Gene expression profiling analysis of stress-sensitive genes and their potential functions in myoblasts

doi:10.19439/j.sjos.2025.01.002

Abstract

Abstract: PURPOSE: To screen the stress-sensitive genes in myoblasts and reveal the potential target genes and their regulatory mechanisms of facial muscle remodeling induced by functional orthopaedic force. METHODS: The procedure involved the use of gene microarray technology to identify the differentially expressed genes(DEGs) in myoblasts. DEGs were then categorized by Gene Ontology(GO) and Kyoto Encyclopedia of Genes and Genomes(KEGG) enrichment analyses. Furthermore, real-time quantitative PCR(qRT-PCR) was used to verify the DEGs. Western blot, transmission electron microscopy(TEM), and confocal laser scanning microscope(CLSM) were employed to detect the effect of stress on autophagy in myoblasts. The data were analyzed by SPSS 17.0 software package. RESULTS: A total of 1 410 DEGs were identified in stretched myoblasts, with 788 up-regulated and 622 down-regulated genes. GO enrichment analysis indicated that DEGs were primarily involved in signal transduction, biopolymer metabolic process, and protein metabolic process. KEGG analysis revealed that DEGs were primarily associated with ECM-receptor interaction, pathway in cancer, MAPK signaling pathway, focal adhesion and lysosome. Both TEM and CLSM showed that stress could promote the formation of autophagosomes, and Western blot demonstrated that stress could promote the expression of autophagy-related molecules Beclin-1 and LC3-II. Rapamycin could enhance all the above processes, while 3-MA could inhibit them. CONCLUSIONS: Autophagy may play an important role in the regulation of myoblast fate induced by cyclic tensile stress.

Key words: Cyclic stretch, Myoblast, Gene microarray, Signal pathway, Autophagy

CLC Number:

R783

ZHENG Yao, YU Shao-yang, YAN Xiao, LI Jian-ping, ZHANG Qiang, YUAN Xiao. Gene expression profiling analysis of stress-sensitive genes and their potential functions in myoblasts[J]. Shanghai Journal of Stomatology, 2025, 34(1): 7-13.

