素生成作用。
通过细胞活力测定评估HA和FA的细胞毒性作用。采用流式细胞术研究细胞凋亡和细胞周期。进行实时PCR以测量 、 和 基因的表达。此外，通过原子力显微镜（AFM）测定纳米力学性能的变化。
发现HA和FA在14小时内以50 µg/ml的IC值降低细胞活力（剂量依赖性），使细胞停滞在G0/G1期，并增加亚G1期（诱导凋亡）。基于AFM分析，杨氏模量和粘附力值增加，细胞的超微结构特征也发生了变化。实时PCR结果显示，HA和FA导致 和 基因的表达降低，并增加 基因的表达。
这些结果表明，HA和FA通过增加A375细胞中 的表达具有促凋亡作用。这些分子报告通过使用AFM和流式细胞术的细胞纳米力学评估得到证实。此外，HA和FA通过降低 基因的表达抑制黑色素生成。值得注意的是，HA和FA可被视为设计新的抗癌和抗黑色素生成产品。
