Kunte Mugdha, Desai Krutika
Department of Biological Sciences, NMIMS University, Vile Parle (W), Mumbai, India.
Department of Microbiology, Mithibai College, Vile Parle (W), Mumbai, India.
Cell J. 2018 Jul;20(2):211-219. doi: 10.22074/cellj.2018.5277. Epub 2018 Mar 18.
Considering the bioactivities exhibited by microalgae, the effect of protein extract of Chlorella minutissimma (CP extract) was investigated on the expression of human matrix metalloproteinases-1 (MMP-1) in the breast cancer cell line MDA-MB231, and that of MMP-2 and -9 in hepatocellular cancer cell line HepG2 at different expression levels. The study aimed identification and analysis of inhibitory activity of microalgal components extracted from Chlorella minutissima against human MMPs.
In this experimental study, we analysed the effect of Chlorella extracts on MMP-1, -2, and -9 expression at various levels. Gelatin zymography was performed to study the inhibitory effect of Chlorella exracts on human gelatinases at the activity level, followed by western blotting to analyse the expression of all three MMPs at the protein level. The similar effect at the mRNA level along with the probable mechanism underlying inhibition of MMPs was assessed using real-time polymerase chain reaction (PCR).
The results reveal that the treatment with CP extract decreased the mRNA expression of MMP-1, MMP-2, and MMP-9 by 0.26-, 0.29-, and 0.40-fold, respectively, at 20 μg/ml concentration as well as inhibited the activity of MMP-2 and MMP-9 by 37.56 and 42.64%, respectively, at 15 μg/ml concentration. Additionally, upregulated mRNA expression of tissue inhibitor of metalloproteinases-3 (TIMP-3) by 1.68-fold was seen in HepG2 cells at 20 μg/ml concentration treatment group. However, CP extract did not induce any change in the mRNA expression of the TIMP-1, -2 and -4 in HepG2 and TIMP-1, -2, -3 and -4 in MDA-MB231 cells. Activator protein-1 (AP-1)-dependent c-Jun-mediated transcriptional regulation of MMP-1, -2, and -9 was also studied to elucidate the appropriate mechanism involved in the inhibition of MMPs.
The CP extract successfully inhibited MMP-1, -2, and -9 at different expression levels through TIMP-3 upregulation and c-Jun downregulation.
鉴于微藻所展现的生物活性，研究了极小绿藻蛋白提取物（CP提取物）对乳腺癌细胞系MDA-MB231中人基质金属蛋白酶-1（MMP-1）表达的影响，以及对不同表达水平的肝癌细胞系HepG2中MMP-2和MMP-9表达的影响。该研究旨在鉴定和分析从小球藻中提取的微藻成分对人基质金属蛋白酶的抑制活性。
在本实验研究中，我们分析了小球藻提取物在不同水平对MMP-1、MMP-2和MMP-9表达的影响。进行明胶酶谱分析以研究小球藻提取物在活性水平对人明胶酶的抑制作用，随后进行蛋白质印迹分析以在蛋白质水平分析所有三种基质金属蛋白酶的表达。使用实时聚合酶链反应（PCR）评估在mRNA水平的类似作用以及基质金属蛋白酶抑制的潜在机制。
结果显示，在20μg/ml浓度下，CP提取物处理分别使MMP-1、MMP-2和MMP-9的mRNA表达降低0.26倍、0.29倍和0.40倍，并且在15μg/ml浓度下分别使MMP-2和MMP-9的活性抑制37.56%和42.64%。此外，在20μg/ml浓度处理组的HepG2细胞中，金属蛋白酶组织抑制剂-3（TIMP-3）的mRNA表达上调了1.68倍。然而，CP提取物在HepG2细胞中未诱导TIMP-1、TIMP-2和TIMP-4的mRNA表达发生任何变化，在MDA-MB231细胞中也未诱导TIMP-1、TIMP-2、TIMP-3和TIMP-4的mRNA表达发生任何变化。还研究了激活蛋白-1（AP-1）依赖性c-Jun介导的MMP-1、MMP-2和MMP-9的转录调控，以阐明参与基质金属蛋白酶抑制的适当机制。
CP提取物通过上调TIMP-3和下调c-Jun成功地在不同表达水平抑制了MMP-1、MMP-2和MMP-9。
