ABSTRACT
Objective To investigate the effect of over-expressed macrophage migration inhibitory factor (MIF) on epithelial-mesenchymal transition (EMT) in human cervical carcinoma SiHa cells.Methods Recombinant eukaryotic expression plasmid liposome enhanced transfection of green fluorescent protein gene (pEGFP-N1)-MIF was constructed and then transfected into human cervical cancer SiHa cells.Experimental cells were classified into three groups (SiHa-pEGFP-N1-MIF,SiHa-pEGFP-N1,and SiHa).Western blot was used to detect the expression of MIF protein,and the expressions of EMT-related markers such as E-cadherin and vimentin in SiHa cells were determined before and after transfection.Results The eukaryotic expression vector pEGFP-N1-MIF significantly increased the expression of MIF protein in SiHa cells (P ＜ 0.05),and after overexpression of MIF gene in SiHa cells,the expression of E-cadherin protein in SiHa-pEGFP-N1-MIF group was significantly lower than that in control groups (P ＜0.05),while the expression of vimentin in SiHa-pEGFP-N1-MIF group was significantly higher than that in control groups (P ＜ 0.05).Conclusions Overexpression of MIF in cervical cancer SiHa cells can promote the EMT occurrence.
ABSTRACT
Objective To study the effect of calcium on human peritoneal mesothelial cells (HPMCs). Methods Proliferation abilities of HPMCs were assessed by tetrazolium salt colorimetry assay (MTT assay) and the levels of LDH in the supernatant were detected in all groups to evaluate the damage of HPMCs. The expression of TNF-α in cytoplasm was detected by immunohistochemistry. Results Calcium enhanced proliferation of HPMCs in a time-dependent manor(P＜0.01), especially calcium with 1.25 mmol/L(0.5098±0.016,0.6763±0.048) and 1.0 mmol/L(0.4853±0.016,0.6678±0.076). Calcium with different concentrations significantly increased the levels of LDH in HPMCs in a time-dependent manor, while the effect of calcium with 1.25 mmol/L was lowest(17.78±1.18,23.60±1.39,P＜0.01). Calcium with 2.0 mmol/L[(42.61±3.29)%] and 1.75 mmol/L[(33.32±1.88)%] significantly up-regulated the expression of TNF-α(P＜0.01) . Conclusions High calcium damaged HPMCs and up-regulated the expression of TNF-αby HPMCs, while physical calcium (1.25 mmol/L)can protect peritoneum and prevent peritoneal fibrosis.
