New insights into the roles of the FOXO3 and P27Kip1 genes in signaling pathways
Background: The forkhead box O3 (FOXO3) and p27Kip1 are two important genes in breast cancer progression. In the present study we analyzed the effect of simultaneous FOXO3 silencing and p27Kip1 activation on breast cancer cell survival and the potential targets of these changes in cancer molecular p...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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Upsala Medical Society
2019-07-01
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| Series: | Upsala Journal of Medical Sciences |
| Subjects: | |
| Online Access: | http://dx.doi.org/10.1080/03009734.2019.1623351 |
| _version_ | 1797727075538305024 |
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| author | Sabah Mayahi Masood Golalipour Ahad Yamchi Gagan Deep Jhingan Majid Shahbazi |
| author_facet | Sabah Mayahi Masood Golalipour Ahad Yamchi Gagan Deep Jhingan Majid Shahbazi |
| author_sort | Sabah Mayahi |
| collection | DOAJ |
| description | Background: The forkhead box O3 (FOXO3) and p27Kip1 are two important genes in breast cancer progression. In the present study we analyzed the effect of simultaneous FOXO3 silencing and p27Kip1 activation on breast cancer cell survival and the potential targets of these changes in cancer molecular pathways. Materials and methods: The present study involved the cloning of FOXO3a shRNA and p27Kip1 genes under the control of the bidirectional survivin promoter to down- and up-regulate FOXO3 and p27Kip1 genes, respectively. After transfection of the recombinant expression vector into the breast cancer cell line, the inhibition of cell growth was assessed by MTS and flow cytometry assays. Following the extraction of total mRNA and protein, the expression of target genes was evaluated by qPCR and Western blotting in both treated and untreated cell lines. Then, the downstream protein responses were examined by 2 D electrophoresis. The differentially expressed proteins were also identified by mass spectrometry. Results: Rates of cell proliferation were significantly inhibited in the transfected cell line 72 h post-transfection. Proteomic profiling of the cell line resulted in the identification of seven novel protein markers in breast cancer responsive to these changes in expression of FOXO3 and p27Kip1. The changes in expression of these markers suggested that certain signaling pathways contribute to the development of breast cancer. Conclusion: Simultaneous silencing of FOXO3 and activation of p27Kip1 in MDA-MB-231 cells caused alterations in the expression level of several genes involved in apoptosis, cell proliferation, cell cycle control, tissue invasion, drug resistance, and metastasis. It seems that the identified genes might serve as useful biomarkers for breast cancer. |
| first_indexed | 2024-03-12T10:54:39Z |
| format | Article |
| id | doaj.art-96fc23e81b0b46f8ab7cdcd5ed592d87 |
| institution | Directory Open Access Journal |
| issn | 0300-9734 2000-1967 |
| language | English |
| last_indexed | 2024-03-12T10:54:39Z |
| publishDate | 2019-07-01 |
| publisher | Upsala Medical Society |
| record_format | Article |
| series | Upsala Journal of Medical Sciences |
| spelling | doaj.art-96fc23e81b0b46f8ab7cdcd5ed592d872023-09-02T06:35:05ZengUpsala Medical SocietyUpsala Journal of Medical Sciences0300-97342000-19672019-07-01124314915710.1080/03009734.2019.16233511623351New insights into the roles of the FOXO3 and P27Kip1 genes in signaling pathwaysSabah Mayahi0Masood Golalipour1Ahad Yamchi2Gagan Deep Jhingan3Majid Shahbazi4Golestan University of Medical SciencesGolestan University of Medical SciencesGorgan University of Agricultural Sciences and Natural ResourcesVProteomicsGolestan University of Medical SciencesBackground: The forkhead box O3 (FOXO3) and p27Kip1 are two important genes in breast cancer progression. In the present study we analyzed the effect of simultaneous FOXO3 silencing and p27Kip1 activation on breast cancer cell survival and the potential targets of these changes in cancer molecular pathways. Materials and methods: The present study involved the cloning of FOXO3a shRNA and p27Kip1 genes under the control of the bidirectional survivin promoter to down- and up-regulate FOXO3 and p27Kip1 genes, respectively. After transfection of the recombinant expression vector into the breast cancer cell line, the inhibition of cell growth was assessed by MTS and flow cytometry assays. Following the extraction of total mRNA and protein, the expression of target genes was evaluated by qPCR and Western blotting in both treated and untreated cell lines. Then, the downstream protein responses were examined by 2 D electrophoresis. The differentially expressed proteins were also identified by mass spectrometry. Results: Rates of cell proliferation were significantly inhibited in the transfected cell line 72 h post-transfection. Proteomic profiling of the cell line resulted in the identification of seven novel protein markers in breast cancer responsive to these changes in expression of FOXO3 and p27Kip1. The changes in expression of these markers suggested that certain signaling pathways contribute to the development of breast cancer. Conclusion: Simultaneous silencing of FOXO3 and activation of p27Kip1 in MDA-MB-231 cells caused alterations in the expression level of several genes involved in apoptosis, cell proliferation, cell cycle control, tissue invasion, drug resistance, and metastasis. It seems that the identified genes might serve as useful biomarkers for breast cancer.http://dx.doi.org/10.1080/03009734.2019.1623351breast cancercell signalingfoxo3agene therapyp27kip1proteomics |
| spellingShingle | Sabah Mayahi Masood Golalipour Ahad Yamchi Gagan Deep Jhingan Majid Shahbazi New insights into the roles of the FOXO3 and P27Kip1 genes in signaling pathways Upsala Journal of Medical Sciences breast cancer cell signaling foxo3a gene therapy p27kip1 proteomics |
| title | New insights into the roles of the FOXO3 and P27Kip1 genes in signaling pathways |
| title_full | New insights into the roles of the FOXO3 and P27Kip1 genes in signaling pathways |
| title_fullStr | New insights into the roles of the FOXO3 and P27Kip1 genes in signaling pathways |
| title_full_unstemmed | New insights into the roles of the FOXO3 and P27Kip1 genes in signaling pathways |
| title_short | New insights into the roles of the FOXO3 and P27Kip1 genes in signaling pathways |
| title_sort | new insights into the roles of the foxo3 and p27kip1 genes in signaling pathways |
| topic | breast cancer cell signaling foxo3a gene therapy p27kip1 proteomics |
| url | http://dx.doi.org/10.1080/03009734.2019.1623351 |
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