In less than 10 years since its inception, RNA interference (RNAi) has had extraordinary impact on biomedical science. RNAi has been demonstrated to influence numerous biological and disease pathways. Development an...In less than 10 years since its inception, RNA interference (RNAi) has had extraordinary impact on biomedical science. RNAi has been demonstrated to influence numerous biological and disease pathways. Development and adoption of RNAi technologies have been prolific ranging from basic loss-of-function tools, genome-wide screening libraries to pharmaceutical target validation and therapeutic development. However, understanding of the molecular mechanisms of RNAi is far from complete. The purpose of this brief review is to highlight key achievements in elucidating the bio- chemical mechanisms of the RNA-induced silencing complex and to outline major challenges for the field.展开更多
MicroRNAs (miRNAs) are small endogenous non-coding RNAs which can specifically silence gene expression, and thereby alter cell and organism phenotype. Deregulation of miRNA expression has been discovered in a variet...MicroRNAs (miRNAs) are small endogenous non-coding RNAs which can specifically silence gene expression, and thereby alter cell and organism phenotype. Deregulation of miRNA expression has been discovered in a variety of tumors and it is now clear that they contribute to cancer development and progression. Previous studies have indicated that miRNAs are involved in developmental timing, cell proliferation, apoptosis, morphogenesis [1], antiviral defense 121, and tumorigenesis [3]. In cancer pathways, altered expression of tumor suppressive or oncogenic miRNAs can disrupt regulatory mechanisms normal. Altered miRNAs expression patterns have been observed in a variety of diseased tissues. Cervical cancer is the most common malignant tumor in female reproductive tract. Recently more and more study showed a large number of miRNAs were down-regulated or up-regulated in cervical cancer. Recent data revealed that miRNA-218 (miR-218) played important roles in tumor initiation and development. This review focuses on analysis of miR-218 and will provide some insight into the progress of cervical cancer.展开更多
OBJECTIVE To explore the inhibition of ACHN cells via shRNAexpression vector mediated cyclinE1 gene silencing.METHODS The shRNA targeting at cyclinE1 gene was designedand synthesized. By ligation, the fragment was ins...OBJECTIVE To explore the inhibition of ACHN cells via shRNAexpression vector mediated cyclinE1 gene silencing.METHODS The shRNA targeting at cyclinE1 gene was designedand synthesized. By ligation, the fragment was inserted intopGenesil-1-U6 to construct the recombinant plasmid pGenesil-1-U6-cyclinE1. The identified recombinant plasmid was introducedinto ACHN cells with lipofectamine 2000. The inhibition ofcyclinE1 mRNA and protein expression were analyzed by RT-PCRand western-blotting. MTT method was used for observing cellproliferation and drawing growth curve. The cell cycle and ratiosof apoptotic cell were assessed by flow cytometric detection. Theability of invasion and speed of cell migration were detected bytranswell chamber invasive models and cell scratch method.RESULTS The inhibition of expression of cyclinE1 in ACHN cellsmediated by recombinant vector (0.0933 ± 0.05) was significantlylower than that in the group of transfected with empty vector(0.8827 ± 0.04) and the control group (0.9021 ± 0.03) (P < 0.05).Flow cytometry showed that recombinant cells were blocked inthe G_1 phase and the apoptotic ratio was increased significantly(11.15 ± 4.00)% (P < 0.05). The curves of cell growth indicated thatthe proliferation of cell transfected with recombinant plasmid wasinhibited significantly compared with that in control group (P <0.05). The results of transwell and cell scratch suggested that theabilities of invasion and migration of the cells transfected withrecombinant plasmid were decreased conspicuously (P < 0.05).CONCLUSION The expression of cyclinE1 could be inhibitedsuccessfully by RNA interference induced by shRNA expressionvector. This consequently inhibits the cell growth and inducesapoptosis. Our study provided a preliminary result in searching ofRNA interference (RNAi) therapy for renal cell carcinoma.展开更多
Virus-induced gene silencing is regarded as a powerful and efficient tool for the analysis of gene function in plants because it is simple, rapid and transformation-free. It has been used to perform both forward and r...Virus-induced gene silencing is regarded as a powerful and efficient tool for the analysis of gene function in plants because it is simple, rapid and transformation-free. It has been used to perform both forward and reverse genetics to identify plant functional genes. Many viruses have been developed into virus-induced gene silencing vectors and gene functions involved in development, biotic and abiotic stresses, metabolism, and cellular signaling have been reported. In this review, we discuss the development and application of virus-induced gene silencing in plant functional genomics.展开更多
文摘In less than 10 years since its inception, RNA interference (RNAi) has had extraordinary impact on biomedical science. RNAi has been demonstrated to influence numerous biological and disease pathways. Development and adoption of RNAi technologies have been prolific ranging from basic loss-of-function tools, genome-wide screening libraries to pharmaceutical target validation and therapeutic development. However, understanding of the molecular mechanisms of RNAi is far from complete. The purpose of this brief review is to highlight key achievements in elucidating the bio- chemical mechanisms of the RNA-induced silencing complex and to outline major challenges for the field.
