ABSTRACT
<p><b>OBJECTIVE</b>To construct a short hairpin (sh)RNA targeting the gene encoding the MDM2 oncoprotein in order to investigate its role in human hepatocellular carcinoma (HCC) and its potential for use as a gene therapy strategy to inhibit HCC growth in vivo.</p><p><b>METHODS</b>Small interfering (si)RNAs were designed targeting the MDM2 gene (siMDM2-1 and siMDM2-2) and unrelated sequences (negative control) and cloned into the expression plasmid pGCSilencer-U6-neo-GFP. A HCC mouse model was established by subcutaneous inoculation of HepG2 cells (2 x 10(6) in 0.2 ml) into 20 nude mice. The inoculated mice were divided into four equal groups for tumor-localized injections of saline, negative control siRNA plasmid, siMDM2-1 plasmid, and siMDM2-2 plasmid. Tumor growth was observed daily (by caliper measurement) for one month, when mice were sacrificed by cervical dislocation. The tumor mass was resected for analysis of tumor inhibition rate (% = [(average tumor weight of control group - average tumor weight of treatment group) / average tumor weight of control group x 100]) and effects on MDM2 and p53 mRNA and protein expression (by reverse transcription- PCR and western blotting, both normalized to beta-actin). Significance of between-group differences was assessed by one-way ANOVA or LSD test; pairwise comparisons were made by the Chi-squared test.</p><p><b>RESULTS</b>siMDM2-1 and siMDM2-2 suppressed the xenografted tumor growth remarkably (60.6% and 54.6% inhibition rates, respectively), significantly reduced the expression ofMDM2 gene (62.8% and 61.6%) and protein (60.7% and 59.5%), and significantly increased p53 gene (47.1% and 45.6%) and protein (45.9% and 44.3%) (all, P < 0.05).</p><p><b>CONCLUSION</b>shRNA-mediated silencing of the MDM2 gene effectively inhibits HCC tumorigenesis of subcutaneously xenografted HepG2 cells in nude mice, and the mechanism may involve p53.</p>
Subject(s)
Animals , Humans , Male , Mice , Carcinoma, Hepatocellular , Genetics , Pathology , Cell Proliferation , Hep G2 Cells , Liver Neoplasms , Genetics , Pathology , Mice, Nude , Plasmids , Proto-Oncogene Proteins c-mdm2 , Genetics , Metabolism , RNA Interference , RNA, Messenger , Genetics , RNA, Small Interfering , Transfection , Tumor Suppressor Protein p53 , Metabolism , Xenograft Model Antitumor AssaysABSTRACT
Objective To use Excel worksheet function to achieve statistical analysis of the fourfold table qualitative data,and to improve knowledge of the.majority of researchers on the statistical capabilities of Excel.Methods The result of actual research published in Chinese Journal of Endemiology was the data source,and statistical analysis results of fourfold table data from Excel and from SAS software were contrasted.Results The worksheet function of Excel could realize the analysis fourfold table data chi-square test,obtained an accurate P values,but not just P ＞ 0.05 or P ＜ 0.05.The results of chi-square test of Excel were the same as the results from SAS software,but Excel could not obtain probability value of fisher exact probability.Conclusion As a dependent statistical analysis software,Excel is an easy to learn,easy to use,efficient and a good helper.In the absence of professional statistical software,Excel can be used to achieve data statistical analysis.