Botany and Microbiology Department, Faculty of Science, Benha University, Egypt.
Department of Molecular Biology, Genetic Engineering and Biotechnology Research Institute, University of Sadat City, Egypt.
Asian Pac J Cancer Prev. 2024 Oct 1;25(10):3551-3558. doi: 10.31557/APJCP.2024.25.10.3551.
Bidens pilosa and Trianthema portulacastrum are noteworthy weeds with a series of bioactive flavonoid constituents, hence, they can be utilized as potential health supplements and readily available sources of natural antioxidants, as well as effective constituents in medicinal applications. The current study aims to assess the anti-proliferative activity of B. pilosa and T. portulacastrum extracts using the HepG2 cell line. Methods The prepared extracts were evaluated for their cytotoxic influence and their potential CC50 in HepG2 cell lines and normal hepatocytes using the MTT assay. Using quantitative real-time polymerase chain reaction (qRT-PCR), the relative gene expression of Raf-1, MEK-1, LC3B, and Atg12 was quantified in treated cells to detect the expression levels of cell proliferation factors and autophagy-related genes. The quantification analysis of the released interleukin-1beta (IL-1β) and interleukin-1alpha (IL-1α) was also done using an ELISA assay.
The activities of B. pilosa extract showed an anti-proliferative influence on HepG2 cell lines upon treatment as compared to normal cells. It was assessed for cytotoxicity using molecular studies against both Raf-1 and MEK-1 as proposed anticancer mechanisms and showed promising inhibitory activity against Raf-1 and MEK-1 gene expression. Likewise, the reduction of autophagy-related genes, Atg12 and LC3B, in HepG2 cells pre-treated with B. pilosa extract, further confirmed its influence in the induction of programmed cell death (PCD). The ELISA assay revealed a substantial elevation of the pro-inflammatory cytokines IL-1α and IL-1β upon treatment.
This study found that B. pilosa extract, without any detectable cytotoxic effects, had potential inhibitory activities against both Raf-1 and MEK-1 gene expression, and a significant reduction in autophagic machinery upon treatment.
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鬼针草和马齿苋是两种具有一系列生物活性类黄酮成分的杂草,因此它们可以作为潜在的健康补充剂和现成的天然抗氧化剂来源,以及药用应用中的有效成分。本研究旨在评估鬼针草和马齿苋提取物对 HepG2 细胞系的抗增殖活性。
方法:使用 MTT 法评估制备的提取物对 HepG2 细胞系和正常肝细胞的细胞毒性影响及其潜在 CC50。使用定量实时聚合酶链反应 (qRT-PCR) 检测处理细胞中 Raf-1、MEK-1、LC3B 和 Atg12 的相对基因表达,以检测细胞增殖因子和自噬相关基因的表达水平。还使用 ELISA 测定法对释放的白细胞介素-1β (IL-1β) 和白细胞介素-1α (IL-1α) 进行了定量分析。
结果:与正常细胞相比,鬼针草提取物对 HepG2 细胞系的活性表现出抗增殖作用。通过针对 Raf-1 和 MEK-1 的分子研究评估其细胞毒性,作为拟议的抗癌机制,并显示出对 Raf-1 和 MEK-1 基因表达的有希望的抑制活性。同样,在用鬼针草提取物预处理的 HepG2 细胞中,自噬相关基因 Atg12 和 LC3B 的减少进一步证实了其在诱导程序性细胞死亡 (PCD) 中的作用。ELISA 测定法显示,在用鬼针草提取物处理后,促炎细胞因子 IL-1α 和 IL-1β 的含量显著升高。
结论:本研究发现,鬼针草提取物在没有任何可检测的细胞毒性作用的情况下,对 Raf-1 和 MEK-1 基因表达具有潜在的抑制活性,并且在用鬼针草提取物处理后,自噬机制显著减少。