A novel protective role for microRNA-3135b in Golgi apparatus fragmentation induced by chemotherapy via GOLPH3/AKT1/mTOR axis in colorectal cancer cells
| Autor: | Rosalío Ramos-Payán, Stephanie I. Nuñez-Olvera, Bibiana Chávez-Munguía, Jonathan Puente-Rivera, Yarely M. Salinas-Vera, Laurence A. Marchat, Alma D. Campos-Parra, María Cruz del Rocío Terrones-Gurrola, Carlos Vázquez-Calzada, Erik Lizárraga-Verdugo, Erika Ruiz-García, César López-Camarillo |
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| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
lcsh:Medicine
Golgi Apparatus Apoptosis Proto-Oncogene Mas Article symbols.namesake Cell Line Tumor Autophagy Humans Fragmentation (cell biology) lcsh:Science 3' Untranslated Regions PI3K/AKT/mTOR pathway Cancer Cell Proliferation Multidisciplinary Chemistry Cell growth TOR Serine-Threonine Kinases lcsh:R Membrane Proteins Transfection Golgi apparatus Cell cycle Cell biology MicroRNAs Oncology Cancer cell symbols lcsh:Q Colorectal Neoplasms Proto-Oncogene Proteins c-akt DNA Damage Signal Transduction |
| Zdroj: | Scientific Reports Scientific Reports, Vol 10, Iss 1, Pp 1-14 (2020) |
| ISSN: | 2045-2322 |
| Popis: | Chemotherapy activates a novel cytoplasmic DNA damage response resulting in Golgi apparatus fragmentation and cancer cell survival. This mechanism is regulated by Golgi phosphoprotein-3 (GOLPH3)/Myo18A/F-actin axis. Analyzing the functions of miR-3135b, a small non-coding RNA with unknown functions, we found that its forced overexpression attenuates the Golgi apparatus fragmentation induced by chemotherapeutic drugs in colorectal cancer (CRC) cells. First, we found that miR-3135b is downregulated in CRC cell lines and clinical tumors. Bioinformatic predictions showed that miR-3135b could be regulating protein-encoding genes involved in cell survival, resistance to chemotherapy, and Golgi dynamics. In agreement, ectopic transfection of miR-3135b in HCT-15 cancer cells significantly inhibited cell proliferation, sensitized cells to 5-fluoruracil (5-FU), and promoted late apoptosis and necrosis. Also, miR-3135b overexpression impaired the cell cycle progression in HCT-15 and SW-480 cancer cells. Because GOLPH3, a gene involved in maintenance of Golgi structure, was predicted as a potential target of miR-3135b, we studied their functional relationships in response to DNA damage induced by chemotherapy. Immunofluorescence and cellular ultrastructure experiments using antibodies against TGN38 protein, a trans-Golgi network marker, showed that 5-FU and doxorubicin treatments result in an apoptosis-independent stacks dispersal of the Golgi ribbon structure in both HCT-15 and SW-480 cells. Remarkably, these cellular effects were dramatically hindered by transfection of miR-3135b mimics. In addition, our functional studies confirmed that miR-3135b binds to the 3′-UTR of GOLPH3 proto-oncogene, and also reduces the levels of p-AKT1 (Ser473) and p-mTOR (Ser2448) signaling transducers, which are key in cell survival and autophagy activation. Moreover, we found that after treatment with 5-FU, TGN38 factor coimmunolocalizes with beclin-1 autophagic protein in discrete structures associated with the fragmented Golgi, suggesting that the activation of pro-survival autophagy is linked to loss of Golgi integrity. These cellular effects in autophagy and Golgi dispersal were reversed by miR-3135b. In summary, we provided experimental evidence suggesting for the first time a novel role for miR-3135b in the protection of chemotherapy-induced Golgi fragmentation via GOLPH3/AKT1/mTOR axis and protective autophagy in colorectal cancer cells. |
| Databáze: | OpenAIRE |
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