Abstract P2-06-12: Oncogenic potential of Trefoil factor 3 in initiation of mammary carcinoma through suppression of p53 pathway

Abstract

Abstract Background Oncogenic transformation is a complex multistep process where normal cells acquire the hallmarks of cancer, leading to unrestrained outgrowth of malignant clones. Trefoil Factor 3 (TFF3) is a clinically validated and functionally potent oncogene in mammary carcinoma. Elevated TFF3 expression has been consistently observed in mammary carcinoma, being involved in cancer progression. The present study investigates the potential functional role and the underlying mechanisms of TFF3 in promoting oncogenic transformation early in the onset of mammary carcinoma. Material and method Immortalized human mammary epithelial cells (HMECs): HMEC-hTERT, MCF10A and MCF12A, with forced expression of TFF3, were used as in vitro models and in an orthotopic xenograft model to study the oncogenic roles of TFF3. Furthermore, microarray analysis, immunofluorescence, and ubiquitination and CHX chase assays were used to examine the involvement of p53 pathway in TFF3 mediated-oncogenic transformation. Results Immortalized HMECs with forced expression of TFF3 exhibited the capacity of anchorage independent growth in the soft agar colony formation assay, which is a hallmark of oncogenic transformation. The forced expression of TFF3 also enhanced 3D growth of the immortalized HMECs in matrigel. Furthermore, immortalized HMECs with forced expression of TFF3 gaverise to orthotopic xenograft tumors in nude mice, which are not observed in mice injected with immortalized HMECs. These observations suggest that TFF3 stimulates the oncogenic transformation of non-malignant immortalized HMECs. In addition, the forced expression of TFF3 promoted aberrant cell proliferation, resistance to apoptosis, and increased cell migration and invasion of the HMECs, all these being important hallmarks of cancer. Here, we showed that TFF3-mediated oncogenic transformation of the immortalized HMEC-hTERT cells is dependent on p53 signaling pathway suppression. Mechanistically, TFF3 downregulated NF-κB (p65)-mediated transcription of p53 through decreasing NF-κB (p65) expression and nuclear accumulation. TFF3 also decreased p53 protein levels through post-transcriptional regulation. The forced expression of TFF3 increased MDM2 expression, resulting in an increased ubiquitin-mediated proteasomal degradation of p53. Moreover, forced expression of TFF3 decreased the cleaved form of MDM2, which is responsible for stabilizing p53 protein. Concordantly, HMECs with forced expression of TFF3 exhibited shorter p53 protein half-life as compared to vector control HMECs . Conclusion In summary, our study highlights the oncogenic potential of TFF3 in the initiation of mammary carcinoma through the suppression of the p53 pathway. Citation Format: Chen RM, Pandey V, Chong QY, Poh HM, Zhang MY, Kumar AP, Lobie PE. Oncogenic potential of Trefoil factor 3 in initiation of mammary carcinoma through suppression of p53 pathway [abstract]. In: Proceedings of the 2018 San Antonio Breast Cancer Symposium; 2018 Dec 4-8; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2019;79(4 Suppl):Abstract nr P2-06-12.