Abstract PO5-24-08: Differential exploitation of Cation Transport Regulator homolog 1 (CHAC1) by Wild-type and mutant p53

Abstract

Abstract Background: Cancer cells are constantly exposed to harsh conditions like nutrient deprivation, hypoxia, and calcium imbalance leading to the induction of UPR. In its early phase, UPR aims to restore homeostasis while prolonged stress leads to induction of apoptosis. Thus, it is not surprising to observe that expression of the UPR components has been reported to both promote as well as suppress tumorigenesis. Studies have shown that Cation transport regulator homolog 1 (CHAC1) plays a dual role in cancer progression. The objective of the present study is to investigate the p53 mediated regulation of CHAC1 in breast cancer. Methods: We initially subjected a breast cancer tissue microarray (TMA) to immunohistochemistry (IHC) with p53 and CHAC1 specific antibody. We further did in vitro experiments like western blotting, qRT-PCR to see the expression of CHAC1 when wild type (WT) p53 was either induced using camptothecin (CPT) and Nutlin-3 in WT p53 expressing breast cancer cells or by the ectopic WT and mutant p53 transfection in null p53 breast cancer cells (MDA-MB-157). The Chromatin immunoprecipitation (ChIP) assay was done to find a potential binding site of p53 on CHAC1 promoter. Further, in vitro study was done to elucidate a potential role of CHAC1 in cell proliferation. Results: The IHC staining analysis revealed that there was no significant difference in the CHAC1 expression between high and low p53 staining tissue cores as high CHAC1 staining was observed in both cases. Further, in vitro analysis reinforced the findings of CHAC1 in TMAs as cell line studies showed that both forms of p53 had enhancing effect on CHAC1 levels. Upon induction of WT p53 and/or ectopic transfection of WT and mutant p53, we observed that CHAC1 levels were upregulated in a dose dependent manner. ChIP assay revealed that while WT p53 occupies the promoter region of the CHAC1 gene to induce its expression, mutant p53 may be inducing CHAC1 expression through an unknown mediator. These observations provide the basis that CHAC1 may be a novel transcriptional target of p53. Ectopic transfection of WT and mutant 53 showed opposite effect on cellular proliferation as evident from BrdU cellular proliferation assay in MDA-MB-157 cells. Further, we showed that CHAC1 enhanced breast cancer cell proliferation possibly via upregulation of PI3K/Akt/mTOR pathway and may likely be a player in angiogenesis as shown by in vitro angiogenesis assay. Conclusion: Taken together, the data from present study suggests that the effect of CHAC1 on cellular proliferation was associated with the status of p53 in breast cancer cells. The results suggest a plausible explanation for the differential behavior of CHAC1 in tumorigenesis. Citation Format: Vikrant Mehta, Harish Chander. Differential exploitation of Cation Transport Regulator homolog 1 (CHAC1) by Wild-type and mutant p53 [abstract]. In: Proceedings of the 2023 San Antonio Breast Cancer Symposium; 2023 Dec 5-9; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2024;84(9 Suppl):Abstract nr PO5-24-08.