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

Fatemeh  Safari; Narjes  Rayat Azad; Ali  Alizadeh Ezdiny; Safoora  Pakizehkar; Zeinab  Khazaei Koohpar; Najmeh Ranji

doi:10.18502/ajmb.v14i1.8174

Fatemeh Safari Department of Biology, Faculty of Science, University of Guilan, Rasht, Iran
Narjes Rayat Azad Department of Biology, Faculty of Science, University of Guilan, Rasht, Iran
Ali Alizadeh Ezdiny Department of Biology, Faculty of Basic Sciences, Rasht Branch, Islamic Azad University, Rasht, Iran
Safoora Pakizehkar Cellular and Molecular Endocrine Research Center (CMERC), Research Institute for Endocrine Science, Shahid Beheshti University of Medical Sciences, Tehran, Iran
Zeinab Khazaei Koohpar Department of Cell and Molecular Biology, Faculty of Biological Sciences, Tonekabon Branch, Islamic Azad University, Tonekabon, Iran
Najmeh Ranji Department of Biology, Faculty of Basic Sciences, Rasht Branch, Islamic Azad University, Rasht, Iran

DOI: https://doi.org/10.18502/ajmb.v14i1.8174

Keywords: Apoptosis, Cell culture techniques, Green tea, Proto-oncogene proteins c-bcl-2, Up-regulation

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 Camellia sinensis) 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.