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绿色合成的银纳米粒子通过激活 MLH1 基因表达诱导 CACO2 癌细胞凋亡。

Green-synthesized silver nanoparticles induced apoptotic cell death in CACO2 cancer cells by activating MLH1 gene expression.

机构信息

Agriculture Biotechnology Research Institute, University of Zabol, Zabol, Iran.

Department of Agronomy and Plant Breeding, Agriculture Institute, Research Institute of Zabol, Zabol, Iran.

出版信息

Sci Rep. 2024 Nov 28;14(1):29601. doi: 10.1038/s41598-024-80809-0.

Abstract

MLH1 (Mult homolog1) gene is the main element of Multlα heterodimer and plays a role in the repair of base-base mismatches and deletion and addition loops. When the MLH1 protein is not present, the number of errors that remain unrepaired increases, and this can lead to the formation of tumors in the body. Colorectal cancer is the third most common type of cancer in terms of incidence and the third type of cancer in terms of mortality worldwide. In this regard, the present study was conducted with the aim of investigating the effect of biological silver nanoparticles with anticancer properties and MLH1 gene expression on cancer cell samples of people with colorectal cancer. In this study, the green synthesis of silver nanoparticles was carried out by precipitation method with reduction of silver ions by leaf and flower extract of Moringa oleifera plant. Then, silver nanoparticles were confirmed using UV-visible spectroscopy, transmission and scanning electron microscopy. The cytotoxic effects of nanoparticles on cells were evaluated by MTT colorimetric method within 42 h. After RNA extraction of treated cells, cDNA synthesis and primers were designed by the Exon-Exon Junction method and the Real-time Polymerase test for MLH1 and Beta-actin genes was repeated three times. The final analysis of the results was done using Graphpad Prism and Rest 2009 software. The presence of a peak at the wavelength of 234 nm for the synthesized silver nanoparticles was confirmed by ultraviolet-visible spectroscopic analysis. The morphological study on the size and shape of the silver nanoparticles showed that the nanoparticles are spherical and have a size between 40 and 34 nanometers in diameter. The leaf extract typically produces smaller, more uniform particles than the flower extract. The Green-Synthesized Silver Nanoparticles of leaf extract were used to evaluation of induced Apoptotic Cell Death in CACO2 Cancer Cells by Activating MLH1 Gene Expression. MTT results showed that the anti-proliferative effect of nanoparticles depends on the concentration of synthesized silver nanoparticles. The treatment of MLH1 cancer cell line and normal with synthesized nanoparticles at a concentration of 0.44 micrograms per ml for 42 h showed the effects of cell toxicity. The percentage of cancer cell death under the influence of quercetin present in Moringa green bio-particles depends on the concentration and time, and this difference is statistically significant compared to the control group (P < 0.05). So that the lethal dose of the extract for 50% survival IC50 in a period of 48 h for intestinal cancer cells was equal to 21 μg/ml, and the expression ratio of the MLH1 gene in the tumor tissue to the adjacent healthy tissue has increased (P ≤ 0.05).

摘要

MLH1(多聚体同源物 1)基因是 Multlα 异二聚体的主要成分,在碱基对碱基错配和缺失及添加环的修复中发挥作用。当 MLH1 蛋白不存在时,未修复的错误数量增加,这可能导致体内肿瘤的形成。结直肠癌是全球发病率第三高的癌症,也是死亡率第三高的癌症。有鉴于此,本研究旨在探讨具有抗癌特性的生物银纳米粒子和 MLH1 基因表达对结直肠癌患者癌细胞样本的影响。在这项研究中,通过沉淀法用辣木叶和花提取物还原银离子,进行银纳米粒子的绿色合成。然后,通过紫外可见光谱、透射和扫描电子显微镜对银纳米粒子进行了确认。通过 MTT 比色法在 42 小时内评估纳米粒子对细胞的细胞毒性作用。用细胞提取 RNA 后,通过外显子-外显子连接法设计 cDNA 合成和引物,用实时聚合酶对 MLH1 和 Beta-actin 基因进行三次重复测试。使用 Graphpad Prism 和 Rest 2009 软件对结果进行最终分析。通过紫外可见光谱分析,确认合成银纳米粒子的波长为 234nm 处有峰存在。银纳米粒子的大小和形状的形态研究表明,纳米粒子是球形的,直径在 40 至 34 纳米之间。与花提取物相比,叶提取物通常产生更小、更均匀的颗粒。用叶提取物的绿色合成银纳米粒子来评价通过激活 MLH1 基因表达诱导 CACO2 癌细胞凋亡。MTT 结果表明,纳米粒子的抗增殖作用取决于合成银纳米粒子的浓度。在 42 小时内,用浓度为 0.44 微克/毫升的合成纳米粒子处理 MLH1 癌细胞系和正常细胞,显示出细胞毒性作用。在莫桑比克绿色生物颗粒中存在的槲皮素的影响下,癌细胞的死亡率百分比取决于浓度和时间,与对照组相比,这一差异具有统计学意义(P<0.05)。因此,该提取物对肠道癌细胞的 50%存活 IC50 的致死剂量在 48 小时内等于 21μg/ml,肿瘤组织中 MLH1 基因的表达比值与相邻健康组织相比增加(P≤0.05)。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ca7b/11604737/936bdd91310d/41598_2024_80809_Fig1_HTML.jpg

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