Browsing by Author "Surer, Seniz Inanc"

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Aconitine Impedes Cell Motility in Mda-Mb Breast Cancer Cells
(Dokuz Eylul Univ Inst Health Sciences, 2024) Keles, Didem; Sipahi, Murat; Surer, Seniz Inanc; Oktay, Gulgun
Purpose: Aconitine, a potent alkaloid from Aconitum plants, has shown promising anticancer properties. The aim of the study is to investigate the effects of aconitine on lateral migration, and matrix metalloproteinase (MMP) activity in MDA-MB-231 triple-negative breast cancer cells. Material and Methods: A WST-1 viability assay was conducted to determine the effect of aconitine on the viability of MDA-MB-231 cells. Following treatment with non-cytotoxic doses of aconitine, lateral migration was evaluated through wound healing assays. Additionally, gelatin zymography was conducted to analyze MMP-2 and MMP-9 activity and secretion levels. Results: Aconitine concentrations up to 200 mu M did not significantly affect cell viability for up to 72 hours, whereas higher doses (400-600 mu M) reduced viability in a time-dependent manner. Aconitine at 200 mu M showed a trend towards decreased lateral motility, with a significant reduction at 9 hours post-treatment. Gelatin zymography revealed no alterations in MMP-2 and MMP-9 activity or secretion levels following aconitine treatment. Conclusion: Aconitine demonstrates limited efficacy in modulating the migratory capacity of MDA-MB231 cells and does not affect gelatinase activity. Further investigation into underlying mechanisms is necessary, potentially leading to novel therapeutic strategies for triple-negative breast cancer.
A Simple and Efficient "Cell in Situ Collagen Zymography" Technique To Evaluate Cellular Collagenase Activities in Thyroid Cancer Cell Lines
(Springer, 2025) Savas, Ege Gokce; Surer, Seniz Inanc; Sipahi, Murat; Keles, Didem; Oktay, Gulgun
BackgroundCollagenases, a subgroup of matrix metalloproteinases (MMPs), play crucial roles in local invasion and metastasis in cancer. While substrate zymography and in situ zymography are commonly used to analyze the collagenases, traditional techniques have limitations in determining their local activities in vitro.ObjectivesWe aimed to develop a new "cell in situ collagen zymography" technique to enhance the efficiency of studying local collagenase activities in vitro.MethodsWe utilized human thyroid cancer cell lines (8505 C, B-CPAP, FTC-133) and normal follicular thyroid cell line (Nhty-ori-3-1). We compared collagenase levels across these cell lines and selected 8505 C as a model due to its highest collagenase activity. We optimized factors including (i) fixation method (methanol, ethanol and zinc), (ii) dye-quenched (DQ) collagen concentration and (iii) collagen gel configuration. For gel configuration, cells were seeded under, on the top of, or between (sandwich) collagen gel layers. As controls, enzymatic activity was suppressed in the presence of EDTA, piroxicam and matrix metalloproteinase 8 inhibitor I. The optimized method was also applied to BCPAP, FTC-133, and Nthy-ori-3-1.ResultsOur optimization process revealed that that the best visualization of collagenase activity in 8505 C was provided by the "sandwich model" of gel, containing 25 mu g/mL of DQ-collagen with 100% cold methanol fixation. We confirmed the optimized method's applicability in other thyroid cell lines. The use of inhibitors validated the specificity of the fluorescent signal to MMP activity.ConclusionThe innovative "cell in situ collagen zymography" technique offers an efficient, cost-effective, and rapid method for analyzing local collagenase activities in vitro.