LiCl与MSAB对C<sub>2</sub>C<sub>12</sub>细胞Wnt/β-catenin信号通路及增殖分化的影响

doi:10.7668/hbnxb.20195994

摘要/Abstract

摘要：

探讨Wnt/β-catenin信号通路激活剂(LiCl)与抑制剂(MSAB)对C₂C₁₂细胞中该信号通路的作用,以及该通路被激活或抑制后对C₂C₁₂细胞增殖和成肌分化的作用。以C₂C₁₂细胞作为模型,采用不同浓度LiCl(0,15,25 mmol/L)及IWR-1、MSAB(0,15,25 μmol/L)对细胞处理并诱导成肌分化,通过Western Blotting技术检测经典Wnt/β-catenin信号通路相关因子及细胞增殖和成肌分化标志基因蛋白的表达。结果表明,15 mmol/L LiCl对Wnt信号通路中β-catenin蛋白表达没有显著影响,而25 mmol/L LiCl能够促进C₂C₁₂细胞内β-catenin、Pax7、MyOG和MyHC的表达。IWR-1对β-catenin蛋白表达无显著影响,而15,25 μmol/L MSAB对β-catenin蛋白表达具有抑制作用,其中15 μmol/L MSAB的抑制效果尤为显著,明显降低了C₂C₁₂细胞内β-catenin、Pax7、MyOG和MyHC的表达水平。综上,25 mmol/L LiCl与15 μmol/L MSAB可分别有效激活和抑制C₂C₁₂细胞内Wnt信号通路,并对C₂C₁₂细胞增殖和成肌分化产生影响。

关键词: Wnt/β-catenin信号通路, LiCl, MSAB, C₂C₁₂细胞

Abstract:

The aim of this study was to investigate the effects of the activator (LiCl) and the inhibitor (MSAB) on the Wnt/β-catenin signaling pathway in C₂C₁₂ cells,as well as to explore how activation or inhibition of this pathway influences myogenic differentiation in C₂C₁₂ cells.This study utilized C₂C₁₂ cells as a model,the cells were treated with different concentrations of LiCl (0,15,and 25 mmol/L) as well as IWR-1 and MSAB (0,15,and 25 μmol/L) to induce differentiation.The expression of proteins related to the classical Wnt signaling pathway and myogenic differentiation cells was detected using Western Blotting technology.The results indicated that a concentration of 15 mmol/L LiCl did not had a significant effect on the β-catenin protein within the Wnt signaling pathway.In contrast,a concentration of 25 mmol/L LiCl promoted the expression of β-catenin,Pax7,MyOG,and MyHC in C₂C₁₂ cells.IWR-1 had no significant effect on β-catenin protein,whereas MSAB at concentrations of 15 and 25 μmol/L exhibited an inhibitory effect on the expression of β-catenin protein.Notably,the inhibition with 15 μmol/L MSAB was particularly pronounced,leading to a marked reduction in the expression levels of β-catenin,Pax7,MyOG,and MyHC in C₂C₁₂ cells.In conclusion,25 mmol/L LiCl and 15 μmol/L MSAB can effectively activate and inhibit the Wnt signaling pathway within C₂C₁₂ cells,thereby influencing myogenic differentiation in these cells.

Key words: Wnt/β-catenin signaling pathway, LiCl, MSAB, C₂C₁₂ cells

中图分类号:

S813.1遗传

韦海妹, 江美祺, 刘众志, 李新, 胡德宝, 郭益文, 丁向彬, 郭宏, 张林林. LiCl与MSAB对C₂C₁₂细胞Wnt/β-catenin信号通路及增殖分化的影响[J]. 华北农学报, 2026, 41(2): 232-238. doi: 10.7668/hbnxb.20195994.

