chi-miR-487b-3p Inhibits Goat Myoblast Proliferation and Differentiation by Targeting IRS1 through the IRS1/PI3K/Akt Signaling Pathway

Lyu, Ming; Wang, Xu; Meng, Xiangyu; Qian, Hongrun; Li, Qian; Ma, Ben; Zhang, Zhiying; Xu, Kun

doi:10.3390/ijms23010115

Cited by 15 publications

(15 citation statements)

References 42 publications

(54 reference statements)

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“…Meanwhile, si-circ_003628 increased the expression level of p27 . It has been reported that Pax7 , CDK2 , CDK4 and CyclinD1 in expression promoted the proliferation of goat SMSCs, while p27 inhibited the proliferation of goat SMSCs [ 27 , 46 ]. This taken together with the inhibited effect of si-circ_003628 on goat SMSCs, suggests that circ_003628 promotes proliferation of goat SMSCs by regulating the expression of the proliferation marker genes, eventually leading to the hypertrophy of muscle fibers and increase of skeletal muscle mass in goats.…”

Section: Discussionmentioning

confidence: 99%

Characteristics and Expression of circ_003628 and Its Promoted Effect on Proliferation and Differentiation of Skeletal Muscle Satellite Cells in Goats

Zhen

Wang

Luo

et al. 2022

Animals

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In our previous a study, circ_003628 was one of the most highly expressed circular RNAs (circRNAs) in the Longissimus dorsi muscle of goats found by RNA-seq, suggesting that the circRNA may be important for caprine muscle growth and development. However, there have been no reports describing the molecular mechanisms by which circ_003628 regulates the activities of goat skeletal muscle satellite cells (SMSCs). In this study, reverse transcriptase-PCR (RT-PCR) and DNA sequencing were used to validate the authenticity of circ_003628, and its characteristics, expression profile and effect on goat SMSCs were also studied using real-time quantitative-PCR (RT-qPCR), EdU, CCK-8 and immunofluorescence assays. Circ_003628 is partially originated from 13 exons, 12 introns and 3’-untranslated regions (UTR) of caprine Myosin Heavy Chain 1 (MYH1), and 25 exons and 5’ UTR of Myosin Heavy Chain 4 (MYH4), as well as intergenic sequences between the two genes. A total of 77.07% of circ_003628 were located in the nuclei of goat SMSCs, while 22.93% were expressed in the cytoplasm. The circRNAs were only expressed in triceps brachii, quadriceps femoris and longissimus dorsi muscle tissues in nine caprine tissues investigated, with the highest expression level in longissimus dorsi muscle. The expression level of circ_003628 gradually increased during differentiation periods of goat SMSCs and reached the maximum on day 6 after differentiation. The small interfering RNA of circ_003628 (named si-circ_003628) inhibited the viability and proliferation of goat SMSCs, and also decreased the expression of four cell proliferation marker genes: paired box 7 (Pax7), cyclin-dependent kinase 2 (CDK2), CDK4 and CyclinD1 in goat SMSCs. Transfection of si-circ_003628 significantly decreased the area of MyHC-labeled myotubes of goat SMSCs, as well as the expression levels of three differentiation marker genes: myosin heavy chain (MyHC), myogenin (MyoG), and myocyte enhancer factor 2C (MEF2C). These results suggest that circ_003628 promotes the viability, proliferation, and differentiation of goat SMSCs, and they also provide an improved understanding of the roles of circ_003628 in skeletal muscle growth and development in goats.

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Section: Discussionmentioning

confidence: 99%

Characteristics and Expression of circ_003628 and Its Promoted Effect on Proliferation and Differentiation of Skeletal Muscle Satellite Cells in Goats

Zhen

Wang

Luo

et al. 2022

Animals

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“…During reproduction, regulating cell cycle-related factors contributes to better follicle development and protects female fertility ( 30 , 31 ). Activation of the PI3K-Akt signaling pathway and Oocyte meiosis can promote oocyte maturation ( 32 – 34 ). Our results suggest that the cell cycle, PI3K-Akt signaling pathway, and Oocyte meiosis play important roles in follicle development.…”

Section: Discussionmentioning

confidence: 99%

Identification of hub genes associated with follicle development in multiple births sheep by WGCNA

2022

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Sheep exhibit a distinct estrous cycle that includes four different phases: proestrus, estrus, late estrus, and luteal phase. As the estrous cycle repeats, follicular development regularly alternates. We thus investigated ovarian transcriptome data from each of the four phases using weighted gene co-expression network analysis (WGCNA) to identify modules, pathways, and genes essential to follicle growth and development. We clustered mRNA and long non-coding RNA (lncRNA) into different modules by WGCNA, and calculated correlation coefficients between genes and Stages of the estrous cycle. Co-expression of the black module (cor = 0.81, P<0.001) and the yellow module (cor = 0.61, P<0.04) was found to be critical for follicle growth and development. A total of 2066 genes comprising the black and yellow modules was used for functional enrichment. The results reveal that these genes are mainly enriched in Cell cycle, PI3K-Akt signaling pathway, Oocyte meiosis, Apoptosis, and other important signaling pathways. We also identified seven hub genes (BUB1B, MAD2L1, ASPM, HSD3B1, WDHD1, CENPA, and MXI1) that may play a role in follicle development. Our study may provide several important new markers allowing in depth exploration of the genetic basis for multiparous reproduction in sheep.

