Study On Expression And Significance Of C/EBPα Gene In The Tissue And Cell Of Cervical Cancer

Objective: To study the C/EBPαgene protein expression level and mRNA expression level and clinical significance between Cervical Squamous Cell Carcinoma tissues and chronic cervicitis tissues of Uigur women. Analyzing C/EBPαgene protein expression level and clinical significance between Cervical Squamous Cell Carcinoma tissues and chronic cervicitis tissues of Han women. To explore C/EBPαgene effects on the HeLa cell survival by MTT methods and analyzing C/EBPαgene influenc HeLa apoptosis by Annexin-Ⅴ/PI double staining flow cytometry method.Methods: Collect Uigur cervical paraffin-embedded tissues 207 cases (Uigur cervical squamous cell carcinomas tissues 112 cases, chronic cervicitis tissues of Uigur cancer tissues 95 cases) from the people hospital of Kashi between Jan. 2008 and Apr. 2009. It detected the C/EBPαgene protein expression level in cervical squamous cell carcinomas tissues and chronic cervicitis tissues of Uigur women by immunohistochemistry(S-P Method). To detect C/EBPαgene mRNA expression level between 5 patients of cervical squamous cells carcinoma and their corresponding cervical normal tissues by real time quantified RT-PCR experiment. Constructed the eukaryotic expression vector of C/EBPαgene pcDNA3.1,this recombinant vector were transfected into Hela cells by using liposome transfection. Then analyzing the number of cell growth by MTT Method and the cell apoptosis were studied by AnnexinV/PI double stainning flow cytometry.Results: (1)The C/EBPαprotein expression in cervical squamous cell carcinomas tissues of Uigur women compared with chronic cervicitis tissues of Uigur women were obviously decreased, there was significant difference(P<0.05); C/EBPαprotein expression in cervical squamous cell carcinomas tissues of Han women were increased comparing chronic cervicitis tissues of Han women. (2) Through chi square test,there was significant difference(P<0.05), the expression of C/EBPαprotein were positive correlated with the hierarchical pathology of the cervical squamous cell carcinoma of Uigur women, Rank correlation coefficient of Spearman r=0.200. However, the Han people have no correlation. (3) Through chi square test,there was significant difference(P<0.05), the expression of C/EBPαprotein were negative correlated with the clinical stage of the cervical squamous cell carcinoma of Uigur women, Rank correlation coefficient of Spearman r=-0.269. whereas the Han people have no correlation. (4) Among cervical squamous cell carcinoma tissues of 5 cases, C/EBPαgene expression at mRNA level of the maxlmum is 4.79 and the minimum is 1.90,the average expressed level is 3.07±1.04. However, among their corresponding cervical normal tissues of 5 cases, C/EBPαgene expression at mRNA level of the maxlmum is 9.75 and the minimum is 2.46,the average expressed level is 5.63±2.98. Through test of homogeneity of varianc,mean square interclass is irregularity,undertake similar to t analysis,t=-3.15,p=0.006, it indicates that there was significant difference(P<0.05) between cervical squamous cell carcinomas tissues and corresponding cervical normal tissues of Uigur women. (5) It was successfully constructed the eukaryotic expression vector of C/EBPαgene pcDNA3.1,this recombinant C/EBPαpcDNA3.1 plasmid, pcDNA3.1 plasmid were transfected into Hela cells by using liposome. After C/EBPαgene of pcDNA3.1 constructs transfected into HeLa cells, the survial of HeLa cells were lower than pcDNA3.1 plasmid transfected in HeLa cells and non-transfected HeLa cells by MTT experiment, there were significant difference(P<0.05). After C/EBPαgene of pcDNA3.1 constructs transfected into HeLa cells, the apoptosis rate of HeLa cells were higher than pcDNA3.1 plasmid transfected in HeLa cells and non-transfected HeLa cells by Annexin-Ⅴ/PI double staining flow cytometry method,there were significant difference in three groups.Conclusion:1.The protein expression level of C/EBPαgene is significant decreased in cervical squamous cell carcinomas tissues of Uigur and Han women.2. The protein expression level of C/EBPαgene were positive correlated with the hierarchical pathology of cervical squamous cell carcinoma of Uigur women, However, the Han women have no correlation.3. The protein expression of C/EBPαgene were negative correlated with the clinical stage of cervical squamous cell carcinoma of Uigur women, However, the Han women have no correlation.4. The mRNA expression level of C/EBPαgene in cervical squamous cell carcinoma of Uigur women is obviously lower than their corresponding cervical normal tissues.5. Successfully constructed the eukaryotic expression vector of C/EBPαgene and transfected into HeLa cells,it indicated C/EBPαgene may inhibit HeLa cell growth and lead to HeLa cell apoptosis.