WEI Haimei, JIANG Meiqi, LIU Zhongzhi, LI Xin, HU Debao, GUO Yiwen, DING Xiangbin, GUO Hong, ZHANG Linlin. The Effects of LiCl and MSAB on the Wnt/β-Catenin Signaling Pathway, Proliferation,and Differentiation in C₂C₁₂ Cells[J]. Acta Agriculturae Boreali-Sinica, 2026, 41(2): 232-238. doi: 10.7668/hbnxb.20195994.

http://www.hbnxb.net/CN/Y2026/V41/I2/232

图/表 6

图1 激活剂LiCl对β-catenin影响 A.Western Blotting 试验检测不同浓度LiCl对β-catenin蛋白的影响;B. A图中条带的量化图。相同字母表示差异不显著,不同字母表示差异显著(P<0.05)。图2—6同。

Fig.1 The impact of activator LiCl on β-catenin A. Detecting the effect of different concentrations of LiCl on β-catenin protein by Western Blotting;B. Quantitative graph of bands in Fig.A.The same letters indicate no significant difference, and different letters indicate a significant difference (P<0.05).The same as Fig.2—6.

图2 抑制剂IWR-1(A、B)与MSAB(C、D)对β-catenin影响

Fig.2 Effects of inhibitors IWR-1(A,B)and MSAB(C,D)on β-catenin

图3 LiCl激活Wnt/β-catenin并促进C₂C₁₂细胞增殖

Fig.3 The effect of LiCl activation on Wnt/β-catenin signaling and C₂C₁₂ cell proliferation

图4 LiCl激活Wnt/β-catenin信号通路并促进C₂C₁₂细胞成肌分化

Fig.4 The effect of LiCl on the activation of the Wnt/β-catenin signaling pathway and C₂C₁₂ cell differentiation

图5 MSAB对Wnt/β-catenin信号通路的抑制作用影响C₂C₁₂细胞的增殖

Fig.5 The inhibitory effect of MSAB on the Wnt/β-catenin signaling pathway influences the proliferation of C₂C₁₂ cells

图6 MSAB对C₂C₁₂细胞内Wnt/β-catenin信号通路及其分化的影响

Fig.6 The effect of MSAB on the Wnt/β-catenin signaling pathway and differentiation in C₂C₁₂ cells

