Molecular Targeting of HuR Oncoprotein Suppresses MITF and Induces Apoptosis in Melanoma Cells
Treatment of metastatic melanoma possesses challenges due to drug resistance and metastases. Recent advances in targeted therapy and immunotherapy have shown clinical benefits in melanoma patients with increased survival. However, a subset of patients who initially respond to targeted therapy relaps...
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Published in | Cancers Vol. 13; no. 2; p. 166 |
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Main Authors | , , , , , , , |
Format | Journal Article |
Language | English |
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Abstract | Treatment of metastatic melanoma possesses challenges due to drug resistance and metastases. Recent advances in targeted therapy and immunotherapy have shown clinical benefits in melanoma patients with increased survival. However, a subset of patients who initially respond to targeted therapy relapse and succumb to the disease. Therefore, efforts to identify new therapeutic targets are underway. Due to its role in stabilizing several oncoproteins' mRNA, the human antigen R (HuR) has been shown as a promising molecular target for cancer therapy. However, little is known about its potential role in melanoma treatment.
In this study, we tested the impact of siRNA-mediated gene silencing of HuR in human melanoma (MeWo, A375) and normal melanocyte cells in vitro. Cells were treated with HuR siRNA encapsulated in a lipid nanoparticle (NP) either alone or in combination with MEK inhibitor (U0126) and subjected to cell viability, cell-cycle, apoptosis, Western blotting, and cell migration and invasion assays. Cells that were untreated or treated with control siRNA-NP (C-NP) were included as controls.
HuR-NP treatment significantly reduced the expression of HuR and HuR-regulated oncoproteins, induced G1 cell cycle arrest, activated apoptosis signaling cascade, and mitigated melanoma cells' aggressiveness while sparing normal melanocytes. Furthermore, we demonstrated that HuR-NP treatment significantly reduced the expression of the microphthalmia-associated transcription factor (MITF) in both MeWo and MITF-overexpressing MeWo cells (
< 0.05). Finally, combining HuR-NP with U0126 resulted in synergistic antitumor activity against MeWo cells (
< 0.01).
HuR-NP exhibited antitumor activity in melanoma cells independent of their oncogenic B-RAF mutational status. Additionally, combinatorial therapy incorporating MEK inhibitor holds promise in overriding MITF-mediated drug resistance in melanoma. |
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AbstractList | Background: Treatment of metastatic melanoma possesses challenges due to drug resistance and metastases. Recent advances in targeted therapy and immunotherapy have shown clinical benefits in melanoma patients with increased survival. However, a subset of patients who initially respond to targeted therapy relapse and succumb to the disease. Therefore, efforts to identify new therapeutic targets are underway. Due to its role in stabilizing several oncoproteins’ mRNA, the human antigen R (HuR) has been shown as a promising molecular target for cancer therapy. However, little is known about its potential role in melanoma treatment. Methods: In this study, we tested the impact of siRNA-mediated gene silencing of HuR in human melanoma (MeWo, A375) and normal melanocyte cells in vitro. Cells were treated with HuR siRNA encapsulated in a lipid nanoparticle (NP) either alone or in combination with MEK inhibitor (U0126) and subjected to cell viability, cell-cycle, apoptosis, Western blotting, and cell migration and invasion assays. Cells that were untreated or treated with control siRNA-NP (C-NP) were included as controls. Results: HuR-NP treatment significantly reduced the expression of HuR and HuR-regulated oncoproteins, induced G1 cell cycle arrest, activated apoptosis signaling cascade, and mitigated melanoma cells’ aggressiveness while sparing normal