The Expression Of Runx3 In Cervical Cancer And Its Proliferation Effect On Hela

Background:Cervical cancer is one of the most common malignancies of women in the developing countries, and the incidence rate of it has become the first female reproductive system cancer, cervical cancer has become a serious threat to women's health. In the recent years, With the changes in people's social life style in recent years, the patients who suffered from cervical cancer become younger and younger significantly, which can not be ignored. It is a long-term, complex developed process from normal cervical epithelium to cervical intraepithelial neoplasia and cervical cancer, the study of its etiology has been hot at home and abroad, and its pathogenesis has not been fully understood yet. The Scholars have agreed that there is a strong connection between human papillomavirus infection and the development of cervical cancer. The etiopathogenisis of the cancer is quite complicated which is generally considered to be of multiple factors. Many discoveries have shown that cervical cancer incidence and tumor suppressor genes are closely related.Human runt-related transcription factor 3 (RUNX3) is a new tumor suppressor gene discovered recently, it is a member of the RUNX transcription factor family. Many studies have found that RUNX3 gene expression lever was significantly lower in tumor tissues than in normal tissues, like in gastric cancer, lung cancer, pancreatic cancer, hepatocellular carcinoma and colorectal cancer etc, and it correlated with the degree of tumor differentiation. In the more malignant tissue, the RUNX3 gene expression lever is lower. The research of RUNX3 gene in tumor occurrence, development, metastasis and prognosis has become very hot at home and abroad. RUNX3 gene silenced or reduced is associated with the incidence of cancer genesis, and its expression level is closely related to cell differentiation, cancer metastasis. The relationship between RUNX3 expression and cervical cancer has not reported yet. This research has shown that RUNX3 gene promoter CpG island hypermethylation is the main deactivation mechanism. 5-Aza-2'-deoxycytidine (5-Aza-CdR) is a DNA methyltransferase inhibitor, it can make the tumor suppressor gene re-expression and restore tumor suppressor function through demethylation.Therefore, this subject focused on RUNX3 tumor suppressor gene expression in cervical cancer, and reveals its role in the development of cervical carcinoma. We use the5 - aza -2 '- deoxycytidine which can restore the expression of RUNX3 gene to further explore the function of RUNX3 gene in the Hela cells.Objective:1. To identify RUNX3 gene expression level under different pathological differences, and to explore its relationship with cervical cancer;2. By 5-aza -2 '-deoxycytidine which is a DNA methyltransferase inhibitor induced RUNX3 gene expression in Hela cells, we can find the changes of proliferation and apoptosis in Hela cells, investigate the relationship between RUNX3 gene and the occurrence of cervical cancer, and provide experimental evidence for the early diagnosis and future treatment.Methods:1. To detection the expression of RUNX3 gene in cervical cancer, cervical intraepithelial neoplasia (CIN) and normal cervical tissue by immunohistochemistry, western blotting and semi-quantitative RT-PCR.2. After the treatment of 5-aza -2 '-deoxycytidine we took semi-quantitative RT-PCR to detected the expression of RUNX3, and analysis biology changes by MTT, flow cytometry and colony formation of tumor cell,Results:1. Immunohistochemisty results suggested that RUNX3 showed lower expression in cervical cancer than in CIN and normal cervical epithelial,; the expression is declining in stageⅠ,ⅡandⅢ, also; the expression of RUNX3 protein is lower in lymph node metastasis tissues than without lymph node metastasis tissues, and all the difference is statistically significant (P<0.05);2. It confirmed that expression of RUNX3 was lower in cervical cancer compared with normal cervical epithelium by western blotting,and the difference is statistical significant (P<0.05);3. The expression of RUNX3mRNA was lower in cervical cancer compared with normal cervical epithelium by semi-quantitative RT-PCR experiment, and the difference is statistical significant (P<0.05);4. The Hela cells in the control groups failed to detect the expression of RUNX3 mRNA, but it was resumed after 5-Aza-CdR treatment and the expression was relative with the dose effect;5. The growth of Hela cells were significantly inhibited after the treatment of 5-Aza-CdR, with the increasing concentrations of the drug, the effect was more obvious;6. The apoptosis rate of the Hela cells compared with the control group was increased after the treatment of 5-Aza-CdR, and the difference is statistical significant (P<0.05);7. The ability of colony formation of Hela compared with the control group was reduced after the treatment of 5-Aza-CdR, and the difference is statistical significant (P<0.05).Conclusion:1. The expression of RUNX3 was reduced in the cervical tissues, and it related with the stage of tumor and the cell metastasis.2. The level of RUNX3 mRNA was activated after the treatment of 5-Aza-CdR in Hela cells, there caused a declining of cell proliferation and promoted cell apoptosis.