CLEC19A overexpression inhibits tumor cell proliferation/migration and promotes apoptosis concomitant suppression of PI3K/AKT/NF-κB signaling pathway in glioblastoma multiforme
Abstract Background GBM is the most frequent malignant primary brain tumor in humans. The CLEC19A is a member of the C-type lectin family, which has a high expression in brain tissue. Herein, we sought to carry out an in-depth analysis to pinpoint the role of CLEC19A expression in GBM. Methods To de...
| Main Authors: | , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
BMC
2024-01-01
|
| Series: | BMC Cancer |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s12885-023-11755-9 |
| _version_ | 1797363192742019072 |
|---|---|
| author | Fatemeh Mohajerani Zahra Moazezi Tehrankhah Saeid Rahmani Nastaran Afsordeh Sajad Shafiee Mohammad Hossein Pourgholami Bahram M. Soltani Majid Sadeghizadeh |
| author_facet | Fatemeh Mohajerani Zahra Moazezi Tehrankhah Saeid Rahmani Nastaran Afsordeh Sajad Shafiee Mohammad Hossein Pourgholami Bahram M. Soltani Majid Sadeghizadeh |
| author_sort | Fatemeh Mohajerani |
| collection | DOAJ |
| description | Abstract Background GBM is the most frequent malignant primary brain tumor in humans. The CLEC19A is a member of the C-type lectin family, which has a high expression in brain tissue. Herein, we sought to carry out an in-depth analysis to pinpoint the role of CLEC19A expression in GBM. Methods To determine the localization of CLEC19A, this protein was detected using Western blot, Immunocytochemistry/Immunofluorescence, and confocal microscopy imaging. CLEC19A expression in glioma cells and tissues was evaluated by qRT-PCR. Cell viability, proliferation, migration, and apoptosis were examined through MTT assay, CFSE assay, colony formation, wound healing assay, transwell test, and flow cytometry respectively after CLEC19A overexpression. The effect of CLEC19A overexpression on the PI3K/AKT/NF-κB signaling pathway was investigated using Western blot. An in vivo experiment substantiated the in vitro results using the glioblastoma rat models. Results Our in-silico analysis using TCGA data and measuring CLEC19A expression level by qRT-PCR determined significantly lower expression of CLEC19A in human glioma tissues compared to healthy brain tissues. By employment of ICC/IF, confocal microscopy imaging, and Western blot we could show that CLEC19A is plausibly a secreted protein. Results obtained from several in vitro readouts showed that CLEC19A overexpression in U87 and C6 glioma cell lines is associated with the inhibition of cell proliferation, viability, and migration. Further, qRT-PCR and Western blot analysis showed CLEC19A overexpression could reduce the expression levels of PI3K, VEGFα, MMP2, and NF-κB and increase PTEN, TIMP3, RECK, and PDCD4 expression levels in glioma cell lines. Furthermore, flow cytometry results revealed that CLEC19A overexpression was associated with significant cell cycle arrest and promotion of apoptosis in glioma cell lines. Interestingly, using a glioma rat model we could substantiate that CLEC19A overexpression suppresses glioma tumor growth. Conclusions To our knowledge, this is the first report providing in-silico, molecular, cellular, and in vivo evidences on the role of CLEC19A as a putative tumor suppressor gene in GBM. These results enhance our understanding of the role of CLEC19A in glioma and warrant further exploration of CLEC19A as a potential therapeutic target for GBM. |
| first_indexed | 2024-03-08T16:17:40Z |
| format | Article |
| id | doaj.art-c1b896286efb4851b6f069c0d49980c8 |
| institution | Directory Open Access Journal |
| issn | 1471-2407 |
| language | English |
| last_indexed | 2024-03-08T16:17:40Z |
| publishDate | 2024-01-01 |
| publisher | BMC |
| record_format | Article |
| series | BMC Cancer |
| spelling | doaj.art-c1b896286efb4851b6f069c0d49980c82024-01-07T12:30:14ZengBMCBMC Cancer1471-24072024-01-0124112010.1186/s12885-023-11755-9CLEC19A overexpression inhibits tumor cell proliferation/migration and promotes apoptosis concomitant suppression of PI3K/AKT/NF-κB signaling pathway in glioblastoma multiformeFatemeh Mohajerani0Zahra Moazezi Tehrankhah1Saeid Rahmani2Nastaran Afsordeh3Sajad Shafiee4Mohammad Hossein Pourgholami5Bahram M. Soltani6Majid Sadeghizadeh7Department of Genetics, Faculty of Biological Sciences, Tarbiat Modares UniversityDepartment of Genetics, Faculty of Biological Sciences, Tarbiat Modares UniversitySchool of Computer Science, Institute for Research in Fundamental Sciences (IPM)Department of Physiology, Faculty of Medical Sciences, Tarbiat Modares UniversityDepartment of Neurosurgery, Mazandaran University of Medical SciencesDepartment of Physiology, Faculty of Medical Sciences, Tarbiat Modares UniversityDepartment of Genetics, Faculty of Biological Sciences, Tarbiat Modares UniversityDepartment of Genetics, Faculty of Biological Sciences, Tarbiat Modares UniversityAbstract Background GBM is the most frequent malignant primary brain tumor in humans. The CLEC19A is a member of the C-type lectin family, which has a high expression in brain tissue. Herein, we sought to carry out an in-depth analysis to pinpoint the role of CLEC19A expression in GBM. Methods To determine the localization of CLEC19A, this protein was detected