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Cited by 8Glioma Diagnosis and TreatmentCancer Cells and MetastasisMicrotubule and mitosis dynamics

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TUBB2B regulates epithelial-mesenchymal transition via interaction with Vimentin to promote glioma migration and invasion

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Li, Junxi, Zhou, Zhengjun, Zhang, Junrong, Wang, Ming, Luan, Xingzhao, Zhao, Mingkuan, Jiang, Geng, Wang, Guiyuan, Li, Shenjie, Xiang, Wei, Chen, Ligang, Zhou, Jie·2024·Cancer Cell International
DOI 10.1186/s12935-024-03618-5PMC PMC11662515

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Abstract

BACKGROUND: Epithelial-mesenchymal transition (EMT) plays a crucial role in the migration and invasion capabilities of glioblastoma (GBM) cells. Several studies have established tubulin as a significant regulator of the EMT process. Tubulin beta 2B class IIb (TUBB2B), a critical component of microtubules, has been linked to the prognosis of various tumors. However, the specific biological function and mechanism of TUBB2B in GBM remain unclear. METHODS: In vitro experiments demonstrated that TUBB2B knockdown inhibited the migration and invasion of GBM cells, while its overexpression enhanced these capabilities. Western blot, immunofluorescence (IF) and co-immunoprecipitation (Co-IP) assays revealed that TUBB2B interacts with Vimentin. Molecular docking and residue mutation scanning indicated that TUBB2B interacts with Vimentin at the R391/K392/A393/F394 sites. In vivo experiments using nude mice confirmed that TUBB2B knockdown inhibited GBM cell invasion and migration. RESULTS: TUBB2B was upregulated in GBM tissue samples compared with normal tissues. The sites of TUBB2B(R391/K392/A393/F394) physically interacts with Vimentin to induce EMT, which promotes migration and invasion. CONCLUSION: TUBB2B may regulate EMT and promote the migration and invasion of GBM cells through its interaction with Vimentin, highlighting TUBB2B as a potential therapeutic target for GBM.