参考文献 37

[1]	Jeong G J, Castels H, Kang I, Aliya B, Jang Y C. Nanomaterial for skeletal muscle regeneration[J]. Tissue Engineering and Regenerative Medicine, 2022, 19(2):253-261.doi:10.1007/s13770-022-00446-4.
[2]	Mo M J, Zhang Z H, Wang X T, Shen W J, Zhang L, Lin S D. Molecular mechanisms underlying the impact of muscle fiber types on meat quality in livestock and poultry[J]. Frontiers in Veterinary Science, 2023, 10:1284551.doi:10.3389/fvets.2023.1284551. URL
[3]	Vemula S, Muvavarirwa T, Doornbos F, Whitman M C. Neuromuscular junction development differs between extraocular and skeletal muscles and between different extraocular muscles[J]. Investigative Ophthalmology & Visual Science, 2024, 65(5):28.doi:10.1167/iovs.65.5.28.
[4]	Özen S D, Kir S. Ectodysplasin A2 receptor signaling in skeletal muscle pathophysiology[J]. Trends in Molecular Medicine, 2024, 30(5):471-483.doi:10.1016/j.molmed.2024.02.002. pmid: 38443222
[5]	Sousa-Victor P, García-Prat L, Muñoz-Cánoves P. Control of satellite cell function in muscle regeneration and its disruption in ageing[J]. Nature Reviews Molecular Cell Biology, 2022, 23(3):204-226.doi:10.1038/s41580-021-00421-2.
[6]	Schmidt M, Schüler S C, Hüttner S S, von Eyss B, von Maltzahn J. Adult stem cells at work:regenerating skeletal muscle[J]. Cellular and Molecular Life Sciences, 2019, 76(13):2559-2570.doi:10.1007/s00018-019-03093-6. pmid: 30976839
[7]	Buckingham M, Relaix F. PAX3 and PAX7 as upstream regulators of myogenesis[J]. Seminars in Cell & Developmental Biology, 2015, 44:115-125.doi:10.1016/j.semcdb.2015.09.017.
[8]	Yue Y Q, Wang Y H, Wen C L, Meng Y Y, Peng Y, Li X. Lnc-Malat1 promotes slow myofiber-type transformation through sponging miR-129-5p in C₂C₁₂ myotubes[J]. Experimental Cell Research, 2023, 431(1):113761.doi:10.1016/j.yexcr.2023.113761. URL
[9]	Zammit P S. Function of the myogenic regulatory factors Myf5,MyoD,Myogenin and MRF4 in skeletal muscle,satellite cells and regenerative myogenesis[J]. Seminars in Cell & Developmental Biology, 2017, 72:19-32.doi:10.1016/j.semcdb.2017.11.011.
[10]	朱春月, 杨培民, 刘忠航, 张伯序, 张健, 胡宗云. 刀鲚MyoG基因克隆和组织表达分析[J]. 天津农业科学, 2024, 30(10):33-39,49.doi:10.3969/j.issn.1006-6500.2024.10.006.
	Zhu C Y, Yang P M, Liu Z H, Zhang B X, Zhang J, Hu Z Y. Cloning and tissue expression analysis of MyoG gene in Coilia nasus[J]. Tianjin Agricultural Sciences, 2024, 30(10):33-39,49.
[11]	Agosti E, Antonietti S, Ius T, Fontanella M M, Zeppieri M, Panciani P P. Glioma stem cells as promoter of glioma progression:a systematic review of molecular pathways and targeted therapies[J]. International Journal of Molecular Sciences, 2024, 25(14):7979.doi:10.3390/ijms25147979. URL
[12]	Girardi F, Le Grand F. Wnt signaling in skeletal muscle development and regeneration[J]. Progress in Molecular Biology and Translational Science, 2018, 153:157-179.doi:10.1016/bs.pmbts.2017.11.026. pmid: 29389515
[13]	Liu D, Li S, Cui Y F, Tong H L, Li S F, Yan Y Q. Podocan affects C₂C₁₂ myogenic differentiation by enhancing Wnt/β-catenin signaling[J]. Journal of Cellular Physiology, 2019, 234(7):11130-11139.doi:10.1002/jcp.27763. URL
[14]	Adhikari A, Davie J. JARID2 and the PRC2 complex regulate skeletal muscle differentiation through regulation of canonical Wnt signaling[J]. Epigenetics & Chromatin, 2018, 11(1):46.doi:10.1186/s13072-018-0217-x.
[15]	Maeda K, Enomoto A, Hara A, Asai N Y, Kobayashi T, Horinouchi A, Maruyama S, Ishikawa Y, Nishiyama T, Kiyoi H, Kato T, Ando K, Weng L, Mii S, Asai M, Mizutani Y, Watanabe O, Hirooka Y, Goto H, Takahashi M. Identification of meflin as a potential marker for mesenchymal stromal cells[J]. Scientific Reports, 2016, 6:22288.doi:10.1038/srep22288. pmid: 26924503
[16]	Yui S, Azzolin L, Maimets M, Pedersen M T, Fordham R P, Hansen S L, Larsen H L, Guiu J, Alves M R P, Rundsten C F, Johansen J V, Li Y, Madsen C D, Nakamura T, Watanabe M, Nielsen O H, Schweiger P J, Piccolo S, Jensen K B. YAP/TAZ-dependent reprogramming of colonic epithelium links ECM remodeling to tissue regeneration[J]. Cell Stem Cell, 2018, 22(1):35-49.e7.doi:10.1016/j.stem.2017.11.001. pmid: 29249464
[17]	Zhang K, Zhang Y Y, Gu L J, Lan M M, Liu C C, Wang M, Su Y, Ge M X, Wang T T, Yu Y Y, Liu C, Li L, Li Q Y, Zhao Y F, Yu Z Q, Wang F D, Li N, Meng Q Y. Islr regulates canonical Wnt signaling-mediated skeletal muscle regeneration by stabilizing Dishevelled-2 and preventing autophagy[J]. Nature Communications, 2018, 9:5129.doi:10.1038/s41467-018-07638-4. pmid: 30510196
[18]	Kajiguchi T, Katsumi A, Tanizaki R, Kiyoi H, Naoe T. Y654 of β-catenin is essential for FLT3/ITD-related tyrosine phosphorylation and nuclear localization of β-catenin[J]. European Journal of Haematology, 2012, 88(4):314-320.doi:10.1111/j.1600-0609.2011.01738.x.
