Objective To construct the RNA interference(RNAi) eukaryotic expression vectors(pSUPERpuro-RIP) targeting to the human RIP gene and check the interfering effects after transfecting into Huh7 cell line with these vectors.Methods Four pairs of 60 nt DNA strands containing specific sequences targeting to human RIP mRNA were designed and synthesized,then annealed and ligated with the linear eukaryotic [...]
15
May
15
May
Construction and Identification of Recombinant Adenovirus Vector of siRNA Targeting Skp2
Objective To construct an adenovirus vector expressing small interfering RNA(siRNA) targeting to Skp2.Methods The siRNA containing DNA sequence targeting to Skp2 and its negative control sequence were designed,synthesized,annealed and subcloned into pShuttle-H1 containing the green fluorescent protein.The linearized shuttle plasmid by Pme I was recombined with back-bone pAdEasy-1 in BJ5183 bacteria.The recombinant pAd-Skp2-siRNA adenovirus particles [...]
15
May
Influence of MACC1 Gene expression on the proliferation and migration of SBC-5 lung cancer cells
Objective:To investigate the influence of siRNA targeting metastasis-associated in colon cancer l(MACC1) gene on proliferation and migration of human lung cancer cell line SBC-5 in vitro.Methods:The siRNA targeting MACC1 gene and negative control siRNA were chemically synthesized,and were then transiently transfected into human lung cancer cell line SBC-5 by LipofectamineTM 2000.Expression of MACC1 mRNA and [...]
12
May
Effects of survivin siRNA on angiogenesis of human breast cancer xenografts in nude mice
Objective To investigate the effects of survivin siRNA on angiogenesis of human breast cancer MCF-7 cell xenografts in nude mice.Methods The animal model of tumor xenograft was established by transplantation with MCF-7 cells into nude mice.The models were randomly assigned to 3 groups: PcDNA3-Survivin siRNA treatment,PcDNA treatment and control group,which were respectively injected with PcDNA3-siRNA [...]
11
May
The effect of the small interfering RNAs for macrophage migration inhibitory factor on apoptosis of colorectal cancer cell line
Objective To investigate the effective mechanisms of the small interfering RNAs(siRNA) for macrophage migration inhibitory factor(MIF) on the apoptosis of mouse colorectal cancer cell line,CT-26.Methods CT-26 cells were treated with specific MIF siRNA as the experimental group,treated with nonspecific siRNA as the control group,and treated without siRNA as the blank group.Cell apoptosis was determined by [...]
11
May
Construction and identification TRF2 siRNA expression vector on the breast cancer MCF-7 cells
Objective:To construction adenovirus-mediated RNA interference vector expressing TRF2 small interfering RNA(rAd-shRNA-TRF2),and explore the specific inhibitory effect of this vector on TRF2 mRNA expression in MCF-7 cell.Methods:The recombinant adenovirus rAd-shRNA-TRF2 was obtained,and the vector was transfected into MCF-7 cells,then the expression of TRF2 mRNA and protein at 48 h after transfection was detected by FQ-RT-PCR and [...]
9
May
siRNA targeting Livin decreases tumor in a xenograft model for colon cancer
AIM: To evaluate the effect of silencing Livin gene expression with siRNA to apoptosis and proliferation in a colon cancer cell line.METHODS: To investigate the anticancer effect of silencing Livin gene expression, we established an siRNA transfected cell line using the HCT116 colon cancer cell line. After confirming the successful transfection, MTT assay, flow cytometry [...]
8
May
Current status and development of miRNA and siRNA research on gastric cancer
RNA interference(RNAi) is an important topic of epigenetics research in post-genome period.RNAi works as a post-DNA replication regulator for gene expression,and it is related to the occurrence and development of malignant tumors.The most usual participators of RNAi are MicroRNA(miRNA) and small interference RNA(siRNA).This review summarizes the basic theory of miRNA and siRNA,and provides recent progresses [...]
8
May
siRNA silencing RhoC expression induces apoptosis of human hepatocellular carcinoma BEL7402 cells
Objective:To study the effect of siRNA silencing RhoC expression on apoptosis in human hepatocellular carcinoma BEL7402 cells and the related mechanism,so as to provide an experimental evidence for liver cancer gene therapy.Methods: RhoC-siRNA eukaryotic vector pU6mRFP RhoC-siRNA was constructed and transfected into BEL7402 cells,and the transfection efficiency was examined by confocal microscope.RT-PCR and Western blotting [...]
6
May
The inhibitory effect of RNA interference on STAT3 expression in liver cancer cell line SMMC7721
AIM: To investigate the chemosensitivity small interfering RNA(siRNA) on liver cancer cell line SMMC7721.METHODS: The siRNA sequences design based on signal transducers and activators of transcription 3(STAT3) gene,siRNA were transfected into SMMC7721 cells through liposome lipofectamineTM 2000.The expression inhibition of STAT3 gene in SMMC7721 cells was measured by real-time relative quantitative PCR.The cells growth inhibition [...]