References

[1] Chen B, You W, Wang Y, et al.The regulatory role of Myomaker and Myomixer-Myomerger-Minion in muscle development and regeneration[J]. Cell Mol Life Sci, 2020, 77(8): 1551-1569.
[2] Fu S, Lin X, Yin L, et al.Androgen receptor regulates the proliferation of myoblasts under appropriate or excessive stretch through IGF-1 receptor mediated p38 and ERK1/2 pathways[J]. Nutr Metab (Lond), 2021, 18(1): 85.
[3] Zhang SJ, Truskey GA, Kraus WE.Effect of cyclic stretch on beta1D-integrin expression and activation of FAK and RhoA[J]. Am J Physiol Cell Physiol, 2007, 292(6): C2057-C2069.
[4] Du Y, Yang F, Lv D, et al.MiR-147 inhibits cyclic mechanical stretch-induced apoptosis in L6 myoblasts via ameliorating endoplasmic reticulum stress by targeting BRMS1[J]. Cell Stress Chaperones, 2019, 24(6): 1151-1161.
[5] Haga JH, Li YS, Chien S.Molecular basis of the effects of mechanical stretch on vascular smooth muscle cells[J]. J Biomech, 2007, 40(5): 947-960.
[6] Li M, Han Y, Wang S, et al.Becker muscular dystrophy: case report, review of the literature, and analysis of differentially expressed hub genes[J]. Neurol Sci, 2022, 43(1): 243-253.
[7] Palmieri O, Mazza T, Merla A, et al.Gene expression of muscular and neuronal pathways is cooperatively dysregulated in patients with idiopathic achalasia[J]. Sci Rep, 2016, 6: 31549.
[8] Bonafiglia JT, Menzies KJ, Gurd BJ.Gene expression variability in human skeletal muscle transcriptome responses to acute resistance exercise[J]. Exp Physiol, 2019, 104(5): 625-629.
[9] Zhang Q, Liu J, Chen S, et al.Caspase-12 is involved in stretch-induced apoptosis mediated endoplasmic reticulum stress[J]. Apoptosis, 2016, 21(4): 432-442.
[10] Sicari F, Merlo EM, Gentile G, et al.Body image and psychological impact of dental appearance in adolescents with malocclusion: a preliminary exploratory study[J]. Children (Basel), 2023, 10(10):1619.
[11] Volpato GH, de Almeida-Pedrin RR, Oltramari PVP, et al. Self-perception of facial esthetics by patients with different profiles compared with assessments of orthodontists and lay people[J]. Am J Orthod Dentofacial Orthop, 2020, 158(6): 840-848.
[12] Liu S, Gao F, Wen L, et al.Osteocalcin induces proliferation via positive activation of the PI3K/Akt, P38 MAPK pathways and promotes differentiation through activation of the GPRC6A-ERK1/2 pathway in C2C12 myoblast cells[J]. Cell Physiol Biochem, 2017, 43(3): 1100-1112.
[13] Chellini F, Tani A, Parigi M, et al.HIF-1α/MMP-9 axis is required in the early phases of skeletal myoblast differentiation under normoxia condition in vitro[J]. Cells, 2023, 12(24): 2851.
[14] Song Jt, Hu B, Qu Hy, et al.Mechanical stretch modulates microRNA 21 expression, participating in proliferation and apoptosis in cultured human aortic smooth muscle cells[J]. PLoS One, 2012, 7(10): e47657.
[15] Yang D, Wei GY, Li M, et al.Cyclic tensile strain facilitates proliferation and migration of human aortic smooth muscle cells and reduces their apoptosis via miRNA-187-3p[J]. Bioengineered, 2021, 12(2): 11439-11450.
[16] Hyldahl RD, Nelson B, Xin L, et al.Extracellular matrix remodeling and its contribution to protective adaptation following lengthening contractions in human muscle[J]. FASEB J, 2015, 29(7): 2894-2904.
[17] Natera-de Benito D, Jurgens JA, Yeung A, et al. Recessive variants in COL25A1 gene as novel cause of arthrogryposis multiplex congenita with ocular congenital cranial dysinnervation disorder[J]. Hum Mutat, 2022, 43(4): 487-498.
[18] Guo Y, Zhang CQ, Zeng QC, et al.Mechanical strain promotes osteoblast ECM formation and improves its osteoinductive potential[J]. Biomed Eng Online, 2012, 11: 80.
[19] Tipbunjong C, Khuituan P, Kitiyanant Y, et al.Diarylheptanoid 1-(4-hydroxyphenyl)-7-phenyl-(6E)-6-hepten-3-one enhances C2C12 myoblast differentiation by targeting membrane estrogen receptors and activates Akt-mTOR and p38 MAPK-NF-κB signaling axes[J]. J Nat Med, 2019, 73(4): 735-744.
[20] Andersen JI, Pennisi CP, Fink T, et al.Focal adhesion kinase activation is necessary for stretch-induced alignment and enhanced differentiation of myogenic precursor cells[J]. Tissue Eng Part A, 2018, 24(7-8): 631-640.
[21] Wang F, Wei ZL, Sun XR, et al.Apoptosis inducing factor is involved in stretch-induced apoptosis of myoblast via a caspase-9 independent pathway[J]. J Cell Biochem, 2017, 118(4): 829-838.
[22] Tan J, Kuang W, Jin Z, et al.Inhibition of NFkappaB by activated c-Jun NH2 terminal kinase 1 acts as a switch for C2C12 cell death under excessive stretch[J]. Apoptosis, 2009, 14(6): 764-770.
[23] Debnath J, Gammoh N, Ryan KM.Autophagy and autophagy-related pathways in cancer[J]. Nat Rev Mol Cell Biol, 2023, 24(8): 560-575.
[24] Lorzadeh S, Kohan L, Ghavami S, et al.Autophagy and the Wnt signaling pathway: a focus on Wnt/β-catenin signaling[J]. Biochim Biophys Acta Mol Cell Res, 2021, 1868(3): 118926.
[25] Denton D, Xu T, Kumar S.Autophagy as a pro-death pathway[J]. Immunol Cell Biol, 2015, 93(1): 35-42.