Expression And Significance Of PIK3CA Gene In Tissues Of Cervical Disease And The Relationship With The Infection Of HPV

ObjectivesCervical cancer is the second common malignant tumor all around the world which is less than breast cancer.In different counties and areas it has significant deviation.In the women in developing country,the disease rate is in the first place of malignant tumor.The new case of a particular disease of cervical cancer every year in the twentieth century, the The new case of a particular disease of our country is 131.5 thousand,which is almost one third of the whole world.Since age fifty,we generally carry out Pap smear and to cure and general investigation of cervical cancer which made the sickness rate and death rate breakdown obviously.But recent year because increase of infection of HPV,disease rate of cervical cancer has upgraded obviously,and goes to more youthful.So searching the cause of a disease and clearing-cut the pathogenesy is very important for the prophylaxis and early diagnosis.The lipid second messenger which is mediated by PI3K/AKT signal convery passageway is very important.It is important for the proceeding of cell cycle,cell differentiation,survival of cell and the cell's infiltrating and transferRecent years the research of the chromosomal change of cervical cancer also proved that PIK3CA is a oncogene of cervical cancer.The increase frequency of the copies of chromosome 3q26.3 is common,and PIK3CA gene locate on this point.Harris take PIK3CA as poker through FISH,detecting that the copies increased obviously in cell line of cervical cancer C-4I,CaSKi and ME-180.This also prove that PIK3CA gene induces cervical cancer with the help of HPV. There is no recerch about PIK3CA gene in cervical cancer,also no recearch of the dependability with the infection of HPV.This experiment will detect the expressing level of PIK3CA in cervical cancer at the level of mRNA,than binding the infecting status of HPV to discuss the contribution of PIK3CA gene in the carcinogenesis,development, transfusion ofcervical cancer.And can provide theory basement for the precaution,early diagnosis,healing judging prognosis of cervical cancer.Materials and MethodsSample:110 paraffin samples of pathological changes of cervical tissues and normal cervical tissues without radiotherapy,immumotherapy or chemotherapy,were collected from China Medical University,May 1997 to May 2005.CC(cancer of the cervix)30 cases,CINⅢteam 30cases,CINⅡ/CINⅠteam 30 cases,and a normal cervical skin tissue 20 cases.Reagent:p110αhybridization in situ test and measurement kit,HPV general type primer,ddH20,10*buffer,dNTP,Taq-E,Diaminobenzidine(DAB)purchased from Fu Zhou Mai Xin Biology Ltd.Corp.Self confected EDTA acid antigen repaired buffered liquid and 3%H2_O2 liquid.Experimental Methods:The experiment was conducted by hybridization in situ according to instructions of the reagent cases.Take already knowen positive vulv-squamous carcinoma as positive control,and used phosphate buffered saline(PBS) instead of the first antibody as negative comparison samples.Every sample is stained with Hematoxylin-eosin.And HPV is detected at the level of DNA.Results Determination and Image Analysis:Result Determination:brown-yellow particles which distributed in the cytoplasm of vulvar epithelia of different pathological changes mean positive PIK3CA expression.PCR:positive assessment denominator is corresponding length fragment appears strap.Statistical Analysis:1.The positive rate of PIK3CAmRNA between teams and detection rate of HPV DNA between teas use multiplicity chi square test.2.The relationship between positive rate of PIK3CAmRNA and detection rate of HPV DNA adopts consistency checking calculating KI,0.75＞KI＞0.4 indicating that the concordance is fine,KI＞0.75 indicating that the concordance is very good.3.The relationship between expression of PIK3CAmRNA,the infection of HPV and the histodifferentiation degree,clinical stage and absorbent gland adopts exact propability chi square test.Results一,The expression status of PIK3CA mRNA in tissues of cervix of diseaseThe positive rate of PIK3CA mRNA in cervical cancer,CINⅢ,CINⅡ/CINⅠand normal epithelium tissue is 66.7%,60%,16.7%,0.00%.the expression levels of PIK3CA in CCX and CINⅢwere significantly higher than those in normal cervical tissues and CINⅠ,CINⅡ(P＜0.01).There was no statistical significance between normal cervical tissues and CINⅠ,CINⅡas well as between CC and CINⅢPIK3CA at the expression(P＞0.05).In the CC group,the expression level of PIK3CA was associated with grade, clinical staging,but no relationship with lymph node metastasis status(P＜0.05).二,The expression status of HPV DNA in tissues of cervix of diseaseThe expression of HPV in group of CC,CINⅢ,CINⅡ/CINⅠand normal epithelium tissue is 83.3%,66.7%,56.7和0%,there is gradation between cervical cancer and CINⅡ/CINⅠ,normal cervical tissues(P＜0.01);and between CINⅢand normal cervical tissues(P＜0.01);and also between CINⅠand normal cervical tissues(P＜0.01), there are no difference between other teams(P＞0.05).The differences between teams of stageⅠ,ⅡandⅢ/Ⅳ,teams of well-differentiated and poorly differentiated and teams of having and not having lymph node metastasis has no significance.Conclusion1.Over expression of PIK3CA and the infection of HPV are associated with development and progression of the CACX. 2.In the happening and developing of cervical cancer the infection of HPV is associated with over expression of PIK3CA..3.PIK3CAmRNA can be a monitoring index of the happening and developing of cervical cancer and the infection of HPV can easily lead to cervical cancer.