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Fig. 1 TUBB2B is upregulated in GBM tissues. (A-B) Based on GEPIA database analysis, TUBB2B was significantly upregulated in LGG and GBM; P < 0.05. (C-E) Prognostic value of TUBB2B in TGGA, CGGA, Rembrandt datasets. (F) Immunohistochemical detection of TUBB2B protein levels in glioma tissues and normal brain tissues (Scale bar, 200 μm). (G) Expression of TUBB2B in 8 normal tissues and 80 GBM tissues. (H) Kaplan Meier analysis was used to evaluate the overall survival of GBM patients based on TUBB2B expression (n = 64, **P = 0.0073). All results are displayed as the mean ± SD. p < 0.05, *; p < 0.01, **; p < 0.001, ***
Fig. 1 TUBB2B is upregulated in GBM tissues. (A-B) Based on GEPIA database analysis, TUBB2B was significantly upregulated in LGG and GBM; P < 0.05. (C-E) Prognostic value of TUBB2B in TGGA, CGGA, Rembrandt datasets. (F) Immunohistochemical detection of TUBB2B protein levels in glioma tissues and normal brain tissues (Scale bar, 200 μm). (G) Expression of TUBB2B in 8 normal tissues and 80 GBM tissues. (H) Kaplan Meier analysis was used to evaluate the overall survival of GBM patients based on TUBB2B expression (n = 64, **P = 0.0073). All results are displayed as the mean ± SD. p < 0.05, *; p < 0.01, **; p < 0.001, ***CC BY-NC-ND · PubMed Central
Fig. 2 Effect of TUBB2B knockdown and overexpression on the proliferation and migration of glioma cells. (A) Expression levels of TUBB2B in glioma cell lines and astrocyte cell lines. (B) Western blot analysis of TUBB2B expression. (C) Cell migration and invasion were assessed in GBM cells expressing TUBB2B shRNA using Transwell assay (Scale bar, 100 μm). (D) The cell migration ability of GBM cells expressing TUBB2B shRNA was detected by wound healing assay (Scale bar, 200 μm). (E) The efficiency of TUBB2B overexpression after transfection with TUBB2B OERNA were evaluated by qRT-PCR. (F) Western blot analysis of TUBB2B expression. (G) Cell migration and invasion were assessed in GBM cells expressing TUBB2B OERNA using Transwell assay (Scale bar, 100 μm). (H) The cell migration ability of GBM cells expressing TUBB2B OERNA was detected by wound healing assay (Scale bar, 200 μm). All results are displayed as the mean ± SD. p < 0.05, *; p < 0.01, **; p < 0.001, ***; p < 0.0001, ****
Fig. 2 Effect of TUBB2B knockdown and overexpression on the proliferation and migration of glioma cells. (A) Expression levels of TUBB2B in glioma cell lines and astrocyte cell lines. (B) Western blot analysis of TUBB2B expression. (C) Cell migration and invasion were assessed in GBM cells expressing TUBB2B shRNA using Transwell assay (Scale bar, 100 μm). (D) The cell migration ability of GBM cells expressing TUBB2B shRNA was detected by wound healing assay (Scale bar, 200 μm). (E) The efficiency of TUBB2B overexpression after transfection with TUBB2B OERNA were evaluated by qRT-PCR. (F) Western blot analysis of TUBB2B expression. (G) Cell migration and invasion were assessed in GBM cells expressing TUBB2B OERNA using Transwell assay (Scale bar, 100 μm). (H) The cell migration ability of GBM cells expressing TUBB2B OERNA was detected by wound healing assay (Scale bar, 200 μm). All results are displayed as the mean ± SD. p < 0.05, *; p < 0.01, **; p < 0.001, ***; p < 0.0001, ****CC BY-NC-ND · PubMed Central
Fig. 3 Transcriptome analysis of LN229 cell line. (A) KEGG enrichment analysis of the RNA-seq results. (B) GO enrichment analysis of the RNA-seq results. (C) The PPI network of TUBB2B and Vimentin (scorevalue ≥ 0.3). (D) RNA-seq expression of heatmaps of genes involved in extracellular matrix, focal adhesion, and cell migration
Fig. 3 Transcriptome analysis of LN229 cell line. (A) KEGG enrichment analysis of the RNA-seq results. (B) GO enrichment analysis of the RNA-seq results. (C) The PPI network of TUBB2B and Vimentin (scorevalue ≥ 0.3). (D) RNA-seq expression of heatmaps of genes involved in extracellular matrix, focal adhesion, and cell migrationCC BY-NC-ND · PubMed Central
Fig. 4 TUBB2B promotes EMT in GBM cells (A) The levels of EMT markers and TUBB2B in the LN229 and T98 cells. (B) Representative morphology of LN229 and T98 cells silenced for TUBB2B expression (Scale bar, 50 μm). (C) Representative images (20 × magnification) of immunofluorescence staining patterns for nucleus (DAPI, blue), Actin(green), and Tubulin (red) evaluated in control (NC) and shRNA cells (Scale bar, 20 μm)
Fig. 4 TUBB2B promotes EMT in GBM cells (A) The levels of EMT markers and TUBB2B in the LN229 and T98 cells. (B) Representative morphology of LN229 and T98 cells silenced for TUBB2B expression (Scale bar, 50 μm). (C) Representative images (20 × magnification) of immunofluorescence staining patterns for nucleus (DAPI, blue), Actin(green), and Tubulin (red) evaluated in control (NC) and shRNA cells (Scale bar, 20 μm)CC BY-NC-ND · PubMed Central
Fig. 5 TUBB2B interacts with Vimentin in GBM cells (A) STRING database protein interaction network of TUBB2B. (B) The interaction between TUBB2B and Vimentin inLN229 and T98 cells was determined using Co-IP. (C) Double staining of GBM cells with TUBB2B and Vimentin was assessed using confocal microscopy (Scale bar, 10 μm. Pearson’s R value = 0.60). (D) The predicted binding mode of TUBB2B and Vimentin, as well as the detail interactions between two proteins. (E) Schematic representation of Flag-TUBB2B-NC and the Flag-TUBB2B mutants. All results are displayed as the mean ± SD. p < 0.05, *; p < 0.01, **; p < 0.001, ***; p < 0.0001, ****
Fig. 5 TUBB2B interacts with Vimentin in GBM cells (A) STRING database protein interaction network of TUBB2B. (B) The interaction between TUBB2B and Vimentin inLN229 and T98 cells was determined using Co-IP. (C) Double staining of GBM cells with TUBB2B and Vimentin was assessed using confocal microscopy (Scale bar, 10 μm. Pearson’s R value = 0.60). (D) The predicted binding mode of TUBB2B and Vimentin, as well as the detail interactions between two proteins. (E) Schematic representation of Flag-TUBB2B-NC and the Flag-TUBB2B mutants. All results are displayed as the mean ± SD. p < 0.05, *; p < 0.01, **; p < 0.001, ***; p < 0.0001, ****CC BY-NC-ND · PubMed Central
Fig. 6 TUBB2B-Mut4 inhibits GBM cell migration and invasion in vivo. (A) Migration and invasion ability of TUBB2B-Mut4 in GBM cells (Scale bar, 10 μm). (B) The cell migration ability of GBM cells expressing TUBB2B-Mut4 was detected by wound healing assay (Scale bar, 10 μm). All results are displayed as the mean ± SD. p < 0.05, *; p < 0.01, **; p < 0.001, ***
Fig. 6 TUBB2B-Mut4 inhibits GBM cell migration and invasion in vivo. (A) Migration and invasion ability of TUBB2B-Mut4 in GBM cells (Scale bar, 10 μm). (B) The cell migration ability of GBM cells expressing TUBB2B-Mut4 was detected by wound healing assay (Scale bar, 10 μm). All results are displayed as the mean ± SD. p < 0.05, *; p < 0.01, **; p < 0.001, ***CC BY-NC-ND · PubMed Central
Fig. 7 Effect of tubb2b knockdown on GBM cell migration and invasion in vivo. (A) In vivo invasion assay for NC and ShTUBB2B LN229 cells (Scale bar = 200 μm). (B) Overall survival of mice. (C) IHC analysis of TUBB2B, Vimentin and N-cadherin levels in mice brain tumors (Scale bar, 50 μm). (D)Relative expression levels of TUBB2B, Vimentin and N-cadherin in IHC. All results are displayed as the mean ± SD. p < 0.05, *; p < 0.01, **
Fig. 7 Effect of tubb2b knockdown on GBM cell migration and invasion in vivo. (A) In vivo invasion assay for NC and ShTUBB2B LN229 cells (Scale bar = 200 μm). (B) Overall survival of mice. (C) IHC analysis of TUBB2B, Vimentin and N-cadherin levels in mice brain tumors (Scale bar, 50 μm). (D)Relative expression levels of TUBB2B, Vimentin and N-cadherin in IHC. All results are displayed as the mean ± SD. p < 0.05, *; p < 0.01, **CC BY-NC-ND · PubMed Central