melanocytes. Furthermore, we demonstrated that HuR-NP treatment significantly reduced the expression of the microphthalmia-associated transcription factor (MITF) in both MeWo and MITF-overexpressing MeWo cells (p < 0.05). Finally, combining HuR-NP with U0126 resulted in synergistic antitumor activity against MeWo cells (p < 0.01). Conclusion: HuR-NP exhibited antitumor activity in melanoma cells independent of their oncogenic B-RAF mutational status. Additionally, combinatorial therapy incorporating MEK inhibitor holds promise in overriding MITF-mediated drug resistance in melanoma. Treatment of metastatic melanoma possesses challenges due to drug resistance and metastases. Recent advances in targeted therapy and immunotherapy have shown clinical benefits in melanoma patients with increased survival. However, a subset of patients who initially respond to targeted therapy relapse and succumb to the disease. Therefore, efforts to identify new therapeutic targets are underway. Due to its role in stabilizing several oncoproteins' mRNA, the human antigen R (HuR) has been shown as a promising molecular target for cancer therapy. However, little is known about its potential role in melanoma treatment. In this study, we tested the impact of siRNA-mediated gene silencing of HuR in human melanoma (MeWo, A375) and normal melanocyte cells in vitro. Cells were treated with HuR siRNA encapsulated in a lipid nanoparticle (NP) either alone or in combination with MEK inhibitor (U0126) and subjected to cell viability, cell-cycle, apoptosis, Western blotting, and cell migration and invasion assays. Cells that were untreated or treated with control siRNA-NP (C-NP) were included as controls. HuR-NP treatment significantly reduced the expression of HuR and HuR-regulated oncoproteins, induced G1 cell cycle arrest, activated apoptosis signaling cascade, and mitigated melanoma cells' aggressiveness while sparing normal melanocytes. Furthermore, we demonstrated that HuR-NP treatment significantly reduced the expression of the microphthalmia-associated transcription factor (MITF) in both MeWo and MITF-overexpressing MeWo cells ( < 0.05). Finally, combining HuR-NP with U0126 resulted in synergistic antitumor activity against MeWo cells ( < 0.01). HuR-NP exhibited antitumor activity in melanoma cells independent of their oncogenic B-RAF mutational status. Additionally, combinatorial therapy incorporating MEK inhibitor holds promise in overriding MITF-mediated drug resistance in melanoma. |
Author | Ahmed, Rebaz Muralidharan, Ranganayaki Zhao, Yan D Johnston, Sarah E Ekmekcioglu, Suhendan Ramesh, Rajagopal Munshi, Anupama Srivastava, Akhil |
AuthorAffiliation | 3 Stephenson Cancer Center, The University of Oklahoma Health Sciences Center, Oklahoma City, OK 73104, USA; daniel-zhao@ouhsc.edu (Y.D.Z.); anupama-munshi@ouhsc.edu (A.M.) 6 Department of Radiation Oncology, The University of Oklahoma Health Sciences Center, Oklahoma City, OK 73104, USA 1 Department of Pathology, The University of Oklahoma Health Sciences Center, Oklahoma City, OK 73104, USA; rebaz-ahmed@ouhsc.edu (R.A.); mranga550@gmail.com (R.M.); akhil-srivastava@ouhsc.edu (A.S.) 2 Graduate Program in Biomedical Sciences, The University of Oklahoma Health Sciences Center, Oklahoma City, OK 73104, USA 5 Department of Melanoma Medical Oncology, The University of Texas MD Anderson Cancer Center, Houston, TX 77030, USA; sekmekcioglu@mdanderson.org 4 Department of Biostatistics and Epidemiology, The University of Oklahoma Health Sciences Center, Oklahoma City, OK 73104, USA; sarah-johnston@ouhsc.edu |
