Abstract 3840: Rap1 GTPase activating protein (Rap1GAP) is downregulated in DCIS and invasive ductal carcinoma

Abstract

Introduction: Rap1 and Rap2, both small GTPases of the Ras family, serve as crucial regulators in cell polarity, adhesion, migration, and lumen formation via crosstalk with cytoskeletal proteins. Rap1 GTPase activating protein (Rap1GAP), which negatively regulates Rap1 and Rap2 activity, has found to be downregulated in multiple cancers, including oropharyngeal squamous cell carcinoma, melanoma, papillary and follicular thyroid carcinoma, and colorectal carcinoma but has not yet been studied in breast cancer. In order to determine if Rap1GAP downregulation plays a role in breast cancer formation, we decided to expand our prior work to study its expression in breast cancer tissue. Methods: Twenty-one formalin-fixed paraffin-embedded human tissue blocks were selected on the basis that they contained normal breast tissue and either adjacent breast ductal carcinoma in situ (DCIS) or invasive ductal carcinoma. Using immunohistochemistry, we assessed the expression of Rap1GAP. We also assayed two human breast cancer cell lines with different levels of invasiveness, MCF7 (non-invasive) and MDA-MB-231 (invasive), to determine different levels of Rap1GAP expression and subsequent changes in Rap1 and Rap2 expression and activity. Results: Rap1GAP expression in both DCIS and invasive ductal carcinoma were found to be decreased compared to adjacent normal breast tissue. However, Rap1GAP expression was not statistically different between DCIS and invasive carcinoma lesions. Rap1GAP was found to be expressed highly in MCF7 but not in MDA-MB-231 cells. Moreover, only MDA-MB-231 cells exhibited high basal Rap2 activity. Activated Rap1 (RAP1-GTP) was not detected in either line. Conclusion: Our findings suggest that loss of Rap1GAP occurs as an early event in breast cancer tumorigenesis with a significant decrease in the transition from normal to DCIS, with a possible further decrement in the DCIS to invasive transition. Interestingly, while Rap1 and Rap2 were expressed in non-invasive and invasive cell lines, loss of Rap1GAP in the invasive cell line was associated with increase in active Rap2, but not active Rap1. These findings suggest that as in other cancers, loss of Rap1GAP may contribute to the invasive phenotype and thus further investigation is warranted. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 3840. doi:10.1158/1538-7445.AM2011-3840

Downregulation of Rap1Gap: A Switch from DCIS to Invasive Breast Carcinoma via ERK/MAPK Activation

“…In the two other DCIS lines (SUM 225 and SUM 102), Rap1Gap levels were strikingly different, although prior transcriptomic analysis identified the upregulation of Rap1Gap mRNA in both [31] . SUM 225 cells, a HER2 overexpressing line that exhibits luminal markers and was derived from a chest wall recurrence in a patient previously diagnosed with DCIS [32] , abundantly expressed Rap1Gap to a level similar to that found in MCF-7 cells, which have previously been characterized to have high expression [33] . In contrast, SUM 102 cells, which are of basal type and were derived from a DCIS with microinvasion [34] , [35] , expressed very low levels of Rap1Gap.…”