1.Effect of heat shock protein 47 on epithelial-mesenchymal transdifferentiation induced by transforming growth factor β1 in the renal tubular epithelial cells
Huixin BI ; Ruihong LIU ; Lin SUN ; Hong LIU ; Shuguang YUAN ; Guxiang HUANG ; Nanlan CHEN
Chinese Journal of Nephrology 2013;29(10):775-781
Objective To detect the expression of heat shock protein 47(HSP47) in renal proximal epithelial cell lines (HK-2) and to investigate the role of HSP47 in the progress of transforming growth factor β1 (TGF-β1) induced epithelial-mesenchymal transdifferentiation (EMT) in HK-2 cells.Methods HK-2 cells were exposed to TGF-β1 (0,2.5,5,10 μg/L) for different time (0,12,24,48 h).The expression of HSP47 was examined by Western blotting.Then HK-2 cells were exposed to 10 μg/L TGF-β1,the expressions of vimentin,zona occludens-1 (ZO-1) were examined by Western blotting and real-time PCR.Furthermore,the expressions of p-Smad3 and Smad3 were examined by Western blotting.HK-2 cells were transfected with HSP47 siRNA and siRNA negative control before exposing to TGF-β1.Then the expressions of vimentin,ZO-1 were detected by Western blotting and real-time PCR,meanwhile Western blotting for HSP47,p-Smad3 and Smad3.Results Stimulating HK-2 with TGF-β1 resulted in a significant increased expression of HSP47 in time-and concentration-dependent manner (P < 0.05).Meanwhile,TGF-β1 up-regulated the protein and mRNA expression of vimentin (P < 0.05),and down-regulated the protein and mRNA expression of ZO-1 (P < 0.05),all in time-dependent manner.Stimulating HK-2 with TGF-β1 resulted in phosphorylation of Smad3,which was peaked at 30 min,slightly decreased at 1 h,and then increased again between 24 and 48 h (P < 0.05).Compared to the TGF-β1 group,inhibition of HSP4.7 expression in HK-2up-regulated the protein and mRNA expression of ZO-1,down-regulated the protein and mRNA expression of vimentin (P < 0.05) and down-regulated the ratio of p-Smad3/Smad3.HSP47 siRNA negative control had no significant effect on the expressions of ZO-1,vimentin and p-Smad3/Smad3 (P > 0.05).Conclusion HSP47 can promote the EMT of renal tubular epithelial cell which is possibly via the TGF-β1-Smad3 pathway.
2.Relationship between the expression ofCEACAM1, CD34 and the metastasis of gastric cancer
Qi PAN ; Yan CHEN ; Pei XIANG ; Yijiao HUANG ; Nanlan XIE ; Ye ZHANG
Basic & Clinical Medicine 2017;37(5):705-708
Objective To investigate the prognostic impact of carcinoembryonic antigen-related cell adhesion molecule 1 (CEACAM1) and CD34 on invasion and metastasis of gastric cancer.Methods CEACAM1 and CD34 expressions were detected by immunhistochemistry in 90 cases of gastric cancer and 30 cases of normal gastric mucosa.Then the relationship among CEACAM1, MVD marked by CD34 antibody and gastric cancer patients` clinical pathological features and prognosis were analyzed.Results The positive rates of CEACAM1 and CD34 protein in gastric cancer were significantly higher than those in normal gastric mucosa (P<0.05).The positive rate of CEACAM1 was closely related to histological differentiation, tumor invasion, nodal metastasis and TNM stage.MVD marked by CD34 was related with tumor size, histological differentiation, tumor invasion, nodal metastasis and TNM stage(P<0.05).Conclusions In gastric cancer, CEACAM1 and CD34 promote its invasion and metastasis, and they are potential indicators of predicting development and prognosis.