We are analyzing https://link.springer.com/article/10.1186/s13046-018-0880-6.

Title:
HSP90AA1-mediated autophagy promotes drug resistance in osteosarcoma | Journal of Experimental & Clinical Cancer Research
Description:
Background Osteosarcoma is the most common primary bone tumor in children and adolescents. Unfortunately, osteosarcoma treatments often fail due to the development of chemoresistance, of which the underlying molecular mechanisms still remain unclear. In this study, we demonstrated that HSP90AA1 gene is responsible for drug resistance in osteosarcoma through an autophagy-related mechanism. Methods shRNAs were transfected into osteosarcoma cells for knockdown of HSP90AA1 gene. Stable HSP90AA1 overexpressing osteosarcoma cell lines were obtained by lentivirus infection. mRNA and protein expressions of HSP90AA1 in osteosarcoma cells were tested by quantitative real-time PCR and western blot, respectively. Autophagy of osteosarcoma cells was detected by western blot of LC3, transmission electron microscopy and fluorescence microscope. mRFP-GFP-LC3 lentiviral transfection was also performed to detect autophagic flux. NOD/SCID mices were inoculated with MG-63 tumor cells transfected with HSP90AA1 specific shRNA. TUNEL and LC3 staining were performed to detect apoptosis and autophagy of resected tumor tissues. Results Doxorubicin, cisplatin, and methotrexate, which are commonly used in chemotherapy, each induced HSP90AA1 upregulation in human osteosarcoma cells. Suppression of HSP90AA1 restored the sensitivity of osteosarcoma cells to chemotherapy both in vivo and in vitro. Mechanism study indicated that autophagy is responsible for the chemoresistance in osteosarcoma cells. HSP90AA1 increased drug resistance by inducing autophagy and inhibiting apoptosis. Suppression of HSP90AA1 diminished autophagic protection in response to chemotherapy in osteosarcoma cells. Moreover, HSP90AA1 promotes autophagy through PI3K/Akt/mTOR pathway and inhibits apoptosis through JNK/P38 pathway. Conclusion We showed that chemotherapy agents can induce HSP90AA1 expression in osteosarcoma cells. And HSP90AA1, acting as an important regulator of autophagy, is a critical factor in the development of osteosarcoma chemoresistance both in vitro and in vivo. HSP90AA1 provides a novel therapeutic target for improving osteosarcoma treatment.
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cells, hspaa, autophagy, osteosarcoma, cell, pubmed, article, apoptosis, chemotherapy, fig, cancer, google, scholar, shrna, cas, control, resistance, treatment, pathway, doxorubicin, treated, drug, protein, transfected, expression, tumor, additional, western, blot, group, knockdown, role, dox, μgml, analyzed, results, stress, pikaktmtor, file, figure, analysis, study, autophagic, increased, survival, central, promotes, cisplatin, china, versus,

Topics {✒️}

annexin v-pe/propidium iodide annexin v-pe/pi staining lc3-ii-positive punctate pattern pi3k/akt/mtor signaling pathway mrfp-gfp-lc3 lentiviral transfection full size image quantitative real-time pcr control shrna/hsp90aa1 shrna pi3k/akt/mtor signaling article download pdf transfected control/hsp90aa1 shrna targeting hmgb1-mediated autophagy ceramide/akt/mtor pathway phblv control/hsp90a1 lentivirus pi3k/akt/mtor pathway mrfp-gfp-lc3 construct hsp90aa1-overexpressing mg-63 cells cell stress-induced alterations hsp90aa1-mediated antiapoptotic effect mrfp-gfp-lc3 lentivirus multi-factor involved process autophagy-mediated cell survival human liver cancer mitogen-activated protein kinases p62/sqstm1-mediated degradation pi3k/akt signaling full access breast cancer res control shrna transfection human osteosarcoma cells state key laboratory military medical university subcutaneous tumor size hsp90aa1 inhibited chemotherapy-induced related subjects promote chemotherapy-induced autophagy control/hsp90aa1 shrna mammalian autophagy research drug resistance mediated o'sullivan gc zheng guo medical ethics committee hsp90aa1 shrna transfection privacy choices/manage cookies p38 kinase pathways hsp90aa1-mediated resistance prosthetic bone replacement cell signaling technology osteosarcoma cell growth hsp90aa1 shrna group

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Implication of heat shock protein 90 (HSP90) in tumor angiogenesis: a molecular target for anti-angiogenic therapy?

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         description:Osteosarcoma is the most common primary bone tumor in children and adolescents. Unfortunately, osteosarcoma treatments often fail due to the development of chemoresistance, of which the underlying molecular mechanisms still remain unclear. In this study, we demonstrated that HSP90AA1 gene is responsible for drug resistance in osteosarcoma through an autophagy-related mechanism. shRNAs were transfected into osteosarcoma cells for knockdown of HSP90AA1 gene. Stable HSP90AA1 overexpressing osteosarcoma cell lines were obtained by lentivirus infection. mRNA and protein expressions of HSP90AA1 in osteosarcoma cells were tested by quantitative real-time PCR and western blot, respectively. Autophagy of osteosarcoma cells was detected by western blot of LC3, transmission electron microscopy and fluorescence microscope. mRFP-GFP-LC3 lentiviral transfection was also performed to detect autophagic flux. NOD/SCID mices were inoculated with MG-63 tumor cells transfected with HSP90AA1 specific shRNA. TUNEL and LC3 staining were performed to detect apoptosis and autophagy of resected tumor tissues. Doxorubicin, cisplatin, and methotrexate, which are commonly used in chemotherapy, each induced HSP90AA1 upregulation in human osteosarcoma cells. Suppression of HSP90AA1 restored the sensitivity of osteosarcoma cells to chemotherapy both in vivo and in vitro. Mechanism study indicated that autophagy is responsible for the chemoresistance in osteosarcoma cells. HSP90AA1 increased drug resistance by inducing autophagy and inhibiting apoptosis. Suppression of HSP90AA1 diminished autophagic protection in response to chemotherapy in osteosarcoma cells. Moreover, HSP90AA1 promotes autophagy through PI3K/Akt/mTOR pathway and inhibits apoptosis through JNK/P38 pathway. We showed that chemotherapy agents can induce HSP90AA1 expression in osteosarcoma cells. And HSP90AA1, acting as an important regulator of autophagy, is a critical factor in the development of osteosarcoma chemoresistance both in vitro and in vivo. HSP90AA1 provides a novel therapeutic target for improving osteosarcoma treatment.
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