Detection Of P16mRNA, P16^INK4A And ZPPS Expressions In The Cacinogenesis Of Cervical Squamous Cell Carcinoma By Manual Tissue Chip

Objective: Cervical cancer is one of common malignant tumors which influence women's life and healthy. Worldwide, incidence of almost half a million new cases annually, 80% of them in developing countrys. It have become one of common malignant tumors and main reason of death. Death rate is second only breast cancer. Screening for cervical cancer widely available has reduced its incidence. Pathogenesis of cervical cancer is associated with many factors and phases. In the past decades, laboratory and clinical researches into different aspects of cervical carcinoma had been performed, but the carcinogenesis of cervical carcinoma is still not clear enough.p16 is a multiple tumor suppressor, it is the first anti-oncogene which direct effect cell cycle to suppression cell division. What kind of effection it is in the cervical cancer occurrence process has become the hot spot of the research. Many researchs have confirmed that the P16 protein expression unusual in the cervix carcinogenesis process, but its expression is elevates, or reduces, the view not one. Actually, What the deactivation mechanism of the p16 gene in the carcinoma of cervix occurrence process, p16 gene becomes one of the most important anti-oncogene in the cervical cancer studies. There are many questions between p16 and occurrence process of the cervical cancer.ZPP is one kind the glycoprotein which withdraws from the lower forms of plant. ZPP can stimulate the tumor cell multiplication. The research indicated that it is plant-related-person tumor antigen. Yueqi Zou passed through further to this antigen protein structure as well as the correlation gene research, continues to epurate it to made the ZPPS. The research had indicated that ZPPS after ¹³¹I mark obviously higher than the normal tissue in the mouse malignant tumor in radionuclide, also the expression speed and the degree are higher than ¹³¹I mark ZPP. But the research of the expression of ZPPS has not seen reported in the cervical cancer.This topic uses the tissue chip, In situ hybridization to research p16 mRNA, and uses the immunohistochemistry to research the expression ZPPS and P16 protein in normal cervical squamous epithelia, CIN and cervical carcinoma. To discusses the relations between p16 mRNA and biology behavior in normal cervical squamous epithelia, CIN and cervical carcinoma. To discuss the function of expression of p16 mRNA in the cervical cancer occurrencing and developing. To discuss the expression of p16 gene in the protein level and the mRNA level whether does tall. To discuss the p16 gene in the protein level and the molecular level in the development from the normal cervical squamous epithelia, CIN to cervical carcinoma.Methods:1 39 cases of normal cervical squamous epithelia, 154 cases of CIN and 54 cases of cervical carcinoma were collected.2 By manual tissue chip technique, this is done by using a needle to biopsy a standard histologic section and placing the core into an array on a recipient paraffin block, then fusing, slicing, gaining and baking.3 Immunohistochemistry SP method was used to examine for the expression, relation and the effect of p16 and ZPPS in cervical carcinoma, CIN and normal cervical squamous epithelia.4 In situ hybridization (ISH) was used to detect p16 mRNA in 200 cases of cervical samples, including 48 cases of cervical carcinoma, 119 cases of CIN and 33 cases of normal cervical squamous epithelia.5 Analysis the data use spss11.5, significance was set at p<0.05.Results:1 The result of tissue chip8 tissue chips (6×5,7×5) were made. Dots of tissue chip lined up in order, uniformed in size and no shifting. 8 tissue dots were absence, 39 tissue no significant tissue structures. The significant tissue structures were watched in the other dots. The rate of complete tissue dots was 83.3%⁹3.3%.The haematoxylin-eosin (HE) staining was uniformity and no dropping, moving and wrinkling.The immunohistochemical and ISH results show that the positive location was accurate and the background was clear.2 The expression of P16^INK4AIn normal cervical epithelia, the expression of P16^INK4A is absence. P16^INK4A expression is significant differentiated at normal cervical squamous epithelia with CINâ… , CINâ…¡, CINâ…¢and cervical invasive squamous cell carcinoma (P<0.01). P16^INK4A expression at CINâ… , CINâ…¡with cervical invasive squamous cell carcinoma is significant different (p<0.01), the expression at cervical invasive squamous cell carcinoma and CINâ…¢is no significant different(p>0.05), the expression at CINâ… , CINâ…¡and CINâ…¢is significant different (p<0.05).3 The expression of ZPPSIn normal cervical epithelia the expression of ZPPS is absence. ZPPS expression is no significant differentiated at normal cervical epithelia with CINâ… . There is significant differentiated at CINâ… , CINâ…¡with cervical invasive squamous cell carcinoma (p<0.05), but the expression of ZPPS is no significant differentiated at cervical invasive squamous cell carcinoma and CINâ…¢(p>0.05).4 The expression of p16 mRNAExpression of p16 mRNA in various lesions of cervix, 48 cases of cervical carcinoma, 119 cases of CIN and 33 cases of normal cervical seqamous epithelia were tested. The positive rates of p16 mRNA in normal cervical epithela, CINâ… , CINâ…¡, CINâ…¢and cervical carcinoma group were 18.18%, 43.75%, 56.41%, 75%, 79.17% respectively. p16 mRNA expression is significant differentiated at normal cervical epithelia with CINâ…¡, CINâ…¢and cervical invasive squamous cell carcinoma (p<0.01). There is significant differentiated at cervical invasive squamous cell carcinoma with CINâ… , CINâ…¡(p<0.01), but the expression at CINâ…¢and cervical invasive squamous cell carcinoma is no significant different (p>0.05).5 The relation of p16 mRNA,ZPPS,P16^INK4AThere was a positive correlation between expression of P16^INK4A and p16 mRNA(p<0.01), expression of p16 mRNA and ZPPS (p<0.05), expression of P16^INK4A and ZPPS.6 The relationship between the expression of p16 mRNA,P16^INK4A,ZPPS and clinical pathological features of cervical carcinoma.No significant relationship were found between the expression of P16^INK4A and clinical pathological features, but clinical stage. There was significant relationship between the expression of p16 mRNA and clinical stage. Significant relationship were found between the expression of ZPPS and metastasis.Conclusions:1 P16^INK4A overexpression rates become higher and higher from normal cervical squamous epithelia, CIN to cervical invasive squamous cell carcinoma.2 There was expression of p16 mRNA in normal cervical squamous epithelia. The positive rate of p16 mRNA from CINâ… , CINâ…¡, CINâ…¢to cervical invasive squamous cell carcinoma becomes gradually more higher.3 There was a positive correlation between overexpression of p16 and p16mRNA in occurrence and development cervical squamous cell carcinomas.4 ZPPS overexpression rates become higher and higher from normal cervical squamous epithelia, CIN to cervical invasive squamous cell carcinoma.5 There was a positive correlation between overexpression of P16^INK4A and ZPPS. Probably significant correlation of P16 and ZPPS in diagnose cervical squamous cell carcinomas.6 There was no significant relationship between overexpression of p16 and clinical pathological features, but clinical stage. There was significant relationship between expression of ZPPS and metastasis. Significant relationship were found between the expression of p16 mRNA and clinical stage.7 Manual tissue chip technique is simple, cost-effective, and reliable. This technique can provide a highly efficient, high-throughput mechanism for some research.