ABSTRACT
Objective To detect steroidogenic acute regulatory protein (StAR) expression in apolipoprotein E-deficient mice at different ages and serum lipid levels. Methods Nighty-six C57BL/6J and apoE-/- mice were enrolled, which were divided into 16 groups with 6 mice per group according to age (1 day, 1, 3, 5 months), sex and genotype (C57BL/6J and apoE-/-). The serum lipid levels in C57BL/6J and apoE-/- mice were detected by commercial kits. StAR mRNA and protein expressions in liver were detected by Real-time PCR and Western blot respectively. Results ApoE-/- mice had higher LDL-cholesterol and lower HDL-cholesterol compared with C57BL/6J mice of the same age and sex. StAR mRNA and protein expressions were decreased following with aging in C57BL/6J mice. However, in apoE-/- mice with higher lipid levels, StAR mRNA and protein expressions were changed with the lipid levels other than ages. StAR mRNA and protein increased in the early stage, and then decreased with the increasement of lipids levels. Conclusions StAR could affect lipids levels and may be an effective regulator for atherosclerosis and other cardiovascular diseases.
ABSTRACT
Objective RNA interference refers to post-transcriptional gene silencing caused by double strands RNA.To investigate the effect of EGFR receptor on esophageal carcinoma,the expression vector of HER4 gene targeted small interfering RNA was constructed to observe its silencing effect in human esophageal carcinoma cell line Eca-109,in order to find a promising method for the gene therapy of this disease.Methods Two complementary oligo DNA strands targeting HER4 gene were designed and synthesized according to the principles of designing siRNA.After annealing,oligo DNAs were inserted into SUPER.neo+gfp vector,then enzyme digestion analysis and DNA sequencing were applied.After transfecting it into human esophageal carcinoma cell line,we detected the level of expression of HER4 gene through real-time quantitative PCR and Western Blot.Results The enzyme digestion analysis and DNA sequencing show that HER4 gene targeted small interfering RNA and its expression vector were constructed successfully,and after transfection,the expression of HER4 gene in esophageal carcinoma cell line was suppressed greatly.Conclusion HER4 gene targeted small interfering RNA and its expression vector were constructed successfully,and could decrease the expression of HER4 gene in Eca-109 cell line,which laid the foundation for the following experiment.