Secreted proteins are important sources for early detection and diagnosis of disease, and as such have received considerable attention. The extraction of low concentration proteins from large volumes of culture media,...Secreted proteins are important sources for early detection and diagnosis of disease, and as such have received considerable attention. The extraction of low concentration proteins from large volumes of culture media, which are rich in salts and other compounds that interfere with most proteomics techniques, presents a problem for secretome studies. Ultrafiltration, precipitation, and dialysis are three major extraction methods that can be used to overcome this problem. The present study for the first time, compared the merits and shortcomings of these three methods, without bias. Centrifugal ultrafiltration provided the best extraction efficiency, and precipitation provided the highest number of identifiable proteins. The three methods yielded closely related, but different, information on the secretome; thus, they should be considered complementary or, at least, supplementary methods. Three hundred and sixty unique proteins were identified, including 211 potential secreted proteins. Compared with previous studies, this study also identified 42 new secreted proteins. The present study not only offers a reference for the selection of secretome extraction methods, but also expands the secretome database for the investigation of hepatocellular carcinoma.展开更多
Galectin-3(gal-3)and its ligands have been implicated in cell transformation and cancer metastasis.Gal-3 protein has been found in uterine epithelial cells adjacent to implanting blastocysts in different cell types.In...Galectin-3(gal-3)and its ligands have been implicated in cell transformation and cancer metastasis.Gal-3 protein has been found in uterine epithelial cells adjacent to implanting blastocysts in different cell types.In order to investigate the role of gal-3 in the establishment of human endometrial receptivity,the expression of gal-3 in human endometrial cell line RL95-2 was silenced by RNA inter-ference technology using gal-3 specific small RNA.The expression of gal-3 was detected by the reverse transcriptase-polymerase chain reaction and Western blot analysis.After the suppression of gal-3,cell cycle changes and the expres-sion of integrin b1 were detected by flow cytometry.The adhesive ability of RL95-2 cells was analyzed by the adhesion test.Gal-3 siRNA transfection efficiency reached 70%–90%.The expression of gal-3 mRNA and protein in RL95-2 cells was strongly inhibited by 70%–90%after RNA interference.Inhibition of gal-3 expression decreased S-phase but increased G1 phase cells.Integrin b1 expression was down-regulated,and the adhesive ability of RL95-2 cells to fibronectin(FN)was significantly reduced.Gal-3 may be involved in the establishment of endometrial receptivity by regulating the proliferation and adhesion of endometrial cells.The influence on adhesion may be related with the integrin modulation.展开更多
基金supported by the National Natural Science Foundation of China (Grant No. 209750240)the National Basic Research Program of China (Grant No. 2010CB912700)
文摘Secreted proteins are important sources for early detection and diagnosis of disease, and as such have received considerable attention. The extraction of low concentration proteins from large volumes of culture media, which are rich in salts and other compounds that interfere with most proteomics techniques, presents a problem for secretome studies. Ultrafiltration, precipitation, and dialysis are three major extraction methods that can be used to overcome this problem. The present study for the first time, compared the merits and shortcomings of these three methods, without bias. Centrifugal ultrafiltration provided the best extraction efficiency, and precipitation provided the highest number of identifiable proteins. The three methods yielded closely related, but different, information on the secretome; thus, they should be considered complementary or, at least, supplementary methods. Three hundred and sixty unique proteins were identified, including 211 potential secreted proteins. Compared with previous studies, this study also identified 42 new secreted proteins. The present study not only offers a reference for the selection of secretome extraction methods, but also expands the secretome database for the investigation of hepatocellular carcinoma.
基金supported by the National Natural Science Foundation of China(Grant No.30271354).
文摘Galectin-3(gal-3)and its ligands have been implicated in cell transformation and cancer metastasis.Gal-3 protein has been found in uterine epithelial cells adjacent to implanting blastocysts in different cell types.In order to investigate the role of gal-3 in the establishment of human endometrial receptivity,the expression of gal-3 in human endometrial cell line RL95-2 was silenced by RNA inter-ference technology using gal-3 specific small RNA.The expression of gal-3 was detected by the reverse transcriptase-polymerase chain reaction and Western blot analysis.After the suppression of gal-3,cell cycle changes and the expres-sion of integrin b1 were detected by flow cytometry.The adhesive ability of RL95-2 cells was analyzed by the adhesion test.Gal-3 siRNA transfection efficiency reached 70%–90%.The expression of gal-3 mRNA and protein in RL95-2 cells was strongly inhibited by 70%–90%after RNA interference.Inhibition of gal-3 expression decreased S-phase but increased G1 phase cells.Integrin b1 expression was down-regulated,and the adhesive ability of RL95-2 cells to fibronectin(FN)was significantly reduced.Gal-3 may be involved in the establishment of endometrial receptivity by regulating the proliferation and adhesion of endometrial cells.The influence on adhesion may be related with the integrin modulation.
