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. 2022 Jul;11(7):2205-2216.
doi: 10.21037/tcr-21-2413.

GLIPR1 regulates the TIMP1-CD63-ITGB1-AKT signaling pathway in glioma cells and induces malignant transformation of astroglioma

Feng Li  1 , Weifeng Zhang  1 , Ming Wang  1 , Pifeng Jia  1
Affiliations

GLIPR1 regulates the TIMP1-CD63-ITGB1-AKT signaling pathway in glioma cells and induces malignant transformation of astroglioma

Feng Li et al. Transl Cancer Res. 2022 Jul.

Abstract

Background: Astrocytoma (ACM) is characterized by high recurrence rate, high mortality, and extremely poor clinical prognosis. The new diagnostic and prognostic tumor markers related to ACM was found to improve the diagnosis rate and reduce the poor prognosis.

Methods: The activity of SHC-44 and SW1783 cells under the regulation of glioma pathogenesis-related protein 1 (GLIPR1) was investigated by CCK8 analysis. The effect of GLIPR1 on the proliferation of SHC-44 and SW1783 cells was analyzed by cell colony-forming experiment. The migration of SHC-44 and SW1783 cells under the regulation of GLIPR1 was analyzed by transwell assay. The effects of GLIPR1 on the invasion and migration of SHC-44 and SW1783 cells were analyzed by cell scratch test and transwell assay. Immunofluorescence and Co-IP assays were employed to analyze the expression characteristics of GLIPR1 and CD63 proteins. The effect of GLIPR1 on the protein expression of GLIPR1, TIMP1, CD63, ITGB1, and AKT in SHC-44 and SW1783 cells was analyzed by western blot. The effect of anti-AKT on the protein expression of GLIPR1, TIMP1, CD63, ITGB1, and AKT in SHC-44 and SW1783 cells was performed by western blot.

Results: The outcomes revealed that GLIPR1 could enhance the activity, proliferation, migration, and invasion of ACM cells, which might be associated with the activation of the TIMP1-CD63-ITGB1-AKT signaling pathway.

Conclusions: Taken together, GLIPR1 might be a potential target for the prevention or management of ACM in the clinic.

Keywords: Glioma pathogenesis-related protein 1 (GLIPR1); TIMP1-CD63-ITGB1-AKT; astroglioma; invasion; migration.

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Conflict of interest statement

Conflicts of Interest: All authors have completed the ICMJE uniform disclosure form (available at https://tcr.amegroups.com/article/view/10.21037/tcr-21-2413/coif). The authors have no conflicts of interest to declare.

