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利用3D细胞培养模型上调LNCaP细胞中miR-181a表达来研究绿茶的抗肿瘤活性

Antitumor Activities of Green Tea by Up-regulation of miR-181a Expression in LNCaP Cells Using 3D Cell Culture Model.

作者信息

Safari Fatemeh, Rayat Azad Narjes, Alizadeh Ezdiny Ali, Pakizehkar Safura, Khazaei Koohpar Zeinab, Ranji Najmeh

机构信息

Department of Biology, Faculty of Science, University of Guilan, Rasht, Iran.

Department of Biology, Faculty of Basic Sciences, Rasht Branch, Islamic Azad University, Rasht, Iran.

出版信息

Avicenna J Med Biotechnol. 2022 Jan-Mar;14(1):89-94. doi: 10.18502/ajmb.v14i1.8174.

Abstract

BACKGROUND

Prostate Cancer (PCa) is the major reason for the high mortality rates among men worldwide. In fact, current therapeutic approaches are not successful. It appears that discovering more effective methods considering several parameters such as availability, low cost, and no toxicity to normal cells is one of the biggest challenges for interested researchers. Green tea (extracted from the plant ) with high level of polyphenolic compounds and as the most globally consumed beverage has attracted considerable interest. MicroRNAs (or miRNAs) were considered as novel tools in cancer therapy which modulate various biological events in cell by regulation of gene expression. The aim of the current study was to evaluate the antitumor activity of green tea in LNCaP cells through up-regulation of miR-181a expression.

METHODS

First, LNCaP cells were cultured and by using quantitative real time PCR (qRT-PCR) and western blot methods, the expression levels of Bax and BCL2 were analyzed. Next, a 3D cell culture model was applied to evaluate the expression of miRNA-181a in LNCaP cells.

RESULTS

It was shown that green tea induced cellular apoptosis. The high number of apoptotic nuclei was also shown by using DAPI staining. The inhibition of tumor growth was revealed by analyzing the size and number of spheroids. Also, up-regulation of miR-181a expression in LNCaP cells was revealed after treatment with green tea.

CONCLUSION

Our results are helpful to design antitumor regimens based on consumption of green tea through up-regulation of miRNA-181a expression and induction of apoptosis.

摘要

背景

前列腺癌(PCa)是全球男性高死亡率的主要原因。事实上,目前的治疗方法并不成功。考虑到可用性、低成本以及对正常细胞无毒性等多个参数,发现更有效的方法似乎是相关研究人员面临的最大挑战之一。绿茶(从植物中提取)富含多酚类化合物,是全球消费最多的饮品,已引起了相当大的关注。微小RNA(或miRNA)被认为是癌症治疗中的新型工具,可通过调节基因表达来调控细胞中的各种生物学事件。本研究的目的是通过上调miR-181a的表达来评估绿茶对LNCaP细胞的抗肿瘤活性。

方法

首先,培养LNCaP细胞,并使用定量实时PCR(qRT-PCR)和蛋白质印迹法分析Bax和BCL2的表达水平。接下来,应用三维细胞培养模型评估LNCaP细胞中miRNA-181a的表达。

结果

结果表明绿茶可诱导细胞凋亡。通过DAPI染色也显示出大量凋亡细胞核。通过分析球体的大小和数量揭示了肿瘤生长的抑制。此外,用绿茶处理后,LNCaP细胞中miR-181a的表达上调。

结论

我们的结果有助于通过上调miRNA-181a的表达和诱导凋亡来设计基于饮用绿茶的抗肿瘤方案。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4678/9017474/671a5792fc50/AJMB-14-89-g001.jpg

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