Abstract 952: Attenuation of β-Catenin signaling by ormeloxifene in hepatocellular carcinoma

Abstract

Abstract Background: Hepatocellular carcinoma (HCC) is the most frequent type of primary liver cancer with approximately 41,000 new diagnosis and 29,000 fatalities every year only in the U.S. Severe toxicity and poor response to available chemotherapeutic agents make HCC clinical management extremely difficult. Aberrant β-catenin signaling is found to play a critical role in differentiation grades of HCC patients. Thus, attempts to find more potent and non-toxic β-Catenin inhibitors is urgently needed for cancer therapy. Notably, ormeloxifene (ORM), clinically approved selective estrogen receptor modulator with therapeutic index, has shown excellent anticancer activity, rendering it as an ideal candidate for repurposing. In this study, we investigated the anti-cancer potential of ORM against hepatocellular carcinoma. Methodology: Cell proliferation and colony formation assays were performed to assess the therapeutic activity of ORM in human hepatocellular carcinoma (HepG2, SK-HEP-1, Hep3B, and C3A) cells. Boyden chamber and Matrigel assays were carried out for investigating the effect of ORM on migration and invasion abilities of HCC cells, respectively. The effects of ORM on β-catenin and EMT associated proteins were analyzed through Western blotting and qPCR. Confocal microscopy was used to determine the β-catenin nuclear localization following ORM treatment in HCC cells. Results: As compared to vehicle-treated group, ORM treatment (2.5-20 µM) suppressed proliferation and colony formation in human hepatocellular carcinoma cells in a dose and time-dependent manner. Moreover, ORM treatment suppresses the migration and invasion of human hepatocellular carcinoma cells as shown by wound healing and Matrigel invasion assay, respectively. ORM effectively inhibited the protein levels and mRNA expression of total β-catenin. Additionally, our confocal microscopy results further showed reduced nuclear translocation of β-catenin following the ORM treatment. ORM treatment inhibited epithelial-to-mesenchymal transition (EMT) process as evident by repression of N-cadherin, Slug, Snail, vimentin, MMPs (MMP2 and MMP3), and induced the expression of pGSK3β. Experiments are also being conducted to see how ORM affects epigenetic markers linked to EMT and β-catenin signaling. Conclusion: Taken together, ORM inhibited β-catenin signaling and exhibited potent anticancer effects against HCC and could be investigated further as a novel therapeutic modality for HCC treatment. Citation Format: Mohammed Sikander, Shabnam Malik, Anyssa Rodriguez, Molly Vela, Daniel Zubieta, Vivek K. Kashyap, Anupam Dhasmana, Bilal B. Hafeez, Sheema Khan, Fathi T. Halaweish, Subhash C. Chauhan, Meena Jaggi. Attenuation of β-Catenin signaling by ormeloxifene in hepatocellular carcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 952.