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Research works printed but not linked: 38

  1. Rivera M, Bander ED, Cisse B. Perspectives on Microglia-based Immune therapies against Glioblastoma. World Neurosurg. 2021;154:228–31. · DOI
  2. Weller M, van den Bent M, Preusser M, Le Rhun E, Tonn JC, Minniti G, et al. EANO guidelines on the diagnosis and treatment of diffuse gliomas of adulthood. Nat Reviews Clin Oncol. 2021;18(3):170–86. · DOI · Paper page
  3. Nørøxe DS, Poulsen HS, Lassen U. Hallmarks of glioblastoma: a systematic review. ESMO Open 2016;1(6). · DOI
  4. Yang J, Antin P, Berx G, Blanpain C, Brabletz T, Bronner M, et al. Guidelines and definitions for research on epithelial–mesenchymal transition. Nat Rev Mol Cell Biol. 2020;21(6):341–52. · DOI · Paper page
  5. Chi M, Liu J, Mei C, Shi Y, Liu N, Jiang X et al. TEAD4 functions as a prognostic biomarker and triggers EMT via PI3K/AKT pathway in bladder cancer. J Experimental Clin Cancer Res. 2022;41(1). · DOI
  6. Aiello NM, Kang Y. Context-dependent EMT programs in cancer metastasis. J Exp Med. 2019;216(5):1016–26. · DOI
  7. Zeisberg M, Neilson EG. Biomarkers for epithelial-mesenchymal transitions. J Clin Investig. 2009;119(6):1429–37. · DOI · Paper page
  8. Shibue T, Brooks MW, Weinberg RA. An integrin-linked machinery of cytoskeletal regulation that enables experimental tumor initiation and metastatic colonization. Cancer Cell. 2013;24(4):481–98. · DOI
  9. McConkey DJ, Choi W, Marquis L, Martin F, Williams MB, Shah J, et al. Role of epithelial-to-mesenchymal transition (EMT) in drug sensitivity and metastasis in bladder cancer. Cancer Metastasis Rev. 2009;28(3):335–44. · DOI
  10. Admoni-Elisha L, Elbaz T, Chopra A, Shapira G, Bedford Mark T, Fry Christopher J, et al. TWIST1 methylation by SETD6 selectively antagonizes LINC-PINT expression in glioma. Nucleic Acids Res. 2022;50(12):6903–18. · DOI
  11. Colas E, Pedrola N, Devis L, Ertekin T, Campoy I, Martínez E, et al. The EMT signaling pathways in endometrial carcinoma. Clin Transl Oncol. 2012;14(10):715–20. · DOI
  12. van Zijl F, Zulehner G, Petz M, Schneller D, Kornauth C, Hau M, et al. Epithelial–mesenchymal transition in hepatocellular carcinoma. Future Oncol. 2009;5(8):1169–79. · DOI
  13. Bates RC, Mercurio A. The epithelial-mesenchymal tansition (EMT) and colorectal cancer progression. Cancer Biol Ther. 2005;4(4):371–6. · DOI
  14. Babu D, Mudiraj A, Yadav N, Y.B.V.K C, Panigrahi M, Prakash Babu P. Rabeprazole has efficacy per se and reduces resistance to temozolomide in glioma via EMT inhibition. Cell Oncol. 2021;44(4):889–905. · DOI
  15. Ruksha K, Mezheyeuski A, Nerovnya A, Bich T, Tur G, Gorgun J et al. Over-expression of βII-Tubulin and especially its localization in cell nuclei correlates with poorer outcomes in Colorectal Cancer. Cells. 2019;8(1). · DOI
  16. Wang X, Shi J, Huang M, Chen J, Dan J, Tang Y et al. TUBB2B facilitates progression of hepatocellular carcinoma by regulating cholesterol metabolism through targeting HNF4A/CYP27A1. Cell Death Dis. 2023;14(3). · DOI
  17. Liotti A, La Civita E, Cennamo M, Crocetto F, Ferro M, Guadagno E, et al. Periprostatic adipose tissue promotes prostate cancer resistance to docetaxel by paracrine IGF-1 upregulation of TUBB2B beta‐tubulin isoform. Prostate. 2021;81(7):407–17. · DOI
  18. Wang Z, Wang H, Wang Z, He S, Jiang Z, Yan C, et al. Associated analysis of PER1/TUBB2B with endometrial cancer development caused by circadian rhythm disorders. Med Oncol. 2020;37(10):90. · DOI
  19. Maillard C, Roux CJ, Charbit-Henrion F, Steffann J, Laquerriere A, Quazza F, et al. Tubulin mutations in human neurodevelopmental disorders. Semin Cell Dev Biol. 2023;137:87–95. · DOI
