Sari Kiliçaslan Seda Mehtap, Ayrim Aysun, Apaydin Elif, Incesu Zerrin
Anadolu University, Faculty of Education, Department of Primary Education, Eskişehir, Turkey.
Eskişehir Osmangazi University, Institute of Life Sciences, Eskişehir, Turkey.
Turk J Pharm Sci. 2018 Apr;15(1):50-56. doi: 10.4274/tjps.35220. Epub 2018 Apr 2.
To investigate the effects of intracellular calcium (Ca) mobilization, β-catenin and Akt signal pathways after the binding of metastatic ovarian cells to fibronectin.
The expression levels of α4β1 and αvβ6 integrin were determined using α4, β1, αv, and β6 antibodies using flow cytometry on PEO-1 cells. The effect of [Ca]i on cell adhesion capacity was investigated using RTCA after stimulating PEO-1 cells using thapsigargin and tunicamycin. The binding rate of PEO-1 cells to fibronectin was also investigated in the presence of either different concentrations of cardamonin, which inhibits the accumulation of β-catenin, or different concentrations of FPA 124, which is a specific inhibitor for the PKB/Akt signal pathway, using RTCA.
RTCA analysis results showed that increasing [Ca]i through leakage of the calcium pool was strongly effective on PEO-1 cell binding to fibronectin. Extracellular calcium influx also reduced the binding of PEO-1 cells. Cell binding to fibronectin was also inhibited with a ratio of 64% in the presence of 100 µM cardamonin compared with untreated control cells. Finally, it was found that PKB/Akt inhibition with 15 µM FPA 124 decreased the binding of PEO-1 cells to fibronectin with a ratio of 88% compared with untreated control cells.
PEO-1 cell binding to fibronectin via integrins could be related to intracellular Ca mobilization and Akt signaling.
研究转移性卵巢癌细胞与纤连蛋白结合后细胞内钙动员、β-连环蛋白和Akt信号通路的作用。
使用α4、β1、αv和β6抗体，通过流式细胞术测定PEO-1细胞上α4β1和αvβ6整合素的表达水平。在使用毒胡萝卜素和衣霉素刺激PEO-1细胞后，使用实时无标记细胞分析技术（RTCA）研究细胞内钙浓度对细胞黏附能力的影响。还使用RTCA研究了在存在不同浓度的抑制β-连环蛋白积累的小豆蔻明或不同浓度的PKB/Akt信号通路特异性抑制剂FPA 124的情况下，PEO-1细胞与纤连蛋白的结合率。
RTCA分析结果表明，通过钙库渗漏增加细胞内钙浓度对PEO-1细胞与纤连蛋白的结合有显著影响。细胞外钙内流也降低了PEO-1细胞的结合。与未处理的对照细胞相比，在存在100μM小豆蔻明的情况下，细胞与纤连蛋白的结合也受到抑制，抑制率为64%。最后发现，与未处理的对照细胞相比，用15μM FPA 124抑制PKB/Akt可使PEO-1细胞与纤连蛋白的结合率降低88%。
PEO-1细胞通过整合素与纤连蛋白的结合可能与细胞内钙动员和Akt信号传导有关。
