Using Tissue Microarry Method To Detect The Expression And Clinic Significance Of Survivin MRNA, Its Protein And P63 Protein In Cervix Carcinoma

Purpose: Cervical carcinoma is one of common malignant tumor in female genital tract,it experience a long time of lesion precancerous,which is cervical intraepithelial neoplasia. We knowe that it is related to abnormal generation and differentiation of cell,and also to the variation of apoptosis.The disbalance of generation and apoptosis can induce to tumor.This experiment wants to detect the protein of survivin and p63 in normal cervix,CIN and cervix squamous carcinoma,in order to find the role of survivin and p63 in the cervix cancer;the relationship between survivin and p63 with infiltration and metastasis;observing the dictinction of p63 in different cervix damage,in order to explain its role in the differentiation of cervix epithelium tissue;to reveal pathogenes- y of cervix cancer through the dependablity of survivin and p63.Method:1 The expression of survivin protein,p63 protein in 54 cases of cervix cancer,39 cases of normal cervix tiusse and 154 cases of CIN was detected by immunohistochemistry(IHC). 2 The expression of survivin mRNA in 54 cases of cervix cancer,39 cases of normal cervix tiusse and 154 cases of CIN was detected by in Situ Hybridization(ISH).3 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,slic- ing,gaining and baking.Result:1 The result of tissue chip6 tissue chips (6×5),and 2 tissue chips(7×5)were made. Dots of tissue chip lined up in order,uniformed in size and no shifting.51 tissue dots were no significance.So the actually cervix cancer 48 cases,normal cervix tissues 35cases,CIN 113 cases. The rate of complete tissue dots was 89%. The haematoxylin-eosin (HE) staining was uniformity and no dropping,moving and wrinkling.The immunohistochemical results show that the positive location was accurate and the background was clear.2 The expression of Survivin in different cervix lesionsThe expression of Survivin mRNA and protein in 48 cases of cervix cancer,35 cases of normal cervix tiusse and 113 cases of CIN were detected by ISH and IHC respectively.The results showed that in both mRNA and protein level,the expression of Survivin in cervix cancer (ISH:64.58%,IHC:87.50%) was significantly higher then in normal cervix tiusse(ISH:14.29%, IHC:31.43%)(P<0.05),the expression of Survivin protein betwwen cervix cancer and CINⅠ,Ⅱ,Ⅲwere no obviously different (P>0.05);the expression of Survivin mRNA in cervix carcinoma was significantly higher then in CINⅠ(29.63%,P<0.05),but have no obviously different between carcinoma and CINⅡ,Ⅲ(P>0.05).Results of ISH and IHC showed that the expression of Survivin wasn't correlated with lymph node metastasis,differe- ntiation degree and clinical stage(P>0.05).There was a close correlation between the expression of Survivin protein and Survivn mRNA in cervix carcinoma tissues(rs=0.529,P<0.05).3 Expression of P63 in different lesions of cervixThe expression of P63 protein in 48 cases of cervix cancer,35 cases of normal cervix tiusse and 113 cases of CIN were detected by IHC.The result showed that,the strong expression rate of P63 in cervix carcinoma(25%)was significantly higher than that in normal cervix tiusses,CINⅠ,Ⅱ,their strong expression rate respectively were 0%,3.70%,8.57%(P<0.05);but it wasn't relationship with CINⅢ(17.65%,P>0.05);CINⅠ→Ⅱ→Ⅲ,the expression of P63 had significant difference.The expression of P63 wasn't correlated with age,diamet- er of tumor,clinical stage and lymph node metastasis(P>0.05), but it was correlated with tumor differentiation(P<0.05).4 The relationship with P63 protein and Survivin proteinThere was a close positive correlation between the expression of P63 and Survivin protein in cervix carcinoma(rs=0.364,P<0.05).5 The relationship between P63 and Survivin mRNA There was no close correlation between the P63 protein and Survivin mRNA in the cervix carcinoma tissues(rs=-0.016,P>0.05).Conclusions:1 The expression of Survivin mRNA and protein in cervix carcinoma was significantly higher than that in normal cervix tissues,which indicated that Survivin could be correlated with uterine cervix cancer.The expression of Survivin mRNA in cervix cancer was significant different with that in CIN,so it was useful to morning lesion of cervix.The expression of Survivin mRNA and protein were no significant correlation with age,tumor differentiation,lymph node metastasis,tumor diameter and clinical stage.2 The Survivin protein had signification difference between normal cervix and cancer tissue,it illustrated that Survivin can use to detect cervical intraepithelial neoplasia;the expression of Survivin protein in cervix carcinoma had no relationship with cervical intraepithelial neoplasia,so Survivin can't participate the process of different cervix lesion.Survivin had obviously variation of layer,its disposition rule concorded with CIN,it cued that Survivin can provide a visualized basement to CIN.3 There was a close correlation between the expression of Survivin protein and Survivin mRNA in the cervix cancer tissues,it indicated that the expression of Survivin protein indirectly reflected the expression of Survivin mRNA,and the high expression of Survivin protein could be the result of high transcription of Survivin gene.4 The expression of P63 in cervix cancer was significantly h- igher than that in normal cervix tissues,which indicated that P63 could be related with cervix cancer.In CINⅠ→Ⅱ→Ⅲand different differentiated tumor tissues,the expression of P63 had significance different,it illustrated that P63 had inverse ratio with differentiation degree of squamous epithelium,also cued that it was uesful to the stages of CIN;the expression of P63 were norelationship with age,lymph node metastasis,tumor diameter and clinical stage.5 There was a close relationship between the expression of Survivin protein and P63 protein in cervix carcinoma tissues,along with the expression of Survivin strengthening,the expression of P63 protein also enhanceing,it indicated that there could be a positive cooperation between Survivin protein and P63 protein.But the expression of P63 protein had correlation with Survivin mRNA,it indicated that P63 and Survivin coordinated on the protein level.6 Mannual tissue chip technique is simple,cost-effective,and reliable.This technique can provide a highly efficient, high-throughput mechanism for some research.