Posters
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Post-doc
Discipline/Specialty (Author 1)
Cancer and Immunology
Academic Level (Author 2)
Faculty
Discipline/Specialty (Author 2)
Cancer and Immunology
Academic Level (Author 5)
Faculty
Discipline/Specialty (Author 5)
Cancer and Immunology
Discipline/Specialty (Author 6)
Cancer and Immunology
Academic Level (Author 7)
Faculty
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Cancer and Immunology
Discipline Track
Biomedical Science
Abstract
Background: Hepatocellular carcinoma (HCC) is one of the deadliest cancers, with a five-year survival rate. While Sorafenib (SRB) is the standard therapy, it is only effective in prolonging survival for some patients, emphasizing the urgent need for alternative treatment options. Resistance to apoptosis is a defining characteristic of cancer cells and a crucial factor in cancer recurrence and relapse. Although, anticancer potential of the FDA-approved compound Ormeloxifene (ORM), which has well-defined pharmacokinetic and pharmacodynamic properties, has been explored in a variety of cancers, its efficacy in HCC has remained unknown. Therefore, the purpose of this study was to investigate the role and molecular mechanisms underlying ORM in SRB-induced apoptosis in HCC.
Methods: In this study, we investigated the efficacy of ORM in hepatocellular carcinoma cell (HepG2, Hep3B, C3a, and SKHep-1) lines alone, and in combination with SRB. Cell proliferation was assessed using MTT and xCELLigence assays. The impact of ORM on the clonogenic potential of HCC cells was evaluated through colony formation assays. The effect of ORM on inducing apoptosis was determined using Annexin V assays. Cell migration was assessed using Boyden chambers, and cell invasion was evaluated with Matrigel invasion chambers. Subsequent to treatment in hepatocellular carcinoma cells, Western blotting, quantitative polymerase chain reaction (qPCR), and immunofluorescence studies were conducted to investigate the effects of ORM, SRB alone, and/or combination on pro-apoptotic, anti-apoptotic, EMT, and associated signaling effector proteins.
Results: By employing MTT and xCELLigence assays, our study showed that ORM induces dose- and time-dependent inhibition of cell proliferation in hepatocellular cancer (HepG2, Hep3B, C3a, and SKHep-1) cells at various concentrations (0, 5, 10, and 15 µM). The combination of ORM and SRB together had a significant synergistic inhibitory effect on HCC cells. Additionally, compared to ORM and SRB treatment alone, Annexin V staining showed that combined treatment of ORM and SRB significantly enhanced the apoptosis induction in hepatocellular carcinoma cells. Mechanistically, the expression of anti-apoptotic markers like Mcl-1, Bcl-2, and Bcl-xl is significantly reduced when ORM and SRB are combined as evidenced by Western blotting. Similarly, the inhibition of colony formation, invasion, and migration of hepatocellular carcinoma cells following co-treatment was significantly higher as compared to either treatment alone. In addition, the expression of EMT markers like N-cadherin, snail, vimentin, and MMPs in HCC cells is down-regulated following combined treatment compared to ORM and SRB alone.
Conclusion: ORM demonstrates promising chemo-therapeutic and chemo-sensitizing effects in the treatment of hepatocellular carcinoma, offering potential as a novel therapeutic strategy for this aggressive cancer.
Presentation Type
Poster
Recommended Citation
Malik, Shabnam; Sikander, Mohammed; Zubieta, Daniel; Rodriguez, Anyssa; Yallapu, Murali; Jaggi, Meena; and Chauhan, Subhash C., "Ormeloxifene Suppresses EMT in Hepatocellular Carcinoma by Targeting β-catenin Signaling Pathway" (2024). Research Colloquium. 64.
https://scholarworks.utrgv.edu/colloquium/2024/posters/64
Ormeloxifene Suppresses EMT in Hepatocellular Carcinoma by Targeting β-catenin Signaling Pathway
Background: Hepatocellular carcinoma (HCC) is one of the deadliest cancers, with a five-year survival rate. While Sorafenib (SRB) is the standard therapy, it is only effective in prolonging survival for some patients, emphasizing the urgent need for alternative treatment options. Resistance to apoptosis is a defining characteristic of cancer cells and a crucial factor in cancer recurrence and relapse. Although, anticancer potential of the FDA-approved compound Ormeloxifene (ORM), which has well-defined pharmacokinetic and pharmacodynamic properties, has been explored in a variety of cancers, its efficacy in HCC has remained unknown. Therefore, the purpose of this study was to investigate the role and molecular mechanisms underlying ORM in SRB-induced apoptosis in HCC.
Methods: In this study, we investigated the efficacy of ORM in hepatocellular carcinoma cell (HepG2, Hep3B, C3a, and SKHep-1) lines alone, and in combination with SRB. Cell proliferation was assessed using MTT and xCELLigence assays. The impact of ORM on the clonogenic potential of HCC cells was evaluated through colony formation assays. The effect of ORM on inducing apoptosis was determined using Annexin V assays. Cell migration was assessed using Boyden chambers, and cell invasion was evaluated with Matrigel invasion chambers. Subsequent to treatment in hepatocellular carcinoma cells, Western blotting, quantitative polymerase chain reaction (qPCR), and immunofluorescence studies were conducted to investigate the effects of ORM, SRB alone, and/or combination on pro-apoptotic, anti-apoptotic, EMT, and associated signaling effector proteins.
Results: By employing MTT and xCELLigence assays, our study showed that ORM induces dose- and time-dependent inhibition of cell proliferation in hepatocellular cancer (HepG2, Hep3B, C3a, and SKHep-1) cells at various concentrations (0, 5, 10, and 15 µM). The combination of ORM and SRB together had a significant synergistic inhibitory effect on HCC cells. Additionally, compared to ORM and SRB treatment alone, Annexin V staining showed that combined treatment of ORM and SRB significantly enhanced the apoptosis induction in hepatocellular carcinoma cells. Mechanistically, the expression of anti-apoptotic markers like Mcl-1, Bcl-2, and Bcl-xl is significantly reduced when ORM and SRB are combined as evidenced by Western blotting. Similarly, the inhibition of colony formation, invasion, and migration of hepatocellular carcinoma cells following co-treatment was significantly higher as compared to either treatment alone. In addition, the expression of EMT markers like N-cadherin, snail, vimentin, and MMPs in HCC cells is down-regulated following combined treatment compared to ORM and SRB alone.
Conclusion: ORM demonstrates promising chemo-therapeutic and chemo-sensitizing effects in the treatment of hepatocellular carcinoma, offering potential as a novel therapeutic strategy for this aggressive cancer.