文摘MicroRNAs (miRNAs) are small endogenous non-coding RNAs which can specifically silence gene expression, and thereby alter cell and organism phenotype. Deregulation of miRNA expression has been discovered in a variety of tumors and it is now clear that they contribute to cancer development and progression. Previous studies have indicated that miRNAs are involved in developmental timing, cell proliferation, apoptosis, morphogenesis [1], antiviral defense 121, and tumorigenesis [3]. In cancer pathways, altered expression of tumor suppressive or oncogenic miRNAs can disrupt regulatory mechanisms normal. Altered miRNAs expression patterns have been observed in a variety of diseased tissues. Cervical cancer is the most common malignant tumor in female reproductive tract. Recently more and more study showed a large number of miRNAs were down-regulated or up-regulated in cervical cancer. Recent data revealed that miRNA-218 (miR-218) played important roles in tumor initiation and development. This review focuses on analysis of miR-218 and will provide some insight into the progress of cervical cancer.
基金supported by a grant from Major State Basic Research Development Program,China(No.2002CB513107).
文摘OBJECTIVE To explore the inhibition of ACHN cells via shRNAexpression vector mediated cyclinE1 gene silencing.METHODS The shRNA targeting at cyclinE1 gene was designedand synthesized. By ligation, the fragment was inserted intopGenesil-1-U6 to construct the recombinant plasmid pGenesil-1-U6-cyclinE1. The identified recombinant plasmid was introducedinto ACHN cells with lipofectamine 2000. The inhibition ofcyclinE1 mRNA and protein expression were analyzed by RT-PCRand western-blotting. MTT method was used for observing cellproliferation and drawing growth curve. The cell cycle and ratiosof apoptotic cell were assessed by flow cytometric detection. Theability of invasion and speed of cell migration were detected bytranswell chamber invasive models and cell scratch method.RESULTS The inhibition of expression of cyclinE1 in ACHN cellsmediated by recombinant vector (0.0933 ± 0.05) was significantlylower than that in the group of transfected with empty vector(0.8827 ± 0.04) and the control group (0.9021 ± 0.03) (P < 0.05).Flow cytometry showed that recombinant cells were blocked inthe G_1 phase and the apoptotic ratio was increased significantly(11.15 ± 4.00)% (P < 0.05). The curves of cell growth indicated thatthe proliferation of cell transfected with recombinant plasmid wasinhibited significantly compared with that in control group (P <0.05). The results of transwell and cell scratch suggested that theabilities of invasion and migration of the cells transfected withrecombinant plasmid were decreased conspicuously (P < 0.05).CONCLUSION The expression of cyclinE1 could be inhibitedsuccessfully by RNA interference induced by shRNA expressionvector. This consequently inhibits the cell growth and inducesapoptosis. Our study provided a preliminary result in searching ofRNA interference (RNAi) therapy for renal cell carcinoma.
基金supported by the National Transgenic Research Projects of China (Grant No. 2009ZX08009-026B)
文摘Virus-induced gene silencing is regarded as a powerful and efficient tool for the analysis of gene function in plants because it is simple, rapid and transformation-free. It has been used to perform both forward and reverse genetics to identify plant functional genes. Many viruses have been developed into virus-induced gene silencing vectors and gene functions involved in development, biotic and abiotic stresses, metabolism, and cellular signaling have been reported. In this review, we discuss the development and application of virus-induced gene silencing in plant functional genomics.