using Western blot, Immunocytochemistry/Immunofluorescence, and confocal microscopy imaging. CLEC19A expression in glioma cells and tissues was evaluated by qRT-PCR. Cell viability, proliferation, migration, and apoptosis were examined through MTT assay, CFSE assay, colony formation, wound healing assay, transwell test, and flow cytometry respectively after CLEC19A overexpression. The effect of CLEC19A overexpression on the PI3K/AKT/NF-κB signaling pathway was investigated using Western blot. An in vivo experiment substantiated the in vitro results using the glioblastoma rat models. Results Our in-silico analysis using TCGA data and measuring CLEC19A expression level by qRT-PCR determined significantly lower expression of CLEC19A in human glioma tissues compared to healthy brain tissues. By employment of ICC/IF, confocal microscopy imaging, and Western blot we could show that CLEC19A is plausibly a secreted protein. Results obtained from several in vitro readouts showed that CLEC19A overexpression in U87 and C6 glioma cell lines is associated with the inhibition of cell proliferation, viability, and migration. Further, qRT-PCR and Western blot analysis showed CLEC19A overexpression could reduce the expression levels of PI3K, VEGFα, MMP2, and NF-κB and increase PTEN, TIMP3, RECK, and PDCD4 expression levels in glioma cell lines. Furthermore, flow cytometry results revealed that CLEC19A overexpression was associated with significant cell cycle arrest and promotion of apoptosis in glioma cell lines. Interestingly, using a glioma rat model we could substantiate that CLEC19A overexpression suppresses glioma tumor growth. Conclusions To our knowledge, this is the first report providing in-silico, molecular, cellular, and in vivo evidences on the role of CLEC19A as a putative tumor suppressor gene in GBM. These results enhance our understanding of the role of CLEC19A in glioma and warrant further exploration of CLEC19A as a potential therapeutic target for GBM.https://doi.org/10.1186/s12885-023-11755-9CLEC19AC-type lectinGlioblastomaPI3K/AKT/NF-κB pathwayTumor suppressor gene |
| spellingShingle | Fatemeh Mohajerani Zahra Moazezi Tehrankhah Saeid Rahmani Nastaran Afsordeh Sajad Shafiee Mohammad Hossein Pourgholami Bahram M. Soltani Majid Sadeghizadeh CLEC19A overexpression inhibits tumor cell proliferation/migration and promotes apoptosis concomitant suppression of PI3K/AKT/NF-κB signaling pathway in glioblastoma multiforme BMC Cancer CLEC19A C-type lectin Glioblastoma PI3K/AKT/NF-κB pathway Tumor suppressor gene |
| title | CLEC19A overexpression inhibits tumor cell proliferation/migration and promotes apoptosis concomitant suppression of PI3K/AKT/NF-κB signaling pathway in glioblastoma multiforme |
| title_full | CLEC19A overexpression inhibits tumor cell proliferation/migration and promotes apoptosis concomitant suppression of PI3K/AKT/NF-κB signaling pathway in glioblastoma multiforme |
| title_fullStr | CLEC19A overexpression inhibits tumor cell proliferation/migration and promotes apoptosis concomitant suppression of PI3K/AKT/NF-κB signaling pathway in glioblastoma multiforme |
| title_full_unstemmed | CLEC19A overexpression inhibits tumor cell proliferation/migration and promotes apoptosis concomitant suppression of PI3K/AKT/NF-κB signaling pathway in glioblastoma multiforme |
| title_short | CLEC19A overexpression inhibits tumor cell proliferation/migration and promotes apoptosis concomitant suppression of PI3K/AKT/NF-κB signaling pathway in glioblastoma multiforme |
| title_sort | clec19a overexpression inhibits tumor cell proliferation migration and promotes apoptosis concomitant suppression of pi3k akt nf κb signaling pathway in glioblastoma multiforme |
| topic | CLEC19A C-type lectin Glioblastoma PI3K/AKT/NF-κB pathway Tumor suppressor gene |
| url | https://doi.org/10.1186/s12885-023-11755-9 |
| work_keys_str_mv | AT fatemehmohajerani clec19aoverexpressioninhibitstumorcellproliferationmigrationandpromotesapoptosisconcomitantsuppressionofpi3kaktnfkbsignalingpathwayinglioblastomamultiforme AT zahramoazezitehrankhah clec19aoverexpressioninhibitstumorcellproliferationmigrationandpromotesapoptosisconcomitantsuppressionofpi3kaktnfkbsignalingpathwayinglioblastomamultiforme AT saeidrahmani clec19aoverexpressioninhibitstumorcellproliferationmigrationandpromotesapoptosisconcomitantsuppressionofpi3kaktnfkbsignalingpathwayinglioblastomamultiforme AT nastaranafsordeh clec19aoverexpressioninhibitstumorcellproliferationmigrationandpromotesapoptosisconcomitantsuppressionofpi3kaktnfkbsignalingpathwayinglioblastomamultiforme AT sajadshafiee clec19aoverexpressioninhibitstumorcellproliferationmigrationandpromotesapoptosisconcomitantsuppressionofpi3kaktnfkbsignalingpathwayinglioblastomamultiforme AT mohammadhosseinpourgholami clec19aoverexpressioninhibitstumorcellproliferationmigrationandpromotesapoptosisconcomitantsuppressionofpi3kaktnfkbsignalingpathwayinglioblastomamultiforme AT bahrammsoltani clec19aoverexpressioninhibitstumorcellproliferationmigrationandpromotesapoptosisconcomitantsuppressionofpi3kaktnfkbsignalingpathwayinglioblastomamultiforme AT majidsadeghizadeh clec19aoverexpressioninhibitstumorcellproliferationmigrationandpromotesapoptosisconcomitantsuppressionofpi3kaktnfkbsignalingpathwayinglioblastomamultiforme |