AuthorAffiliation_xml | – name: 4 Department of Biostatistics and Epidemiology, The University of Oklahoma Health Sciences Center, Oklahoma City, OK 73104, USA; sarah-johnston@ouhsc.edu – name: 3 Stephenson Cancer Center, The University of Oklahoma Health Sciences Center, Oklahoma City, OK 73104, USA; daniel-zhao@ouhsc.edu (Y.D.Z.); anupama-munshi@ouhsc.edu (A.M.) – name: 1 Department of Pathology, The University of Oklahoma Health Sciences Center, Oklahoma City, OK 73104, USA; rebaz-ahmed@ouhsc.edu (R.A.); mranga550@gmail.com (R.M.); akhil-srivastava@ouhsc.edu (A.S.) – name: 2 Graduate Program in Biomedical Sciences, The University of Oklahoma Health Sciences Center, Oklahoma City, OK 73104, USA – name: 5 Department of Melanoma Medical Oncology, The University of Texas MD Anderson Cancer Center, Houston, TX 77030, USA; sekmekcioglu@mdanderson.org – name: 6 Department of Radiation Oncology, The University of Oklahoma Health Sciences Center, Oklahoma City, OK 73104, USA |
Author_xml | – sequence: 1 givenname: Rebaz surname: Ahmed fullname: Ahmed, Rebaz organization: Graduate Program in Biomedical Sciences, The University of Oklahoma Health Sciences Center, Oklahoma City, OK 73104, USA – sequence: 2 givenname: Ranganayaki surname: Muralidharan fullname: Muralidharan, Ranganayaki organization: Stephenson Cancer Center, The University of Oklahoma Health Sciences Center, Oklahoma City, OK 73104, USA – sequence: 3 givenname: Akhil orcidid: 0000-0003-2015-259X surname: Srivastava fullname: Srivastava, Akhil organization: Stephenson Cancer Center, The University of Oklahoma Health Sciences Center, Oklahoma City, OK 73104, USA – sequence: 4 givenname: Sarah E surname: Johnston fullname: Johnston, Sarah E organization: Department of Biostatistics and Epidemiology, The University of Oklahoma Health Sciences Center, Oklahoma City, OK 73104, USA – sequence: 5 givenname: Yan D surname: Zhao fullname: Zhao, Yan D organization: Department of Biostatistics and Epidemiology, The University of Oklahoma Health Sciences Center, Oklahoma City, OK 73104, USA – sequence: 6 givenname: Suhendan orcidid: 0000-0003-4079-6632 surname: Ekmekcioglu fullname: Ekmekcioglu, Suhendan organization: Department of Melanoma Medical Oncology, The University of Texas MD Anderson Cancer Center, Houston, TX 77030, USA – sequence: 7 givenname: Anupama surname: Munshi fullname: Munshi, Anupama organization: Department of Radiation Oncology, The University of Oklahoma Health Sciences Center, Oklahoma City, OK 73104, USA – sequence: 8 givenname: Rajagopal orcidid: 0000-0002-7658-5338 surname: Ramesh fullname: Ramesh, Rajagopal organization: Stephenson Cancer Center, The University of Oklahoma Health Sciences Center, Oklahoma City, OK 73104, USA |
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CitedBy_id | crossref_primary_10_1182_bloodadvances_2023011132 crossref_primary_10_3390_cancers14030707 crossref_primary_10_1016_j_addr_2021_114068 crossref_primary_10_1016_j_addr_2021_113918 crossref_primary_10_1016_j_addr_2021_114088 crossref_primary_10_1155_2023_6882851 crossref_primary_10_1002_jcp_30311 crossref_primary_10_1007_s12094_023_03109_5 crossref_primary_10_1016_j_semcancer_2022_06_008 crossref_primary_10_3390_microorganisms12020314 crossref_primary_10_1002_jcp_31229 crossref_primary_10_1158_0008_5472_CAN_23_0972 |
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Keywords | metastases targeted therapy nanoparticles HuR melanoma MITF siRNA |
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Snippet | Treatment of metastatic melanoma possesses challenges due to drug resistance and metastases. Recent advances in targeted therapy and immunotherapy have shown... Background: Treatment of metastatic melanoma possesses challenges due to drug resistance and metastases. Recent advances in targeted therapy and immunotherapy... |
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SubjectTerms | Antitumor activity Apoptosis Cancer therapies Cell cycle Cell growth Cell migration Cell viability Cytotoxicity Drug resistance Experiments Gene silencing HuR protein Immunotherapy Kinases Lipids MEK inhibitors Melanocytes Melanoma Metastases Metastasis Microphthalmia-associated transcription factor Nanoparticles Oncoproteins Patients Proteins Raf protein siRNA Skin cancer Western blotting |
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Title | Molecular Targeting of HuR Oncoprotein Suppresses MITF and Induces Apoptosis in Melanoma Cells |
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