ABSTRACT
ObjectiveTo investigate the expression of growth factor receptor-bound protein-2 (Grb2), matrix metalloproteinase-3 (MMP-3), and MMP-9 in cholangiocarcinoma and its significance. MethodsThe expression of Grb2, MMP-3, and MMP-9 in cholangiocarcinoma tissues of 47 cases and normal tissues was measured using immunohistochemistry, and the correlations of Grb2 expression with clinical pathology and MMP-3 and MMP-9 expression were analyzed. Comparison of continuous data was made using t test, and the correlation of Grb2 expression with MMP-3 and MMP-9 expression was analyzed using the multivariate linear regression model. ResultsThe expression of Grb2 in cholangiocarcinoma tissues was significantly higher than that in normal bile duct tissues (t=5935, P＜0.001); the expression of Grb2 in cholangiocarcinoma tissues and normal bile duct tissues showed no significant correlation with age, sex, and differentiation level; the expression of Grb2 in cholangiocarcinoma tissues with lymph node or distant metastasis was significantly higher than that in cholangiocarcinoma tissues without metastasis (t=3.882, P=0.003). The expression of Grb2 was positively correlated with the expression of MMP-3 and MMP-9 (r2=0.3667, P=0.018; r2=0.5133, P=0.007). ConclusionThe expression of Grb2 in cholangiocarcinoma tissues is higher than that in normal bile duct tissues, and it is closely related to the invasion and metastasis of carcinoma. Further study shows that the expression of Grb2 is positively correlated with the expression of MMP-3 and MMP-9.
ABSTRACT
Objective: To testify the location of recombinant adenovirus Ad5/F35-HF2S in chronic myeloid leukemia K562 cells and its interaction with the fusion protein Bcr-Abl. Methods: The recombinant adenovirus Ad5/F35-HF2S was infected into K562 cells, then the expression of fusion protein HF2S in K562 cells was assessed by RT-PCR (reverse transcription-PCR) and Western blotting. The location of HF2S protein in K562 cells was demonstrated by immunofluorescence and Western blotting assay. The immunofluorescence was performed to verify the co-localization of HF2S and Bcr-Abl. The co-immunoprecipitation assay was employed to detect the interaction between HF2S and Bcr-Abl. Results: The leukemia cell line K562 was effectively infected by the adenovirus Ad5/F35-HF2S. The fusion protein HF2S and Bcr-Abl were co-located and interacted with each other in the cytoplasm. Conclusion: HF2S can be successfully expressed and interacted with Bcr-Abl in the cytoplasm of K562 cells. These results provide the basement of the targeted therapy for chronic myeloid leukemia. Copyright © 2013 by TUMOR.
ABSTRACT
Objective To explore the role of growth factor receptor-bound 2 (GRB2) in homo sapiens eukaryotic translation elongation factor 1 alpha 2 (EEF1A2) promoting the carcinogenesis and development of pancreatic cancer.Methods The human pancreatic cancer cell line SW1990 with low expression of EEF1A2,was transfected by Ad5/F35-EEF1A2 plasmid.The expression of EEF1A2 in human pancreatic cancer cell lines BxPC-3 cells with high expression of EEF1A2,was down-regulated by small interfering RNA (siRNA) interference.The expressions of GRB2 in SW1990 and BxPC-3 cells were detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot before and after interference.The pancreatic cancer BxPC-3 cells were interfered by specific and efficient chemical synthesized siRNA fragment.The changes of BxPC-3 cell proliferation and migration were observed by methyl thiazoly tetrazolium (MTT) and Transwell assay before and after interference.The data were analyzed by one-way analysis of variance.Results After human pancreatic cancer cell line SW1990 was transfected with Ad5/F35-EEF1A2 plasmid,the expression of GRB2 at mRNA and protein level both increased significantly (F=5.67,6.39,both P＜0.05).After the expression of EEF1A2 was inhibited by EEF1A2-siRNA,the expression of GRB2 at mRNA and protein level both decreased significantly (F=6.59,4.69,both P＜0.05).After BxPC-3 cells was transfected with siRNA-GRB2 for 72 hours,the results of MTT assay indicated that the absorbance value was 1.22±0.14,compared with negative control group (1.42±0.15) and blank control group (1.39 ± 0.15) the difference was statistically significant (F =6.63,P＜ 0.05).The results of transwell assay showed that the migration ability of cells in siRNA-GRB2 group decreased.After 24 hours,the number of migrated cells was 24.10±4.25,compared with negative control group (54.72±5.24) and blank control group (51.26 ± 6.23) the difference was statistically significant (F=55.00,P＜ 0.05).Conclusion GRB2 is key molecule in EEF1A2 promoting the progenesis and development of pancreatic cancer.
ABSTRACT
Objective To analyze the influence of peptidimer-c on the gene expression profiling of K562 cells and investigate the mechanism of peptidimer-c inducing the apoptosis and inhibiting proliferation of K562cells.Methods Trypan blue staining technique was performed for counting the number of living K562 cells treated with peptidimer-c.The ultrastructure changes of K562 cells treated with peptidimer-c was observed under transmission electron microscope.The Human U133 Plus 3.0 gene chips were used to detect the differentially expressed genes of K562 cells treated with peptidimer-c.Reverse transcription PCR was conducted to confirm some genes identified by gene chips.Results Peptidimer-c could induce the apoptosis and inhibit the proliferation of K562 cells.Peptidimer-c caused widely changes of the gene expression profiles of K562 cells.The chip data suggested that there were 529 differentially expressed genes,of which 455 genes were up-regulated and 74 genes were down-regulated.The relevant apoptotic genes were down-regulated markedly,including JUN,AXUD1,TNFRSF10B,etc.Fifteen of the differentially expressed genes were detected by RT-PCR,which was consistent with the chip data.Conclusion Peptidimer-c may induce aooptosis of K562 cells by activating the TNF/TNFR family and the JUN family.