Figures

Figure 1
Figure 1
GLIPR1 affected the proliferation of ACM cells. (A) The effect of GLIPR1 on the proliferation of SHC-44 cells at different culture times (0, 24, 36, and 48 h). (B) The effect of GLIPR1 on the proliferation of SHC-44 cells at different culture times (0, 24, 36, and 48 h). (*) and (#) represent a significance compared with the Model and NC group, respectively. *, P<0.05; **, P<0.01; ##, P<0.01. NC, normal group (SVG p12 cells); Model, model group. ACM, astrocytoma.
Figure 2
Figure 2
GLIPR1 induced the proliferation of SHC-44 and SW1783 cells by cell colony-forming assay. (A) The effect of overexpression or interference with GLIPR1 on the proliferation of SHC-44 cells. (B) The effect of overexpression or interference with GLIPR1 on the proliferation of SW1783 cells. (*) represents a significance compared with the Model group; **, P<0.01. NC, normal group; Model, model group.
Figure 3
Figure 3
GLIPR1 enhanced the invasion of SHC-44 and SW1783 cells by transwell assay, and cells were stained with crystal violet. (A) Effect of overexpression or interference with GLIPR1 on the invasion of SHC-44 cells. (B) Effect of overexpression or interference with GLIPR1 on the invasion of SW1783 cells. Magnification: ×200, scale bar: 100 µm. (*) represents a significance compared with the Model group. **, P<0.01. NC, normal group; Model, model group.
Figure 4
Figure 4
GLIPR1 improved the migration of SHC-44 and SW1783 cells. (A) Phase-contrast images of scratch assay of SHC-44 cells at 0 and 24 h after wound and quantification of the area covered. (B) Phase-contrast images of scratch assay of SW1783 cells at 0 and 24 h after wound and quantification of the area covered. (*) represents a significance compared with the Model group. ****, P<0.0001. NC, normal group; Model, model group.
Figure 5
Figure 5
GLIPR1 increased cell migration of SHC-44 and SW1783 cells by verified transwell assay, and cells were stained with crystal violet. (A) Overexpression or interference with GLIPR1 further increased the invasion of SHC-44 cells. (B) Overexpression or interference with GLIPR1 further increased the invasion of SW1783 cells. Magnification: ×200, scale bar: 100 µm. (*) represents a significance compared with the Model group. ****, P<0.0001. NC, normal group; Model, model group.
Figure 6
Figure 6
GLIPR1 regulated the CD63 expression by the interaction between them. (A) The protein expressions of GLIPR1 and CD63 in SHC-44 cells were observed by immunofluorescence assay. (B) The protein expressions of GLIPR1 and CD63 in SW1783 cells were observed by immunofluorescence assay. Red dots referred to GLIPR1 proteins, green dots referred to CD63 proteins, and blue dots referred to living cells. Scale bar: 100 µm. NC, normal group; Model, model group.
Figure 7
Figure 7
Co-IP assay was performed to detect the binding of GLIPR1 to CD63 in SHC-44 (A) and SW1783 (B) cells. NC, normal group; Model, model group.
Figure 8
Figure 8
GLIPR1 activated the TIMP1-CD63-ITGB1-AKT signaling pathway. Protein levels of GLIPR1, TIMP1, CD63, ITGB1, and AKT in SHC-44 (A) and SW1783 (B) cells were detected by western blot. The relative expression of GLIPR1, TIMP1, CD63, ITGB1, and AKT was calculated via normalization to β-actin expression. (*) represents a significance compared with the Model group. *, P<0.05; **, P<0.01; ***, P<0.001; ****, P<0.0001. NC, normal group; Model, model group.
Figure 9
Figure 9
TIMP1-CD63-ITGB1-AKT signaling pathway played a key role in GLIPR1 activation. Protein levels of GLIPR1, TIMP1, CD63, ITGB1, and AKT in SHC-44 (A) and SW1783 (B) cells were detected by western blot after treatment by anti-AKT. The relative expression of GLIPR1, TIMP1, CD63, ITGB1, and AKT was calculated via normalization to β-actin expression. (*) represents a significance compared with the Model group. ***, P<0.001; ****, P<0.0001. NC, normal group; Model, model group.
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References

    1. Jiang T, Mao Y, Ma W, et al. CGCG clinical practice guidelines for the management of adult diffuse gliomas. Cancer Lett 2016;375:263-73. 10.1016/j.canlet.2016.01.024 - DOI - PubMed
    1. Bush NA, Chang SM, Berger MS. Current and future strategies for treatment of glioma. Neurosurg Rev 2017;40:1-14. 10.1007/s10143-016-0709-8 - DOI - PubMed
    1. Walker DG, Kaye AH. Diagnosis and management of astrocytomas, oligodendrogliomas and mixed gliomas: a review. Australas Radiol 2001;45:472-82. 10.1046/j.1440-1673.2001.00959.x - DOI - PubMed
    1. Mathieu D, Fortin D. The role of chemotherapy in the treatment of malignant astrocytomas. Can J Neurol Sci 2006;33:127-40. 10.1017/S0317167100004881 - DOI - PubMed
    1. Okamoto Y, Di Patre PL, Burkhard C, et al. Population-based study on incidence, survival rates, and genetic alterations of low-grade diffuse astrocytomas and oligodendrogliomas. Acta Neuropathol 2004;108:49-56. 10.1007/s00401-004-0861-z - DOI - PubMed