The aimof this study was to screen cytotoxicity biomarkers of nickel ions(Ni^(2+))using transcriptomic and proteomic approaches combined with molecular biology validation.First,the MTT method was used to evaluate cyto...The aimof this study was to screen cytotoxicity biomarkers of nickel ions(Ni^(2+))using transcriptomic and proteomic approaches combined with molecular biology validation.First,the MTT method was used to evaluate cytotoxicity in L929 cells treated with Ni^(2+)at different concentrations.Ni^(2+)at both 100μM and 200 lMaffected cell proliferation.Then,transcriptomic and proteomic technology was used to study the effects of Ni^(2+)on the expression of genes/proteins in cells.It was found that 1490,789,652 and 729 genes(12,24,48 and 72 h,respectively)and 177,2191 and 2095 proteins(12,24 and 48 h,respectively)were differentially expressed after treatment with 100μM Ni^(2+).In total,1403,963,916 and 1230 genes(12,24,48 and 72 h,respectively)and 83,1681 and 2398 proteins(12,24 and 48 h,respectively)were differentially expressed after treatment with 200μMNi^(2+).Then,four target gene/protein biomarkers were filtered by combined screening using gene/proteomic experimental data and biological pathway analyses.Further expression level validation of all these target biomarkers and functional validation of selected gene/protein biomarkers were carried out,and a final gene/protein biomarker(UQCRB)was identified.展开更多
The purpose of this paper is to utilize the signaling pathway polymerase chain reaction(PCR)arrays to investigate the activation of two important biological signaling pathways in endothelial cell adhesion and growth m...The purpose of this paper is to utilize the signaling pathway polymerase chain reaction(PCR)arrays to investigate the activation of two important biological signaling pathways in endothelial cell adhesion and growth mediated by adsorbed serum protein on the surface of bare and titanium nitride(TiN)-coated nickel titanium(NiTi)alloys.First,the endothelial cells were cultured on the bare and TiN-coated NiTi alloys and chitosan films as control for 4 h and 24 h,respectively.Then,the total RNA of the cells was collected and the PCR arrays were performed.After that,the differentially expressed genes in the transforming growth factor beta(TGF-b)signaling pathway and the regulation of actin cytoskeleton pathway were screened out;and the further bioinformatics analyses were performed.The results showed that both TGF-b signaling pathway and regulation of actin cytoskeleton pathway were activated in the cells after 4 h and 24 h culturing on the surface of bare and TiN-coated NiTi alloys compared to the chitosan group.The activated TGF-b signaling pathway promoted cell adhesion;the activated regulation of actin cytoskeleton pathway promoted cell adhesion,spreading,growth and motility.In addition,the activation of both pathways was much stronger in the cells cultured for 24 h versus 4 h,which indicated that cell adhesion and growth became more favorable with longer time on the surface of two NiTi alloy materials.展开更多
基金supported by the National Natural Science Foundation of China(31971254).
文摘The aimof this study was to screen cytotoxicity biomarkers of nickel ions(Ni^(2+))using transcriptomic and proteomic approaches combined with molecular biology validation.First,the MTT method was used to evaluate cytotoxicity in L929 cells treated with Ni^(2+)at different concentrations.Ni^(2+)at both 100μM and 200 lMaffected cell proliferation.Then,transcriptomic and proteomic technology was used to study the effects of Ni^(2+)on the expression of genes/proteins in cells.It was found that 1490,789,652 and 729 genes(12,24,48 and 72 h,respectively)and 177,2191 and 2095 proteins(12,24 and 48 h,respectively)were differentially expressed after treatment with 100μM Ni^(2+).In total,1403,963,916 and 1230 genes(12,24,48 and 72 h,respectively)and 83,1681 and 2398 proteins(12,24 and 48 h,respectively)were differentially expressed after treatment with 200μMNi^(2+).Then,four target gene/protein biomarkers were filtered by combined screening using gene/proteomic experimental data and biological pathway analyses.Further expression level validation of all these target biomarkers and functional validation of selected gene/protein biomarkers were carried out,and a final gene/protein biomarker(UQCRB)was identified.
基金National Natural Science Foundation of China(31271012)973 Project(No.2009CB930000)the Natural Science Foundation of Jiangsu Province(BK20150599).
文摘The purpose of this paper is to utilize the signaling pathway polymerase chain reaction(PCR)arrays to investigate the activation of two important biological signaling pathways in endothelial cell adhesion and growth mediated by adsorbed serum protein on the surface of bare and titanium nitride(TiN)-coated nickel titanium(NiTi)alloys.First,the endothelial cells were cultured on the bare and TiN-coated NiTi alloys and chitosan films as control for 4 h and 24 h,respectively.Then,the total RNA of the cells was collected and the PCR arrays were performed.After that,the differentially expressed genes in the transforming growth factor beta(TGF-b)signaling pathway and the regulation of actin cytoskeleton pathway were screened out;and the further bioinformatics analyses were performed.The results showed that both TGF-b signaling pathway and regulation of actin cytoskeleton pathway were activated in the cells after 4 h and 24 h culturing on the surface of bare and TiN-coated NiTi alloys compared to the chitosan group.The activated TGF-b signaling pathway promoted cell adhesion;the activated regulation of actin cytoskeleton pathway promoted cell adhesion,spreading,growth and motility.In addition,the activation of both pathways was much stronger in the cells cultured for 24 h versus 4 h,which indicated that cell adhesion and growth became more favorable with longer time on the surface of two NiTi alloy materials.