  20. Kozakov D, Hall DR, Xia B, Porter KA, Padhorny D, Yueh C, et al. The ClusPro web server for protein–protein docking. Nat Protoc. 2017;12(2):255–78. · DOI
  21. Schymkowitz J, Borg J, Stricher F, Nys R, Rousseau F, Serrano L. The FoldX web server: an online force field. Nucleic Acids Res. 2005;33(suppl2):W382–8. · DOI · Paper page
  22. Sadrkhanloo M, Entezari M, Orouei S, Ghollasi M, fathi N, Rezaei S, et al. STAT3-EMT axis in tumors: modulation of cancer metastasis, stemness and therapy response. Pharmacol Res. 2022;182:106311. · DOI
  23. Kar R, Jha NK, Jha SK, Sharma A, Dholpuria S, Asthana N et al. A NOTCH deeper into the Epithelial-To-Mesenchymal transition (EMT) program in breast Cancer. Genes. 2019;10(12). · DOI
  24. Xu J, Lamouille S, Derynck R. TGF-β-induced epithelial to mesenchymal transition. Cell Res. 2009;19(2):156–72. · DOI · Paper page
  25. Atiya HI, Gorecki G, Garcia GL, Frisbie LG, Baruwal R, Coffman L. Stromal-modulated epithelial-to-mesenchymal transition in Cancer cells. Biomolecules [Internet]. 2023; 13(11). · DOI
  26. Lin Y-T, Wu K-J. Epigenetic regulation of epithelial-mesenchymal transition: focusing on hypoxia and TGF-β signaling. J Biomed Sci. 2020;27(1):39. · DOI
  27. Stanton RA, Gernert KM, Nettles JH, Aneja R. Drugs that target dynamic microtubules: a new molecular perspective. Med Res Rev. 2011;31(3):443–81. · DOI
  28. Wen J, Wang Q, Zhang W, Wang W. TUBA1A licenses APC/C-mediated mitotic progression to drive glioblastoma growth by inhibiting PLK3. FEBS Lett. 2023;597(24):3072–86. · DOI
  29. Sobierajska K, Wieczorek K, Ciszewski WM, Sacewicz-Hofman I, Wawro ME, Wiktorska M, et al. β-III tubulin modulates the behavior of snail overexpressed during the epithelial-to-mesenchymal transition in colon cancer cells. Biochimica et Biophysica Acta (BBA) -. Mol Cell Res. 2016;1863(9):2221–33.
  30. Gui S, Chen P, Liu Y, Chen Q, Cheng T, Lv S, et al. TUBA1C expression promotes proliferation by regulating the cell cycle and indicates poor prognosis in glioma. Biochem Biophys Res Commun. 2021;577:130–8. · DOI
  31. Liu J, Li Y. Upregulation of MAPK10, TUBB2B and RASL11B may contribute to the development of neuroblastoma. Mol Med Rep. 2019. · DOI
  32. Strouhalova K, Přechová M, Gandalovičová A, Brábek J, Gregor M, Rosel D. Vimentin intermediate filaments as potential target for Cancer Treatment. Cancers [Internet]. 2020; 12(1). · DOI
  33. Sannino G, Marchetto A, Kirchner T, Grünewald TGP. Epithelial-to-mesenchymal and mesenchymal-to-epithelial transition in mesenchymal tumors: a Paradox in Sarcomas? Cancer Res. 2017;77(17):4556–61. · DOI
  34. Huang X, Xiang L, Wang B, Hu J, Liu C, Ren A et al. CMTM6 promotes migration, invasion, and EMT by interacting with and stabilizing vimentin in hepatocellular carcinoma cells. J Translational Med. 2021;19(1). · DOI
  35. Preitner N, Quan J, Nowakowski Dan W, Hancock Melissa L, Shi J, Tcherkezian J, et al. APC is an RNA-Binding protein, and its Interactome provides a link to neural development and Microtubule Assembly. Cell. 2014;158(2):368–82. · DOI
  36. Lu Q, Luduena RF. Removal of β III isotype enhances Taxol Induced Microtubule Assembly. Cell Struct Funct. 1993;18(3):173–82. · DOI
  37. Hendrix MJ, Seftor EA, Seftor RE, Trevor KT. Experimental co-expression of vimentin and keratin intermediate filaments in human breast cancer cells results in phenotypic interconversion and increased invasive behavior. Am J Pathol. 1997;150(2):483–95.
  38. Liu C-Y, Lin H-H, Tang M-J, Wang Y-K. Vimentin contributes to epithelial-mesenchymal transition cancer cell mechanics by mediating cytoskeletal organization and focal adhesion maturation. Oncotarget; Vol 6, No 18. 2015. · DOI

