miR-149-5p Regulates Goat Hair Follicle Stem Cell Proliferation and Apoptosis by Targeting the CMTM3/AR Axis During Superior-Quality Brush Hair Formation

Abstract

The Yangtze River Delta white goat is a unique goat species that can produce superior quality brush hair. CKLF-like MARVEL transmembrane domain-containing 3 (CMTM3), which influences the transcriptional activity of androgen receptor (AR), was identified as a candidate gene related to superior-quality brush hair formation. CMTM3 is generally expressed at low levels, but miR-149-5p is highly expressed in the skin tissues of these goats. The mechanism by which CMTM3 regulates the proliferation and apoptosis of goat hair follicle stem cells has not been elucidated. Here, RT-qPCR, western blotting, 5-ethynyl-2′-deoxyuridine (EdU), cell cycle, apoptosis, and dual-luciferase assays were used to investigate the role and regulatory mechanism of CMTM3 and miR-149-5p. Functional studies showed that CMTM3 overexpression inhibited proliferation and induced apoptosis in cultured hair follicle stem cells, whereas silencing CMTM3 markedly facilitated cell proliferation and deterred apoptosis in cultured hair follicle stem cells. Then, using bioinformatic predictions and the aforementioned assays, including dual-luciferase assays, RT-qPCR, and western blotting, we confirmed that miR-149-5p targets CMTM3 and preliminarily investigated the interaction between CMTM3 and AR in goat hair follicle stem cells. Furthermore, miR-149-5p overexpression significantly accelerated the proliferation and attenuated the apoptosis of hair follicle stem cells. Conversely, miR-149-5p inhibition suppressed the proliferation and induced the apoptosis of hair follicle stem cells. These results reveal a miR-149-5p-related regulatory framework for the miR-149-5p/CMTM3/AR axis during superior quality brush hair formation, in which CMTM3 plays a negative role.

The Circular RNA CircCOL1A1 Functions as a miR-149-5p Sponge to Regulate the Formation of Superior-Quality Brush Hair via the CMTM3/AR Axis

“…RegRNA 2.0 (http://regrna2.mbc.nctu.edu.tw/index.html) and RNAhybrid (v2.1.2) software were used to obtain the secondary structure and information on the binding sites between circCOL1A1 and miR-149-5p. The miR-149-5p/ CMTM3/AR axis was constructed and identified in our previous study (Wang et al, 2020). Sanger sequencing, PCR with divergent and convergent primers, RNase R treatment, actinomycin D treatment, and cytoplasmic and nuclear RNA purification assays were used to identify the structure and specificity of circCOL1A1.…”

“…4) CMTM3 was confirmed as a target gene of miR-149-5p, and during superiorquality brush hair formation, miR-149-5p could regulate hair follicle stem cell proliferation and apoptosis by targeting the CMTM3/AR axis (Wang et al, 2020). 5) CMTM3 knockdown accelerates hair follicle stem cell proliferation and promotes AR expression, while overexpression of CMTM3 significantly inhibits the proliferation of stem cells and is accompanied by a reduction in the AR expression level (Wang et al, 2020;Wang et al, 2021). This critical evidence suggests that a series of protein-coding genes and noncoding RNAs (ncRNAs) are important for the formation of superior-quality brush hair.…”

“…3) A high methylation level of CMTM3 could promote the formation of superior-quality brush hair, and CMTM3, also known as CKLFSF3, could modulate the transcription level of androgen receptor (AR) (Zhong et al, 2006;Wang et al, 2018). 4) CMTM3 was confirmed as a target gene of miR-149-5p, and during superiorquality brush hair formation, miR-149-5p could regulate hair follicle stem cell proliferation and apoptosis by targeting the CMTM3/AR axis (Wang et al, 2020). 5) CMTM3 knockdown accelerates hair follicle stem cell proliferation and promotes AR expression, while overexpression of CMTM3 significantly inhibits the proliferation of stem cells and is accompanied by a reduction in the AR expression level (Wang et al, 2020;Wang et al, 2021).…”

A systematic summary of survival and death signalling during the life of hair follicle stem cells

“…Most important, caspases and Bcl-2 may be targets that regulate the apoptosis for HFSCs. For example, the Jian group revealed that a miR-149-5p inhibitor can suppress the proliferation and trigger the apoptosis of HFSCs, while miR-149-5p can upregulate the expression of Bcl-2 and downregulate caspase 3, as well as induce anti-apoptotic responses during superior-quality brush hair formation [ 170 ].…”

MiR-149-5p: An Important miRNA Regulated by Competing Endogenous RNAs in Diverse Human Cancers

“…Functional studies indicated miR-149-5p plays an important role in the formation of high-quality hair traits. Through the post-transcriptional mechanism, it can inhibit the expression of CKLF-like MARVEL transmembrane domain-containing 3 (CMTM3), promote the proliferation of goat hair follicle stem cells and inhibit the apoptosis of hair follicle stem cells (33).…”