Role of bta‐miR‐204 in the regulation of adipocyte proliferation, differentiation, and apoptosis

Abstract

Adipocyte growth and development are complex and precisely orchestrated processes. Several microRNAs have been identified as critical regulators of the adipocyte growth and development. Recently, bta‐miR‐204 was found to be involved in adipogenesis; however, the underlying molecular mechanism involved in bta‐miR‐204‐mediated regulation of proliferation, differentiation, and apoptosis of adipocytes is not fully understood or elucidated. In this study, quantitative real‐time polymerase chain reaction (qRT‐PCR), Cell Counting Kit‐8, EdU, flow cytometer, Oil Red O staining, and the western blot assays were used to assess the role of bta‐miR‐204 in adipocyte growth and development. Overexpression of bta‐miR‐204 had no significant effect on 3T3‐L1 cell proliferation. The forced expression of bta‐miR‐204 promoted 3T3‐L1 cell differentiation. Meanwhile, overexpression of bta‐miR‐204 upregulated the expression of Bax and downregulated the expression of Bcl‐2 both at messenger RNA and protein levels, which suggested that bta‐miR‐204 can promote 3T3‐L1 cell apoptosis. Using bioinformatic analysis, dual‐luciferase reporter system and qRT‐PCR, TGFBR2, and ELOVL6 were identified as the direct target genes of bta‐miR‐204. Therefore, our study provides a novel insight into the role of bta‐miR‐204 in the regulation of adipocyte growth and development, which may provide a novel therapeutic alternative against obesity.

Mir-193b Regulates the Differentiation, Proliferation, and Apoptosis of Bovine Adipose Cells by Targeting the ACSS2/AKT Axis

“…RNA was reverse-transcribed using a reverse transcription kit (TaKaRa Biotech Co. Ltd.), in accordance with the manufacturer’s instructions. Real-time quantitative PCR (RT-qPCR) was performed using the SYBR green detection dye and the CFX96 real-time qPCR detection system, in accordance with the protocol reported by Jin et al [ 9 ] The expression of bta-miR-193b was normalized to that of U6. The expression of ACSS2 was normalized to that of GAPDH .…”

“…Finally, the oil red O dye was immediately discarded, and cells were washed four times with PBS. Images were captured under a microscope [ 9 ]. The oil red O was then eluted from the stained cells with isopropanol, and quantified using a microplate reader at 510 nm.…”

“…Most miRNAs play vital roles in regulating cell proliferation, differentiation, apoptosis, and other biological processes [ 7 ]. Recent studies have demonstrated that miRNAs play an important role in adipogenesis [ 8 , 9 ]. For example, miR-204 targets sirtuin 1 ( SIRT1 ) to promote adipogenesis, while miR-93 regulates obesity by inhibiting sirtuin 7 ( Sirt7 ) and T-box 3 (Tbx3) [ 10 ].…”

miR-205 Expression Elevated With EDS Treatment and Induced Leydig Cell Apoptosis by Targeting RAP2B via the PI3K/AKT Signaling Pathway

“…Cells were seeded in 6-well plates and treated for 48 h, then trypsinized and centrifuged at 800 g for 5 min, resuspended the cells with chilled DPBS, and slowly added an appropriate amount of 100% absolute ethanol to achieve a final concentration of 70%, stored at −20 • C. Subsequently, treated with the cell-cycle-staining kit (Multisciences, Hangzhou, China) and analyzed by flow cytometry (BD FACSAria III, BD Biosciences) (Jin et al, 2019).…”

circFLT1 and lncCCPG1 Sponges miR-93 to Regulate the Proliferation and Differentiation of Adipocytes by Promoting lncSLC30A9 Expression

“…In the field of non-coding RNA, microRNAs (miRNAs) are known to play important regulatory roles in adipogenesis. For example, miR-204 facilitates adipocyte differentiation by targeting Sirt1 , 5 , 6 and miR-93 can regulate adipocyte differentiation by inhibiting Sirtuin-7 and T-box3 . 7 …”