Mir-675-5p supports hypoxia-induced drug resistance in colorectal cancer cells
Abstract Background The uncontrolled proliferation of cancer cells determines hypoxic conditions within the neoplastic mass with consequent activation of specific molecular pathways that allow cells to survive despite oxygen deprivation. The same molecular pathways are often the cause of chemoresist...
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| Format: | Article |
| Language: | English |
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BMC
2022-05-01
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| Series: | BMC Cancer |
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| Online Access: | https://doi.org/10.1186/s12885-022-09666-2 |
| _version_ | 1811341081850150912 |
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| author | Chiara Zichittella Maria Magdalena Barreca Aurora Cordaro Chiara Corrado Riccardo Alessandro Alice Conigliaro |
| author_facet | Chiara Zichittella Maria Magdalena Barreca Aurora Cordaro Chiara Corrado Riccardo Alessandro Alice Conigliaro |
| author_sort | Chiara Zichittella |
| collection | DOAJ |
| description | Abstract Background The uncontrolled proliferation of cancer cells determines hypoxic conditions within the neoplastic mass with consequent activation of specific molecular pathways that allow cells to survive despite oxygen deprivation. The same molecular pathways are often the cause of chemoresistance. This study aims to investigate the role of the hypoxia-induced miR-675-5p in 5-Fluorouracil (5-FU) resistance on colorectal cancer (CRC) cells. Methods CRC cell lines were treated with 5-Fu and incubated in normoxic or hypoxic conditions; cell viability has been evaluated by MTT assay. MiR-675-5p levels were analysed by RT-PCR and loss and gain expression of the miRNA has been obtained by the transfection of miRNA antagomir or miRNA mimic. Total protein expression of different apoptotic markers was analysed through western blot assay. MirWalk 2.0 database search engine was used to investigate the putative targets of the miR-675-5p involved in the apoptotic process. Finally, the luciferase assay was done to confirm Caspase-3 as a direct target of the miR-675-5p. Results Our data demonstrated that hypoxia-induced miR-675-5p counteracts the apoptotic signal induced by 5-FU, thus taking part in the drug resistance response. We showed that the apoptotic markers, cleaved PARP and cleaved caspase-3, increased combining miR-675-5p inhibition with 5-FU treatment. Moreover, we identified pro-caspase-3 among the targets of the miR-675-5p. Conclusion Our data demonstrate that the inhibition of hypoxia-induced miR-675-5p combined with 5-FU treatment can enhances drug efficacy in both prolonged hypoxia and normoxia, indicating a possible strategy to partially overcome chemoresistance. |
| first_indexed | 2024-04-13T18:52:22Z |
| format | Article |
| id | doaj.art-0c1f7c181f654042b7abbd69dc97e970 |
| institution | Directory Open Access Journal |
| issn | 1471-2407 |
| language | English |
| last_indexed | 2024-04-13T18:52:22Z |
| publishDate | 2022-05-01 |
| publisher | BMC |
| record_format | Article |
| series | BMC Cancer |
| spelling | doaj.art-0c1f7c181f654042b7abbd69dc97e9702022-12-22T02:34:23ZengBMCBMC Cancer1471-24072022-05-0122111210.1186/s12885-022-09666-2Mir-675-5p supports hypoxia-induced drug resistance in colorectal cancer cellsChiara Zichittella0Maria Magdalena Barreca1Aurora Cordaro2Chiara Corrado3Riccardo Alessandro4Alice Conigliaro5Department of Biomedicine, Neuroscience and Advanced Diagnostics (Bi.N.D.), Section of Biology and Genetics, University of PalermoDepartment of Biomedicine, Neuroscience and Advanced Diagnostics (Bi.N.D.), Section of Biology and Genetics, University of PalermoDepartment of Biomedicine, Neuroscience and Advanced Diagnostics (Bi.N.D.), Section of Biology and Genetics, University of PalermoDepartment of Biomedicine, Neuroscience and Advanced Diagnostics (Bi.N.D.), Section of Biology and Genetics, University of PalermoDepartment of Biomedicine, Neuroscience and Advanced Diagnostics (Bi.N.D.), Section of Biology and Genetics, University of PalermoDepartment of Biomedicine, Neuroscience and Advanced Diagnostics (Bi.N.D.), Section of Biology and Genetics, University of PalermoAbstract Background The uncontrolled proliferation of cancer cells determines hypoxic conditions within the neoplastic mass with consequent activation of specific molecular pathways that allow cells to survive despite oxygen deprivation. The same molecular pathways are often the cause of chemoresistance. This study aims to investigate the role of the hypoxia-induced miR-675-5p in 5-Fluorouracil (5-FU) resistance on colorectal cancer (CRC) cells. Methods CRC cell lines were treated with 5-Fu and incubated in normoxic or hypoxic conditions; cell viability has been evaluated by MTT assay. MiR-675-5p levels were analysed by RT-PCR and loss and gain expression of the miRNA has been obtained by the transfection of miRNA antagomir or miRNA mimic. Total protein expression of different apoptotic markers was analysed through western blot assay. MirWalk 2.0 database search engine was used to investigate the putative targets of the miR-675-5p involved in the apoptotic process. Finally, the luciferase assay was done to confirm Caspase-3 as a direct target of the miR-675-5p. Results Our data demonstrated that hypoxia-induced miR-675-5p counteracts the apoptotic signal induced by 5-FU, thus taking part in the drug resistance response. We showed that the apoptotic markers, cleaved PARP and cleaved caspase-3, increased combining miR-675-5p inhibition with 5-FU treatment. Moreover, we identified pro-caspase-3 among the targets of the miR-675-5p. Conclusion Our data demonstrate that the inhibition of hypoxia-induced miR-675-5p combined with 5-FU treatment can enhances drug efficacy in both prolonged hypoxia and normoxia, indicating a possible strategy to partially overcome chemoresistance.https://doi.org/10.1186/s12885-022-09666-2Colorectal cancer (CRC)HypoxiaMicroRNADrug resistanceApoptosis5-fluorouracil (5-FU) |
| spellingShingle | Chiara Zichittella Maria Magdalena Barreca Aurora Cordaro Chiara Corrado Riccardo Alessandro Alice Conigliaro Mir-675-5p supports hypoxia-induced drug resistance in colorectal cancer cells BMC Cancer Colorectal cancer (CRC) Hypoxia MicroRNA Drug resistance Apoptosis 5-fluorouracil (5-FU) |
| title | Mir-675-5p supports hypoxia-induced drug resistance in colorectal cancer cells |
| title_full | Mir-675-5p supports hypoxia-induced drug resistance in colorectal cancer cells |
| title_fullStr | Mir-675-5p supports hypoxia-induced drug resistance in colorectal cancer cells |
| title_full_unstemmed | Mir-675-5p supports hypoxia-induced drug resistance in colorectal cancer cells |
| title_short | Mir-675-5p supports hypoxia-induced drug resistance in colorectal cancer cells |
| title_sort | mir 675 5p supports hypoxia induced drug resistance in colorectal cancer cells |
| topic | Colorectal cancer (CRC) Hypoxia MicroRNA Drug resistance Apoptosis 5-fluorouracil (5-FU) |
| url | https://doi.org/10.1186/s12885-022-09666-2 |
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