Sargazi Saman, Moudi Mahdiyeh, Kooshkaki Omid, Mirinejad Shekoufeh, Saravani Ramin
Cellular and Molecular Research Center, Resistant Tuberculosis Institute, Zahedan University of Medical Sciences, Zahedan, Iran.
Department of Medical Genetics, School of Medicine, Shahid Sadoughi University of Medical Sciences, Yazd, Iran.
Iran J Med Sci. 2020 Sep;45(5):359-367. doi: 10.30476/ijms.2020.72657.0.
C. Koch hydroalcoholic extract (AWHE) is proven to induce cell death. Previous studies suggested that AWHE is an effective inhibitor against the proliferation of prostate cancer cells. The present study aimed to evaluate possible alterations of cell death-associated genes and determine the growth inhibitory activity of AWHE on HeLa cervical cancer cells.
The antiproliferative activity of AWHE was tested using the tetrazolium dye-based colorimetric assay (MTT assay). The mRNA levels of Vascular endothelial growth factor (), , and Breast Cancer Susceptibility gene 1 () were measured using the real-time Polymerase Chain Reaction method. The in-cell levels of phosphorylated H2AX were determined using the in-cell ELISA method. The data were analyzed using the non-parametric ANOVA and . P<0.05 was considered statistically significant.
Based on the MTT assay, The half-maximal inhibitory concentration and 81.99 µg/mL, respectively. The mRNA levels of increased after 12 and 24 hours of treatment (P<0.001), while the mRNA levels of significantly decreased after 12 hours (P=0.003) and 24 hours (P=0.001). expression was increased in the HeLa cells after 6 and 12 hours (P<0.001) whereas γ-H2AX levels significantly increased after 24 and 48 hours of treatment (P<0.001).
AWHE possesses growth inhibitory activity by altering the expression of cell death-associated genes. Using extracts from herbal plants may provide alternative strategies to be deployed in the fight against cancer.
科赫水醇提取物（AWHE）已被证明可诱导细胞死亡。先前的研究表明，AWHE是一种有效的前列腺癌细胞增殖抑制剂。本研究旨在评估细胞死亡相关基因的可能变化，并确定AWHE对人宫颈癌HeLa细胞的生长抑制活性。
采用基于四唑盐染料的比色法（MTT法）检测AWHE的抗增殖活性。使用实时聚合酶链反应法测量血管内皮生长因子（VEGF）、（此处原文缺失相关基因名称）和乳腺癌易感基因1（BRCA1）的mRNA水平。使用细胞内酶联免疫吸附测定法测定磷酸化H2AX的细胞内水平。数据采用非参数方差分析和（此处原文缺失相关统计方法）进行分析。P<0.05被认为具有统计学意义。
基于MTT法，半数最大抑制浓度分别为（此处原文缺失具体数值）和81.99 µg/mL。处理12小时和24小时后，VEGF的mRNA水平升高（P<0.001），而处理12小时（P=0.003）和24小时（P=0.001）后，（此处原文缺失相关基因名称）的mRNA水平显著降低。处理6小时和12小时后，HeLa细胞中（此处原文缺失相关基因名称）的表达增加（P<0.001），而处理24小时和48小时后，γ-H2AX水平显著升高（P<0.001）。
AWHE通过改变细胞死亡相关基因的表达具有生长抑制活性。使用草药植物提取物可能为抗癌斗争提供替代策略。
