Impact of TLR5 and NLRC4 activation on proliferation of different breast cancer cell lines
10.3969/j.issn.1000-484X.2017.06.013
- VernacularTitle:激活TLR5和NLRC4通路的重组鞭毛素蛋白对不同乳腺癌细胞系增殖的抑制作用
- Author:
Wei LI
;
Zhaozhen ZHUO
;
Ronghui LI
;
Jun YUAN
- Keywords:
Flagellin;
TLR5;
NLRC4;
Breast cancer cell
- From:
Chinese Journal of Immunology
2017;33(6):869-872
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To explore the impact of TLR5 and NLRC4 activation on the proliferation of different breast cancer cell lines,MCF-7 and MDA-MB-23 i.Methods:Induction,expression,purification and identification of recombiant flagellin,including FliC (activating both TLR5 and NLRC4),FliC△90-97 (unable to activate TLR5),FliC-L3A (unable to activate NLRC4),FliC△90-97:L3A (unable to activate both TLR5 and NLRC4).Using different concentration of recombinant flagellin to stimulate MCF-7 and MDA-MB-231 cell lines,72 h later,the proliferation of tumor cells were detected with CCK8.We also used soft AGAR forming experiments to detect the inhibition ratio of recombinant flagellin on breast cancer cell lines.Briefly,1 000 cells were plated in the 6-well plate,then stimulated with 1 μg/ml recombinant flagellin,14 days later,the number of cloning were counted after crystal violet staining.Results:After stimulation with four recombinant flagellins at the concentration of 0.1 μ,g/ml,the inhibition ratio on MCF-7 reached 30%,and FliC△90-97 were dose-dependent on the inhibition of MCF-7 proliferation.At the concentration of 1 μg/ml,FliC-L3A which only activated TLR5 showed stronger inhibition ratio than FliC.FliC△90-97:L3A which did not activate both TLR5 and NLRC4 also inhibited the proliferation of MCF-7.After adding transfection reagent,four recombinant flagellins showed inhibition effect on MDA-MB-231.Conclusion:Flagellin can inhibit the proliferation of MCF-7 and MDA-MB-231,and the mechanism of inhibition on the proliferation were not TLR5 and NLRC4 pathway dependent.There might exist new mechanisms to explain this phenomenon.