Abstract 4877: Mutant or overexpressed SF3B1 facilitates endometrial cancer tumorigenesis
Authors: Popli Pooja , Deng Xinchao , Thaker Premal , Kommagani Ramakrishna 
Published: April 2023

Aberrant expression or mutations in core splicing factors including U2AF1, SRSF2, ZRSR2, and SF3B1 found to drive tumorigenesis of various malignant tumors e.g., myelodysplastic syndromes, chronic lymphocytic leukemia, uveal melanoma, and breast cancer. Importantly, the amplifications and mutations in SF3B1, the pioneer splicing factor, are reportedly far more common than those in other splicing factors. However, the functional relevance and underlying mechanisms of dysregulated SF3B1 in endometrial cancer are not investigated. Endometrial cancer is the fourth most common cancer in women in the United States and there will be about 65,950 new endometrial cancer cases in the current year alone with 12,550 estimated cancer deaths in the United States. In this study, we thus investigated the role of SF3B1 in endometrial adenocarcinoma. We found SF3B1 is one of the most frequently amplified or mutated splicing factors in endometrial cancer. Specifically, analysis of The Cancer Genome Atlas (TCGA) uterine pan-cancer dataset revealed that up to 9% of endometrial cancers have SF3B1 mutations or amplification. Additionally, we showed that the SF3B1 protein is overexpressed in human as well as mouse endometrial tumor samples. Further analysis found elevated SF3B1 protein in multiple endometrial cancer cell lines even in the absence of SF3B1 mutations. However, we did not observe any altered expression at the transcript level. Further, we identified a correlation between the over-expressed SF3B1 levels and the tumorigenicity/invasive potential of cancer cell lines. For instance, endometrial cancer cell lines (Ishikawa, AN3CA, KLE) with over-expressed SF3B1 levels were much more tumorigenic and invasive than those with low endogenous SF3B1 levels (normal human endometrial epithelial cells, RL-95-2 cells). Moreover, cell viability assays revealed that Ishikawa, AN3CA, and KLE cells were more susceptible to the loss of SF3B1 or treatment with the SF3B1 inhibitor Pladienolide-B (PLAD-B) than RL-95-2 cells. To further ensure that overexpressed or mutated SF3B1 drives endometrial tumorigenesis, we transiently transfected an empty vector or a vector expressing FLAG-tagged wild-type SF3B1 or vector with mutated SF3B1 construct R957Q into RL-95-2 cells which have minimal endogenous SF3B1. Analysis of proliferation assays revealed that cells overexpressing SF3B1 proliferated significantly more than cells expressing empty vector. Intriguingly, we observed that RL-95-2 cells over-expressing R957Q mutants were even more proliferative than the cells expressing wild-type SF3B1. Together, this data suggest that SF3B1 over-expression and mutation play a crucial oncogenic role in endometrial cancer, and these findings may support the development of SF3B1 inhibitors to treat this disease.

Citation Format: Pooja Popli, Xinchao Deng, Premal Thaker, Ramakrishna Kommagani. Mutant or overexpressed SF3B1 facilitates endometrial cancer tumorigenesis. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 4877.