[19]	Yoshida C, Takeichi M. Teratocarcinoma cell adhesion:identification of a cell-surface protein involved in calcium-dependent cell aggregation[J]. Cell, 1982, 28(2):217-224.doi:10.1016/0092-8674(82)90339-7. pmid: 7060128
[20]	Finch P W, He X, Kelley M J, Üren A, Schaudies R P, Popescu N C, Rudikoff S, Aaronson S A, Varmus H E, Rubin J S. Purification and molecular cloning of a secreted,frizzled-related antagonist of Wnt action[J]. Proceedings of the National Academy of Sciences of the United States of America, 1997, 94(13):6770-6775.doi:10.1073/pnas.94.13.6770. pmid: 9192640
[21]	Zhao H, Ming T Q, Tang S, Ren S, Yang H, Liu M L, Tao Q, Xu H B. Wnt signaling in colorectal cancer:pathogenic role and therapeutic target[J]. Molecular Cancer, 2022, 21(1):144.doi:10.1186/s12943-022-01616-7. pmid: 35836256
[22]	Wang H Y, Zhang R, Wu X J, Chen Y F, Ji W, Wang J S, Zhang Y W, Xia Y, Tang Y Q, Yuan J X. The Wnt signaling pathway in diabetic nephropathy[J]. Frontiers in Cell and Developmental Biology, 2022, 9:701547.doi:10.3389/fcell.2021.701547. URL
[23]	Xue R Z, Lin W F, Sun J K, Watanabe M, Xu A B, Araki M, Nasu Y, Tang Z Y, Huang P. The role of Wnt signaling in male reproductive physiology and pathology[J]. Molecular Human Reproduction, 2021, 27(1):gaaa085.doi:10.1093/molehr/gaaa085.
[24]	Vuong L T, Mlodzik M. Different strategies by distinct Wnt-signaling pathways in activating a nuclear transcriptional response[J]. Current Topics in Developmental Biology, 2022, 149:59-89.doi:10.1016/bs.ctdb.2022.02.008. pmid: 35606062
[25]	Gajos-Michniewicz A, Czyz M. Wnt signaling in melanoma[J]. International Journal of Molecular Sciences, 2020, 21(14):4852.doi:10.3390/ijms21144852. URL
[26]	Neiheisel A, Kaur M, Ma N, Havard P, Shenoy A K. Wnt pathway modulators in cancer therapeutics:an update on completed and ongoing clinical trials[J]. International Journal of Cancer, 2022, 150(5):727-740.doi:10.1002/ijc.33811. URL
[27]	Xia M Y, Zhao X Y, Huang Q L, Sun H Y, Sun C, Yuan J, He C, Sun Y, Huang X, Kong W, Kong W J. Activation of Wnt/β-catenin signaling by lithium chloride attenuates d-galactose-induced neurodegeneration in the auditory cortex of a rat model of aging[J]. FEBS Open Bio, 2017, 7(6):759-776.doi:10.1002/2211-5463.12220. URL
[28]	Morgan J, Partridge T. Skeletal muscle in health and disease[J]. Disease Models & Mechanisms, 2020, 13(2):dmm042192.doi:10.1242/dmm.042192.
[29]	Hashemolhosseini S, Gessler L. Crosstalk among canonical Wnt and Hippo pathway members in skeletal muscle and at the neuromuscular junction[J]. Neural Regeneration Research, 2025, 20(9):2464-2479.doi:10.4103/nrr.nrr-d-24-00417. URL
[30]	Han S S, Cui C, He H R, Shen X X, Chen Y Q, Wang Y, Li D Y, Zhu Q, Yin H D. Myoferlin regulates Wnt/β-catenin signaling-mediated skeletal muscle development by stabilizing dishevelled-2 against autophagy[J]. International Journal of Molecular Sciences, 2019, 20(20):5130.doi:10.3390/ijms20205130. URL
[31]	Rudnicki M A, Williams B O. Wnt signaling in bone and muscle[J]. Bone, 2015, 80:60-66.doi:10.1016/j.bone.2015.02.009. pmid: 26453496
[32]	Patergnani S, Buchsbaum D J, Piazza G A. Editorial:targeting the Wnt/β-catenin signaling pathway in cancer[J]. Frontiers in Oncology, 2022, 12:1022174.doi:10.3389/fonc.2022.1022174. URL
[33]	Damsky W E, Curley D P, Santhanakrishnan M, Rosenbaum L E, Platt J T, Gould Rothberg B E, Taketo M M, Dankort D, Rimm D L, McMahon M, Bosenberg M. β-catenin signaling controls metastasis in braf-activated pten-deficient melanomas[J]. Cancer Cell, 2011, 20(6):741-754.doi:10.1016/j.ccr.2011.10.030. pmid: 22172720
[34]	Steinhart Z, Angers S. Wnt signaling in development and tissue homeostasis[J]. Development, 2018, 145(11):dev146589.doi:10.1242/dev.146589.
[35]	Silva-García O, Valdez-Alarcón J J, Baizabal-Aguirre V M. The Wnt/β-catenin signaling pathway controls the inflammatory response in infections caused by pathogenic bacteria[J]. Mediators of Inflammation, 2014, 2014:310183.doi:10.1155/2014/310183.
[36]	Hwang S Y, Deng X M, Byun S, Lee C, Lee S J, Suh H, Zhang J M, Kang Q F, Zhang T, Westover K D, Mandinova A, Lee S W. Direct targeting of β-catenin by a small molecule stimulates proteasomal degradation and suppresses oncogenic Wnt/β-catenin signaling[J]. Cell Reports, 2016, 16(1):28-36.doi:10.1016/j.celrep.2016.05.071. URL
[37]	Zheng X M, Wang S F, Xiao L, Han P P, Xie K K, Ivanovski S, Xiao Y, Zhou Y H. LiCl-induced immunomodulatory periodontal regeneration via the activation of the Wnt/β-catenin signaling pathway[J]. Journal of Periodontal Research, 2022, 57(4):835-848.doi:10.1111/jre.13022. URL

