The role of suppression of small RNA molecules in the management of malignity and viral infection on human was studies and discussed. SiRNA (small interference RNA) suppressing gene expression was described. In the year 2001, Ribopharma AG researchers had first demontrated the function of RNAi in mammal cells. SIRPLEX is appropriate with target gene, for using in the treatment of suppression of pathological gene in various genera, including human.
RNA ; Genes ; Gene Silencing

RNA interference (RNAi) is a post-transcriptional gene silencing process by targeting mRNA for degradation in a sequence-specific manner. This powerful platform has enormous potential in functional genomics and medical research. As a tool to knock out expression of specific genes in a variety of organisms, RNAi was used to investigate gene function in a high throughput fashion. Highly conserved in evolution RNAi appears to have evolved as a cellular defense mechanism in plants and animals to suppress viral infection, transposon jumping and endogenous aberrant genes. Exploiting the natural mechanism, the researchers can shut down disease-causing genes and develop novel therapeutics against infection, tumor and other disease.
Gene Expression Regulation ; Gene Silencing ; Genomics ; RNA Interference ; RNA, Small Interfering ; RNA-Induced Silencing Complex

The purpose of this review is to confirm the reason resulted the gene silence and explore the countermeasure avoiding the gene silence in transgene plant. The method is to divide the gene silencing into transcriptional gene silencing(TGS) and posttranscriptional gene silencing(PTGS). Several models resulted PTGS were analyzed by RNA threshold model, ectopic pairing and aberrant RNA model and ds-RNA model. The results showed that it was important to decide the phenomena of restraining transgene silencing and the mechanism of PTGS. The strategies of identification of gene function and prevention of virus were presented by RNAi and gene silencing respectively, etc.
Animals ; Gene Silencing ; Models, Genetic ; RNA ; RNA Processing, Post-Transcriptional

OBJECTIVETo find some effective short interfering RNA's sites targeting HBV surface gene sequence using shRNA expression vectors.

METHODSFour shRNA expression vectors targeting HBV surface gene sequence were constructed based on pAVU6 + 27 vector, and cotransfected into AD293 cells with HBs-EGFP fusion gene plasmid. The changes of HBs-EGFP image were detected by FACS and microscopy. The HBs-EGFP mRNA expression was evaluated by RT-PCR.

RESULTSFour shRNA expression vectors and HBs-EGFP fusion gene plasmid were successfully constructed. pAVU6 + 4sh579 vector inhibited the HBs-EGFP expression by 69.8% in AD293 and suppressed the HBs-EGFP mRNA expression by 74.6%.

CONCLUSIONSThe results showed that the 579 site of HBV surface gene sequence was an effective target and pAVU6 + 4sh579 vector could suppress the HBs-EGFP expression in AD293 cells

Gene Expression Regulation, Viral ; Gene Silencing ; Gene Targeting ; methods ; Hepatitis B Surface Antigens ; genetics ; metabolism ; Humans ; RNA Interference ; RNA, Small Interfering ; genetics ; RNA-Induced Silencing Complex ; genetics

OBJECTIVETo explore the biological function and possible underlying mechanism of aldo-keto reductase family 1 member B10 (AKR1B10) gene during hepatocarcinogenesis.

METHODSA pair of chemically synthesized small interfering RNA (siRNA) targeting on AKR1B10 was transfected into liver cancer cell line MHCC97H by LipofectamineTM 2000. After confirming the interfering effects of AKR1B10-siRNAs through Quant SYBR Green polymerase chain reaction (Real-time PCR), Western blot and enzymatic activity assay, the capabilities of proliferation and apoptosis of the transfected cells were observed by CCK-8 assay and flow cytometry analysis, and the expressions of a group of tumor-related gene such as c-myc, c-fos, N-ras were observed through Real-time PCR.

RESULTSThe expressions of AKR1B10 and the enzymatic activity were down-regulated significantly in AKR1B10-siRNA-transfected cells. Compared with mock and blank control groups, cell growth in AKR1B10-siRNA-transfected group was inhibited by 26.6%+/-3.1% at 72h after transfection. The ratio of apoptotic cells was 37.3%+/-1.0% in AKR1B10-siRNA-transfected group, which was significantly higher than that in mock and blank control groups (P < 0.01). Real-time PCR showed that the expressions of oncogene c-myc, c-fos and N-ras, and the proliferation-associated gene ki-67 were down-regulated in AKR1B10-siRNA-transfected cells, while the expressions of apoptosis-promoting gene caspas-3 and bax were up-regulated.

CONCLUSIONSAKR1B10 might promote proliferation, inhibit apoptosis and then induce malignant transformation of hepatocytes by regulating the expression level of some tumor-related genes.