Metadata

Authors
Li, J., Zhou, Z., Zhang, J., Wang, M., Luan, X., Zhao, M., Jiang, G., Wang, G., Li, S., Xiang, W., Chen, L., & Zhou, J.
Affiliations
Southwest Medical University (CN)
Corresponding author
  • Xiang, Wei
  • Chen, Ligang
  • Zhou, Jie
ORCID
Venue
Cancer Cell International
Publisher
Springer Science and Business Media LLC
Published
2024-12-20
Volume
24
Issue
1
Pages
423-423
ISSN
1475-2867 (ISSN-L)
DOI
10.1186/s12935-024-03618-5
PMC
PMC11662515
OpenAlex
W4405636637
Cited by
8
References
1
Citations by year
2026: 6 · 2025: 2
FWCI
2.25
Type
journal article
Language
en
Open access
Licence
Locations
  • Cancer Cell International (publishedVersion, cc-by-nc-nd)
  • Research Square (acceptedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by-nc-nd)
  • PubMed (publishedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion)
Funders
  • The research was supported by National Natural Science Foundation project (82072780 and 82372825)
  • the Science and Technology Department of Sichuan Province (2022YFS0630)
  • the Sichuan Medical Youth Innovation Research Project (Q22044)
  • the Luzhou Science and Technology Program (2023JYJ028)
  • the University Science and Technology Strategic Cooperation Project (2024SNXNYD03)
Topics
  • Health Sciences › Medicine › Oncology › Glioma Diagnosis and Treatment
  • Health Sciences › Medicine › Oncology › Cancer Cells and Metastasis
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Microtubule and mitosis dynamics
Keywords
TUBB2B, epithelial-mesenchymal transition, vimentin, glioblastoma, cell migration, tumor invasion, protein-protein interaction, microtubules, therapeutic target

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Li, J., Zhou, Z., Zhang, J., Wang, M., Luan, X., Zhao, M., Jiang, G., Wang, G., Li, S., Xiang, W., Chen, L., & Zhou, J. (2024). TUBB2B regulates epithelial-mesenchymal transition via interaction with Vimentin to promote glioma migration and invasion. Cancer Cell International. https://doi.org/10.1186/s12935-024-03618-5