G-Protein-Coupled Receptors Mediate Modulations of Cell Viability and Drug Sensitivity by Aberrantly Expressed Recoverin 3 within A549 Cells
To elucidate the currently unknown molecular mechanisms responsible for the aberrant expression of recoverin (Rec) within cancerous cells, we examined two-dimensional (2D) and three-dimensional (3D) cultures of Rec-negative lung adenocarcinoma A549 cells which had been transfected with a plasmid con...
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MDPI AG
2023-01-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/24/1/771 |
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| author | Hanae Ichioka Yoshihiko Hirohashi Tatsuya Sato Masato Furuhashi Megumi Watanabe Yosuke Ida Fumihito Hikage Toshihiko Torigoe Hiroshi Ohguro |
| author_facet | Hanae Ichioka Yoshihiko Hirohashi Tatsuya Sato Masato Furuhashi Megumi Watanabe Yosuke Ida Fumihito Hikage Toshihiko Torigoe Hiroshi Ohguro |
| author_sort | Hanae Ichioka |
| collection | DOAJ |
| description | To elucidate the currently unknown molecular mechanisms responsible for the aberrant expression of recoverin (Rec) within cancerous cells, we examined two-dimensional (2D) and three-dimensional (3D) cultures of Rec-negative lung adenocarcinoma A549 cells which had been transfected with a plasmid containing human recoverin cDNA (A549 Rec) or an empty plasmid as a mock control (A549 MOCK). Using these cells, we measured cytotoxicity by several anti-tumor agents (2D), cellular metabolism including mitochondrial and glycolytic functions by a Seahorse bio-analyzer (2D), the physical properties, size and stiffness of the 3D spheroids, trypsin sensitivities (2D and 3D), and RNA sequencing analysis (2D). Compared with the A549 MOCK, the A549 Rec cells showed (1) more sensitivity toward anti-tumor agents (2D) and a 0.25% solution of trypsin (3D); (2) a metabolic shift from glycolysis to oxidative phosphorylation; and (3) the formation of larger and stiffer 3D spheroids. RNA sequencing analysis and bioinformatic analyses of the differentially expressed genes (DEGs) using Gene Ontology (GO) enrichment analysis suggested that aberrantly expressed Rec is most likely associated with several canonical pathways including G-protein-coupled receptor (GPCR)-mediated signaling and signaling by the cAMP response element binding protein (CREB). The findings reported here indicate that the aberrantly expressed Rec-induced modulation of the cell viability and drug sensitivity may be GPCR mediated. |
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| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-11T09:58:12Z |
| publishDate | 2023-01-01 |
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| series | International Journal of Molecular Sciences |
| spelling | doaj.art-bf038f45fb6a434487adb4d00858ad4f2023-11-16T15:38:16ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672023-01-0124177110.3390/ijms24010771G-Protein-Coupled Receptors Mediate Modulations of Cell Viability and Drug Sensitivity by Aberrantly Expressed Recoverin 3 within A549 CellsHanae Ichioka0Yoshihiko Hirohashi1Tatsuya Sato2Masato Furuhashi3Megumi Watanabe4Yosuke Ida5Fumihito Hikage6Toshihiko Torigoe7Hiroshi Ohguro8Departments of Ophthalmology, School of Medicine, Sapporo Medical University, Sapporo 060-8556, JapanDepartments of Pathology, School of Medicine, Sapporo Medical University, Sapporo 060-8556, JapanDepartments of Cardiovascular, Renal and Metabolic Medicine, Sapporo Medical University, Sapporo 060-8556, JapanDepartments of Cardiovascular, Renal and Metabolic Medicine, Sapporo Medical University, Sapporo 060-8556, JapanDepartments of Ophthalmology, School of Medicine, Sapporo Medical University, Sapporo 060-8556, JapanDepartments of