[The inhibitory effects of siRNA expression vector on cell proliferation and expression of hnRNPB1 gene in lung cancer A549 cells].
Dan Pu, Weimin Li, Taoyou Zhou, Di Wu, Xiaobing Chen, Min Chen, Feng-ming Tang, Juan Han
Abstract
Dan Pu, Weimin Li, Taoyou Zhou, Di Wu, Xiaobing Chen, Min Chen, Feng-ming Tang, Juan Han
Abstract
OBJECTIVE: To investigate the inhibitory effects of specific siRNA on the expression of heterogeneous nuclear ribonucleoproteins (hnRNPB1) and cell proliferation in lung cancer A549 cells. METHODS: After the construction of siRNA expression vector for hnRNPB1, cultured A549 cells were tansfected with the specific siRNA vector, so RNA of A549 cells was interfered by RNA interference technique. The cells were retrieved at 1st, 4th and 6th week after transfection, cell cycles and cell apoptosis were measured with flow cytometry, the mRNAs levels of hnRNPB1, were detected by fluorescence quantity RT-PCR, the protein levels of hnRNPB1 was detected by Western blot technique. RESULTS: It was found that plasmid-derived siRNAs could inhibit hnRNPB1 mRNA and protein expression. The expressions of hnRNPB1 mRNA and protein were decreased about 46%-73%, and 77.69%-83.04% respectively. We also observed that siRNA in A549 cell suppressed the cell proliferation by increasing the amounts of G1 cells, and induced cell apoptosis. CONCLUSION: These results demonstrated the effectiveness of siRNA in lung caner cell, which indicated a possible clinical therapy method for lung cancer.
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OBJECTIVE: To investigate the inhibitory effects of specific siRNA on the expression of heterogeneous nuclear ribonucleoproteins (hnRNPB1) and cell proliferation in lung cancer A549 cells. METHODS: After the construction of siRNA expression vector for hnRNPB1, cultured A549 cells were tansfected with the specific siRNA vector, so RNA of A549 cells was interfered by RNA interference technique. The cells were retrieved at 1st, 4th and 6th week after transfection, cell cycles and cell apoptosis were measured with flow cytometry, the mRNAs levels of hnRNPB1, were detected by fluorescence quantity RT-PCR, the protein levels of hnRNPB1 was detected by Western blot technique. RESULTS: It was found that plasmid-derived siRNAs could inhibit hnRNPB1 mRNA and protein expression. The expressions of hnRNPB1 mRNA and protein were decreased about 46%-73%, and 77.69%-83.04% respectively. We also observed that siRNA in A549 cell suppressed the cell proliferation by increasing the amounts of G1 cells, and induced cell apoptosis. CONCLUSION: These results demonstrated the effectiveness of siRNA in lung caner cell, which indicated a possible clinical therapy method for lung cancer.
Key concepts: A549 cell, Transfection, Small interfering RNA, RNA interference, Molecular biology, Cell growth, Cell, Flow cytometry