Human lung carcinoma cell A549 express vascular endothelial grouth factor-C induced by hyaluronan / 医学研究生学报

Objective:To investigate the expression of VEGF-C in human lung carcinoma cell A549 induced by hyaluronan(HA) in vitro. Methods:A549 cells cultured in vitro were divided into three groups: control group(C): cultured with free serum medium(FSM);HA treatment group 1(HA1) and group 2(HA2): cultured with FSM containing different concentration of HA(HA1 group: 10?g/ml,HA2 group: 20?g/ml).After 48 hours,VEGF-C expression were examined by reverse transcription-polymerase chain reaction(RT-PCR)? Western blot and immunofluorescence staining. Results:Group C: both VEGF-C mRNA and protein expression were weak positive.HA1 and HA2 groups: both VEGF-C mRNA and protein expression were strong positive and increased significantly in a HA dose-depended way(P

Expression and significance of integrin-linked kinase in the process of human renal interstitial fibrosis / 第三军医大学学报

Objective To investigate the association of ILK expression in human renal interstitium and the pathological injury of renal interstitium in the patients with chronic nephropathy.Methods The expressions of ILK,TGF?1,E-cadherin and fibronectin(FN) in renal tissue were investigated with immunohistochemistry and computer image analysis system in 49 patients with chronic kidney disease.According to the score of the renal interstitial pathologic injury,49 patients were divided into four groups: 4 in no injury group(the normal control group),17 in slight injury group,16 in mild injury group,12 in severe injury group.Results ILK was hardly detected in normal control group,and ILK was observed in tubulointerstitial area in 3 injury groups.The expression of ILK was prominent in tubular epithelial cells,next in the damaged tubulointerstitial area.A significant positive correlation was found between the expression of ILK in renal interstitial tissue and the degree of renal tubulointerstitial pathological injury(r=0.736),as well as that of TGF?1(P

Lowly expression of miR-223 in CXCR4 positive cells from Lewis lung carcinoma cell line and its target gene prediction / 第三军医大学学报

Objective To analyze the differential expressions of miR-146a,miR-206,miR-223 and let-7c-1,such as cell differentiation-related miRNAs,in CXCR4-positive and CXCR4-negative subsets of the Lewis lung cancer cell lines(LLC).Methods CXCR4-positive and CXCR4-negative subsets were isolated from LLC by immunomagnetic beads sorting,and then their total cellular RNA were extracted by Trizol,expression of 4 miRNAs were detected by real-time fluorescence quantitative PCR(TaqMan probe),and potential target genes of miRNA whose differential expression was the most significant were predicted.Immunohistochemistry was carried out to confirm differential expression of the key molecule of certain research value within CXCR4-positive and CXCR4-negative subsets growing tumor tissue,and a BLAST search was performed to identify homologies of its 3′UTR.Results Compared to CXCR4-negative subsets,the expression of 4 miRNAs were lower in CXCR4-positive subsets,and expression of miR-223 had the most significant difference(Fold change=8.26).By softwares forecasting,miR-223 had potential target sites of IGF1R,IGFBP5,Pik3cb,ELK-1 and E2F1 mRNA,such as key molecular of IGF1R signaling pathway.The expression of IGF1R of CXCR4-positive subsets growing tumor tissue was significantly higher than that of CXCR4-negative subsets.Conclusion miR-223 is lowly expressed in CXCR4 positive cells from Lewis lung carcinoma cell lines.Position 238～244 nt and 688～695 nt in target sequences of 3′UTR of IGF1R mRNA was highly homologous by screening.Close correlation is found between miR-223 and IGF1R signaling pathway.The mechanisms underlying this biologically important finding need to be further explored.

Cultivation,identification and sorting of bronchoalveolar stem cells derived from mouse lung / 第三军医大学学报

Objective To cultivate,identify and sort bronchoalveolar stem cells(BASC)derived from normal adult mouse lung.Methods After enzymatic digestion of lung tissue with dispase and collagenase in combination,the Sca-1+ cells were isolated from the obtained pulmonary cells by magnetic cell sorting.These Sca-1+ cells were cultured in dishes coated with collagen and mouse fibroblast cell line Swiss-3T3 under a serum-free culture system for BASC,which were identified by the dual-color immunofluorescent staining clara cell specific antigen(CCA)and surfactant protein C(SP-C).Finally,these pure BASC were isolated by the flow cytometry.Results One lung of normal adult mouse could yield(1.6-1.8)?107 nucleated cells in this enzyme digestion procedure.The percentage of Sca-1+ cells we sorted from lung tissue was much higher than the unsorted [(87.3?5.9)% and(9.6?1.8)%,P

Highly tumorigenic and metastatic potential of CXCR4 positive cells from Lewis lung carcinoma cell line / 第三军医大学学报

0.05).Tumor formation was found in subgroups of positive CXCR4 cells(7 ? 103 ).Negative and positive CXCR cells were inoculated into 3 mice respectively at the concentration of 5 ? 105 and 2 ? 104 cells.No metastasis occurred in the former group.However,lung metastasis was found in 2 mice and ear metastasis was observed in 1mouse of the latter group.Conclusion Subgroups of positive CXCR4 cells in LLC are characterized by certain properties of cancer metastatic stem cells and have the ability to renew themselves and metabolize.

Effects of HGF on the transdifferentiation of cultured human renal epithelial cells / 解放军医学杂志

Objective To examine the possible role of hepatocyte growth factor (HGF) in transdifferentiation induced by TGF ? 1 in cultured human renal tubular epithelial cell lines(HKC). Methods The HKC cells were divided into negative control, positive control, HGF group, HGF inhibition group, HGF delayed administration group. Then the expression of E cadherin and alpha smooth muscle actin(? SMA)of HKC cells were assessed by indirect immunofluorescence ,the expression of ? SMA mRNA of HKC cells were assessed by reverse transcriptase ploymerase chain reaction(RT PCR). Results HKC cells treated with TGF ? 1 present a clear fibroblast like morphology, loss of epithelial marker E cadehrin and de novo expression of ? SMA. Simultaneous incubation of HGF or delayed administration of HGF abrogated the ? SMA expression and E cadherin depression triggered by TGF ? 1 in HKC cells in a dose dependent manner. HGF also blocked morphologic transformation of tubular epithelial cells. Conclusion TGF ? 1 is a key mediator that regulates the transdifferentiation of tubular epithelial cells into ? SMA (+) myofibroblast, HGF can inhibit transdifferentiation of HKC cells induced by TGF ? 1 , Delayed administration of HGF effectively suppresses, and partially reverses myofibroblastic transition of tubular epithelial cells