Ophthalmology, School of Medicine, Sapporo Medical University, Sapporo 060-8556, JapanDepartments of Ophthalmology, School of Medicine, Sapporo Medical University, Sapporo 060-8556, JapanDepartments of Pathology, School of Medicine, Sapporo Medical University, Sapporo 060-8556, JapanDepartments of Ophthalmology, School of Medicine, Sapporo Medical University, Sapporo 060-8556, JapanTo elucidate the currently unknown molecular mechanisms responsible for the aberrant expression of recoverin (Rec) within cancerous cells, we examined two-dimensional (2D) and three-dimensional (3D) cultures of Rec-negative lung adenocarcinoma A549 cells which had been transfected with a plasmid containing human recoverin cDNA (A549 Rec) or an empty plasmid as a mock control (A549 MOCK). Using these cells, we measured cytotoxicity by several anti-tumor agents (2D), cellular metabolism including mitochondrial and glycolytic functions by a Seahorse bio-analyzer (2D), the physical properties, size and stiffness of the 3D spheroids, trypsin sensitivities (2D and 3D), and RNA sequencing analysis (2D). Compared with the A549 MOCK, the A549 Rec cells showed (1) more sensitivity toward anti-tumor agents (2D) and a 0.25% solution of trypsin (3D); (2) a metabolic shift from glycolysis to oxidative phosphorylation; and (3) the formation of larger and stiffer 3D spheroids. RNA sequencing analysis and bioinformatic analyses of the differentially expressed genes (DEGs) using Gene Ontology (GO) enrichment analysis suggested that aberrantly expressed Rec is most likely associated with several canonical pathways including G-protein-coupled receptor (GPCR)-mediated signaling and signaling by the cAMP response element binding protein (CREB). The findings reported here indicate that the aberrantly expressed Rec-induced modulation of the cell viability and drug sensitivity may be GPCR mediated.https://www.mdpi.com/1422-0067/24/1/7713D spheroid culturemelanomaRNA sequencingGene Ontology (GO) enrichment analysisingenuity pathway analysis (IPA)G-protein-coupled receptors |
| spellingShingle | Hanae Ichioka Yoshihiko Hirohashi Tatsuya Sato Masato Furuhashi Megumi Watanabe Yosuke Ida Fumihito Hikage Toshihiko Torigoe Hiroshi Ohguro G-Protein-Coupled Receptors Mediate Modulations of Cell Viability and Drug Sensitivity by Aberrantly Expressed Recoverin 3 within A549 Cells International Journal of Molecular Sciences 3D spheroid culture melanoma RNA sequencing Gene Ontology (GO) enrichment analysis ingenuity pathway analysis (IPA) G-protein-coupled receptors |
| title | G-Protein-Coupled Receptors Mediate Modulations of Cell Viability and Drug Sensitivity by Aberrantly Expressed Recoverin 3 within A549 Cells |
| title_full | G-Protein-Coupled Receptors Mediate Modulations of Cell Viability and Drug Sensitivity by Aberrantly Expressed Recoverin 3 within A549 Cells |
| title_fullStr | G-Protein-Coupled Receptors Mediate Modulations of Cell Viability and Drug Sensitivity by Aberrantly Expressed Recoverin 3 within A549 Cells |
| title_full_unstemmed | G-Protein-Coupled Receptors Mediate Modulations of Cell Viability and Drug Sensitivity by Aberrantly Expressed Recoverin 3 within A549 Cells |
| title_short | G-Protein-Coupled Receptors Mediate Modulations of Cell Viability and Drug Sensitivity by Aberrantly Expressed Recoverin 3 within A549 Cells |
| title_sort | g protein coupled receptors mediate modulations of cell viability and drug sensitivity by aberrantly expressed recoverin 3 within a549 cells |
| topic | 3D spheroid culture melanoma RNA sequencing Gene Ontology (GO) enrichment analysis ingenuity pathway analysis (IPA) G-protein-coupled receptors |
| url | https://www.mdpi.com/1422-0067/24/1/771 |
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