Aldehyde Reductase ; genetics ; Cell Line, Tumor ; Gene Expression ; Gene Silencing ; Humans ; RNA, Small Interfering ; genetics

Color is an important indicator for evaluating the ornamental traits of horticultural plants, and plant pigments is a key factor affecting the color phenotype of plants. Plant pigments and their metabolites play important roles in color formation of ornamental organs, regulation of plant growth and development, and response to adversity stress. It has therefore became a hot topic in the field of plant research. Virus-induced gene silencing (VIGS) is a vital genomics tool that specifically reduces host endogenous gene expression utilizing plant homology-dependent defense mechanisms. In addition, VIGS enables characterization of gene function by rapidly inducing the gene-silencing phenotypes in plants. It provides an efficient and feasible alternative for verifying gene function in plant species lacking genetic transformation systems. This paper reviews the current status of the application of VIGS technology in the biosynthesis, degradation and regulatory mechanisms of plant pigments. Moreover, this review discusses the potential and future prospects of VIGS technology in exploring the regulatory mechanisms of plant pigments, with the aim to further our understandings of the metabolic processes and regulatory mechanisms of different plant pigments as well as improving plant color traits.
Plant Viruses/genetics* ; Plants/genetics* ; Gene Silencing ; Plant Development ; Gene Expression Regulation, Plant ; Genetic Vectors

Animals ; Gene Silencing ; physiology ; Gene Targeting ; Genetic Therapy ; methods ; Genomics ; Humans ; RNA Interference ; RNA, Small Interfering ; genetics ; therapeutic use ; RNA-Induced Silencing Complex

The stiffness of cancer cells is attributable to intermediate filaments such as keratin. Perinuclear reorganization via phosphorylation of specific serine residue in keratin is implicated in the deformability of metastatic cancer cells including the human pancreatic carcinoma cell line (PANC-1). 12-O-Tetradecanoylphorbol-13-acetate (TPA) is a potent tumor promoter and protein kinase C (PKC) activator. However, its effects on phosphorylation and reorganization of keratin 8 (K8) are not well known. Therefore, we examined the underlying mechanism and effect of TPA on K8 phosphorylation and reorganization. TPA induced phosphorylation and reorganization of K8 and transglutaminase-2 (Tgase-2) expression in a time- and dose-dependent manner in PANC-1 cells. These effects peaked after 45 min and 100 nM of TPA treatment. We next investigated, using cystamine (CTM), Tgase inhibitor, and Tgase-2 gene silencing, Tgase-2's possible involvement in TPA-induced K8 phosphorylation and reorganization. We found that Tgase-2 gene silencing inhibited K8 phosphorylation and reorganization in PANC-1 cells. Tgase-2 gene silencing, we additionally discovered, suppressed TPA-induced migration of PANC-1 cells and Tgase-2 overexpression induced migration of PANC-1 cells. Overall, these results suggested that TPA induced K8 phosphorylation and reorganization via Tgase-2 expression in PANC-1 cells.
Cell Line ; Cystamine ; Gene Silencing ; Humans ; Intermediate Filaments ; Keratin-8* ; Phosphorylation* ; Protein Kinase C ; Serine

This study examined the possible role of p120ctn in the pathogenesis and development of pancreatic cancer. PANC-1 cells, a kind of human pancreatic carcinoma cell line, were cultured in this study. p120ctn was immunocytochemically detected in PANC-1 cells. The recombinant lentivirus vector was constructed to knock down the p120ctn expression of PANC-1 cells. Real-time quantitative PCR (RQ-PCR) and Western blotting were used to determine the expression of p120ctn and E-cadherin in PANC-1 cells after p120ctn knockdown. The adhesion, invasion and migration capacity of PANC-1 cells after p120ctn knockdown was detected by cell adhesion, invasion and migration assays. Cell growth was measured by the MTT method. Cell cycle and apoptosis were analyzed by fluorescence-activated cell sorting. The results showed that p120ctn knockdown led to significantly down-regulated E-cadherin and a reduced cell-to-cell adhesion ability in PANC-1 cells. shRNA-mediated knockdown of p120ctn reduced invasion and migration capacity of PANC-1 cells, inhibited cell growth, caused a significant decrease in the percentage of cells in G(1), an increase in S, and promoted apoptosis of PANC-1 cells. It was concluded that p120ctn plays a pivotal role in the proliferation and metastasis of pancreatic carcinoma, suggesting that p120ctn is a novel target for pancreatic carcinoma treatment.
Catenins ; genetics ; Cell Line, Tumor ; Gene Silencing ; Humans ; Pancreatic Neoplasms ; genetics

Epigenetic modification, which involve DNA methylation, RNA-associated silencing and histone modification, is implicated in cell proliferation, differentiation, survival, apoptosis and malignant transformation. Some leukemogenesis has been shown to be aberrance of epigenetic modification. This paper discussed the potential causes of some of leukemias correlating with the methylation of cell cycle regulation genes, small interference RNA and modification abnormality of histone after translation. The study on epigenetic modification abnormality of leukemia cells provides a new strategy for treatment of leukemia.
DNA Methylation ; Epigenesis, Genetic ; Gene Silencing ; Histones ; Humans ; Leukemia ; genetics ; RNA, Small Interfering ; genetics