选择文件类型/文献管理软件名称

选择包含的内容

LiCl与MSAB对C₂C₁₂细胞Wnt/β-catenin信号通路及增殖分化的影响

The Effects of LiCl and MSAB on the Wnt/β-Catenin Signaling Pathway, Proliferation,and Differentiation in C₂C₁₂ Cells

RichHTML

PDF

可视化

被引次数

摘要/Abstract

引用本文

使用本文

图/表 6

参考文献 37

相关文章 1

编辑推荐

Metrics

本文评价

模态框（Modal）标题

选择文件类型/文献管理软件名称

选择包含的内容

LiCl与MSAB对C2C12细胞Wnt/β-catenin信号通路及增殖分化的影响

The Effects of LiCl and MSAB on the Wnt/β-Catenin Signaling Pathway, Proliferation,and Differentiation in C2C12 Cells

RichHTML

PDF

可视化

被引次数

摘要/Abstract

引用本文

使用本文

图/表 6

参考文献 37

相关文章 1

编辑推荐

Metrics

本文评价

LiCl与MSAB对C₂C₁₂细胞Wnt/β-catenin信号通路及增殖分化的影响

The Effects of LiCl and MSAB on the Wnt/β-Catenin Signaling Pathway, Proliferation,and Differentiation in C₂C₁₂ Cells