多糖抑制胃癌細(xì)胞機(jī)制及熒光PCR分析apoE基因型.pdf

1、中南民族大學(xué)碩士學(xué)位論文多糖抑制胃癌細(xì)胞機(jī)制及熒光PCR分析apoE基因型姓名：嚴(yán)冰冰申請(qǐng)學(xué)位級(jí)別：碩士專業(yè)：生物化學(xué)與分子生物學(xué)指導(dǎo)教師：覃瑞;游上游20080501多糖抑制胃癌細(xì)胞機(jī)制及熒光 PCR 分析 apoE 基因型 II Abstract 1.In this study, transformations of the cultured human gastric cancer cells (SGC7901) treatin

2、g with polysaccharide were investigated. The results showed that multiplication of the cultured cells was restrained by treating with polysaccharide, and more cells multiplication was restrained because of increasing i

3、n the polysaccharide dosage and treating time. The cultured cells performed typical apoptosis both in morphological characters and DNA fragmentation after treating 48 hours with 10mg/mL polysaccharide. There were abou

4、t 40% of the cultured cells performed apoptosis, and expression of BCL-2 protein was down-regulated and BAX protein was up-regulated. So, treating with polysaccharide can induce apoptosis and restrain cell multiplicati

5、on, which is used for human gastric cancer therapy. 2.To establish a new effective single nucleotide polymorphism (SNP) typing method by using real-time polymerase chain reaction (PCR) for identifying apolipoprotein E

6、 genotypes. To determine Cys112Arg locus alleles of human apolipoprotein E genetic polymorphisms, genotypes were assigned based on PCR melting curve analysis with the indicating fluorophore SYBR Green I. In order to deve

7、lop more reliable SNP assays, we use high fidelity Taq polymerase. The accuracy of SNP typing determined by real-time PCR was validated by comparison with results from direct DNA sequencing. Each sample was determined by

8、 double measuring tubes, and two melting curves were analysis. Compared the Tm value of the sample with the Tm of the standard substance, we conclude the apoE genotype of the samples. The results of 30 samples for apoE g

9、enotype are E3/3(27/30)and E3/4（3/30）, which is accordant with the results from PCR-RFLP and DNA sequencing. The presented allelic assay was specific, easy to operate and applicable for apolipoprotein E genotyping of hu