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“…Glyceraldehyde 3-phosphate dehydrogenase was used as an endogenous control gene for mRNA, and U6 for miRNA. Relative quantitative levels were calculated based on the 2 −ΔΔCt method [ 4 ].…”

Section: Methodsmentioning

confidence: 99%

“…The formation of animal muscle tissue mainly depends upon the accumulation of proteins and the coordination of muscle cell proliferation, differentiation, and apoptosis, and includes three main stages: myotube formation, myotube differentiation into muscle fibers, and muscle fiber growth and maturation [ 1 ]. Skeletal muscle satellite cells (SMSCs) have stem-cell-like properties and are involved in postnatal skeletal muscle regeneration and growth [ 2 , 3 , 4 ]. Due to their myogenic and adipogenic differentiation capacity, SMSCs have become a suitable model for investigating postnatal muscle growth and intramuscular adipogenesis [ 5 ].…”

Section: Introductionmentioning

confidence: 99%

“…They are highly evolutionarily conserved and play important roles in the complex and dynamic network that regulates development. miR-487b-3p downregulated IRS1 expression to suppress goat myoblast cell proliferation and differentiation through the IRS1/PI3K/Akt pathway [ 4 ], miR-181a could suppress SMSC proliferation [ 2 ], and miR-223-3p was involved in muscle regeneration via the regulation of inflammatory cytokine secretion in the skeletal muscle microenvironment [ 7 ].…”

Section: Introductionmentioning

confidence: 99%

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chi-miR-99b-3p Regulates the Proliferation of Goat Skeletal Muscle Satellite Cells In Vitro by Targeting Caspase-3 and NCOR1

Liao

Dai

et al. 2022

Animals

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We previously found that chi-miR-99b-3p was highly expressed in the skeletal muscle of 7-month-old (rapid growth period) goats and speculated that it may be associated with muscle development. To further investigate the role of chi-miR-99b-3p in goats, we found that chi-miR-99b-3p acted as a myogenic miRNA in the regulation of skeletal muscle development. Dual-luciferase reporter assays, qRT-PCR, and Western blot results confirmed that Caspase-3 and nuclear receptor corepressor 1 were direct targets for chi-miR-99b-3p as their expression was inhibited by this miR. Cell proliferation and qRT-PCR assays showed that chi-miR-99b-3p promoted proliferation through relevant targets and intrinsic apoptosis-related genes in goat skeletal muscle satellite cells (SMSCs), whereas inhibition of chi-miR-99b-3p had the opposite effect. Furthermore, integrative transcriptomic analysis revealed that overexpression of chi-miR-99b-3p induced various differentially expressed (DE) genes mainly associated with the cell cycle, relaxin signaling pathway, DNA replication, and protein digestion and absorption. Notably, most of the cell-cycle-related genes were downregulated in SMSCs after miR-99b-3p upregulation, including the pro-apoptosis-related gene BCL2. In addition, 47 DE miRNAs (16 upregulated and 31 downregulated) were determined by Small RNA-sequencing in SMSCs after chi-miR-99b-3p overexpression. Based on the KEGG enrichment analysis, we found that these DE miRNAs were involved in the biological pathways associated with the DE genes. Our study demonstrated that chi-miR-99b-3p was an effective facilitator of goat SMSCs and provided new insights into the mechanisms by which miRNAs regulate skeletal muscle growth in goats.

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chi-miR-487b-3p Inhibits Goat Myoblast Proliferation and Differentiation by Targeting IRS1 through the IRS1/PI3K/Akt Signaling Pathway

Cited by 15 publications

References 42 publications

Characteristics and Expression of circ_003628 and Its Promoted Effect on Proliferation and Differentiation of Skeletal Muscle Satellite Cells in Goats

Characteristics and Expression of circ_003628 and Its Promoted Effect on Proliferation and Differentiation of Skeletal Muscle Satellite Cells in Goats

Identification of hub genes associated with follicle development in multiple births sheep by WGCNA

chi-miR-99b-3p Regulates the Proliferation of Goat Skeletal Muscle Satellite Cells In Vitro by Targeting Caspase-3 and NCOR1

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