Abstract 83: Investigating the effect of SERPINB3 and STAT3 on invasion and metastasis in cervical cancer cells

Abstract Cervical cancer is the fourth most leading cause of cancer related deaths in women worldwide. Of the women treated for cervical cancer, nearly a third experience recurrence after initial treatment with chemotherapy and radiation. After recurrence, salvage rates are low, thus better treatment options must be developed. One biomarker in cervical cancer is Squamous Cell Carcinoma Antigen 1, also known as SERPINB3. SERPINB3, a member of the serine protease inhibitor superfamily, has been found in elevated levels in patients with cervical cancer, and correlates with resistance to chemo and ionizing radiation. Signal Transducer and Activator of Transcription 3 (STAT3) activates signaling pathways that lead to tumor progression including cancer cell migration and metastasis. In this project, we investigated how SERPINB3 and STAT3 correlate to cervical cancer cell migration and metastasis. Our hypothesis is that SERPINB3 interacts with STAT3 to increase metastasis of cervical cancer cells. To test this hypothesis, we used the genetically modified SW-756 cervical cancer cell line with and without SERPINB3 and performed transient transfection to knock down the STAT3 gene using silencing RNA (siRNA). To analyze the knock down of STAT3, we used immunoblotting and quantitative polymerase chain reaction (qPCR) to determine whether the expression of the STAT3 protein and gene was silenced. To determine metastasis of the cells in vitro, we conducted a wound healing assay, in which a wound is generated in the middle of the tissue culture well, and the closure of the wound is monitored every 24 hours via microscopy. The images were analyzed using FIJI Image J Wound Healing Program. In this project, we have minimized the expression of STAT3 in our cervical cancer cell lines. We noticed that when we decreased STAT3 expression, SERPINB3 levels had also decreased. This correlates with literature suggesting that STAT3 regulates SERPINB3 expression. The wound healing assays revealed that upon knocking down STAT3 in SERPINB3 knockout cells, cell migration had severely decreased compared to SERPINB3 control cell lines knocked down with STAT3. The knockdown will be optimized and results repeated along with pharmacologic inhibition of STAT signaling to determine the mechanism of B3 in migration and metastasis phenotypes. The future direction of this project is to identify the exact SERPINB3-STAT3 interaction that leads to invasion metastasis in cervical cancer. By doing this, we hope to determine different pathways that may lead to increased invasion and metastasis to develop better treatment options in cervical cancer. Citation Format: Marlene Campos Guerrero, Darya E. Hasanova, Li-Yun Chen, Leah Sherbansky, Stephanie Markovina. Investigating the effect of SERPINB3 and STAT3 on invasion and metastasis in cervical cancer cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2025; Part 1 (Regular Abstracts); 2025 Apr 25-30; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2025;85(8_Suppl_1):Abstract nr 83.