RESUMO
Objective: To investigate the effects of up-regulating or silencing peroxiredoxin 4 (Prd×4) protein expression on the proliferation, apoptosis and migration of human lung adenocarcinoma A549 cells. Methods: The recombinant plasmid pcDNA3.0-HA-Prd×4 and specific siRNA targeting human Prdx 4 gene (Prd×4-siRNA) were transfected into A549 cells by liposome, respectively; while the empty vector pcDNA3.0-HA and the negative control-siRNA were used as the controls. The expression levels of Prd×4 mRNA and protein were detected by real-time fluorescent quantitative-PCR and Western blotting, respectively. The proliferation and apoptosis of A549 cells were detected by CCK-8 assay and immunofluorescence assay, respectively. The migration ability of A549 cells was detected by wound-healing test and Transwell chamber assay. Results: The expression levels of Prd×4 mRNA and protein were up-regulated in A549 cells transfected with the recombinant plasmid pcDNA3.0-HA-Prd×4 (both P < 0.05), while they were down-regulated in A549 cells transfected with Prd×4-siRNA (both P < 0.01). The proliferation ability (D 450 nm) of A549 cells with Prd×4 over-expression was higher than that in the control group at 24, 48 and 72 h (all P < 0.05), and the difference at 96 h was most obvious (P < 0.01). The D 450 nm values of A549 cells with Prdx 4 gene-silencing were decreased (all P < 0.01) as compared with the control group at 24, 48, 72 and 96 h. After treatment with cisplatin for 24 and 48 h, the number of apoptotic cells in Prd×4 over-expression group was significantly decreased as compared with the control group (P < 0.05 and P < 0.01), while it was significantly increased in Prdx 4 gene-silencing group as compared with the control group (P < 0.05 and P < 0.01). The wound healing rate in Prd×4 over-expression group was higher than that in the control group (P < 0.01), while it was lower in Prdx 4 gene-silencing group than that in the control group (P < 0.01). The number of A549 cells passing through the membrane of Transwell chamber in Prd×4 over-expression group was significantly more than that in the control group (P < 0.01), while it was less in Prdx 4 genesilencing group than that in the control group (P < 0.01). Conclusion: Prd×4 can promote the proliferation and migration, while it can inhibit the apoptosis of A549 cells. Prd×4 may be a potential target for the clinical treatment of lung adenocarcinoma.
RESUMO
Objective: To screen the differentially expressed proteins in clear cell renal cell carcinoma (ccRCC), and to investigate its clinical significance. Methods: The differentially expressed proteins in 15 cases of ccRCC and their adjacent renal tissues were screened by two-dimensional fluorescence difference in gel electrophoresis combined with mass spectrometry (2-D DIGE-MS). The expression levels of differentially expressed proteins screened by 2-D DIGE-MS in 45 cases of ccRCC and their adjacent tissues were examined by Western blotting. The expression levels of this differentially expressed proteins in 152 specimens of ccRCC and 40 specimens of adjacent tissues were detected by immunohistochemistry. The correlations of this protein expression with the clinicopathologic features and prognosis of ccRCC patients were analyzed. Results: Proteomics screening results showed that the expression level of peroxiredoxin 4 (PRDX4) was significantly different in ccRCC and its adjacent tissues; compared with the adjacent tissues, PRDX4 expression was significantly down-regulated in ccRCC tissues (P 0.05). The patients with lower expression of PRDX4 had high grade of ccRCC (? = -0.211, P = 0.009) and distant metastasis (? = -0.161, P = 0.048). The 5-year survival rates of patients with positive and negative expression of PRDX4 were 75.3% and 62.7% (P = 0.862), respectively. Conclusion: PRDX4, as a differentially expressed protein in ccRCC, may be involved in the formation and development of renal cancer, and its down-regulation is correlated with the increased malignancy of ccRCC.