| Objective: Cervix cancer is the second most common cancer amongwomen worldwide. It is Serious harm to women’s health.According to WHOstatistics,it is much more common in developing countries, where83%ofcases occur and where cervical cancer accounts for15%of female cancers. Indeveloped countries, cervical cancer accounts for only3.6%of femalecancers.And in recent years, The morbidity and mortality of the Cervix cancerin young women were marked uptrend.Pathogenesis of cervical cancer isassociated with many factors and phases,but it is still unclear enough. So, thedomestic and foreign scholars pay attention to research the etiology andpathogenesis of cervical cancer.AKT protein is a kind of serine/threonine protein kinase. It is located inthe key part of phosphiti-dylinositoI3-kinases/protein kinase B signaltransduction pathway, which is the main effect molecules in downstream ofthe pathway. It is plays an important role in malignant tumor cell proliferation,angiogenesis, resistance of radiation and chemotherapy. P-AKT (phosphorylati-on AKT) is the activation form of AKT, which is able to activate thepathways downstream factor and play a central role in a variety of oncogenicprocesses including increase tumor cell proliferation, blockage of apoptosis,and activation of metastasis,invasion.BTAK (also known as Aurora A/Stk15/Aurora2) is a kind ofserine/threonine protein kinase. It occurs in multiple human tumour cell. Themain functions are responsible for centrosome maturation, spindle assemblyand mitotic enter and exit. It is considered as a critical factor in the centrosomematuration, spindle assembly. The amplification and overexpression of BTAKinduce centrosome duplication-distribution abnormalities and aneuploidy, anddaughter cell get abnormal number chromosome, which lead to heteroploid. Heteroploid cause tumor suppressor gene deletion, oncogene gain oractivation and eventual cause tumor formation. Some study indicate thatBTAK lead to malignant transformation by chromosome instability, activationAKT and signaling pathways downstream factor.In this study, we detect the the expression of AKT, P-AKT and BTAKprotein in normal cervical squamous epithelia, CIN and cervical squamous cellcarcinoma (CSCC) by the tissue chip and immunohistochemistry (IHC). Toanalysis mutual function and relations of AKT, P-AKT and BTAK abnormalexpression in cancer progression.To study interrelations and clinicalsignificance among AKT, P-AKT, BTAK and clinical indicators of cervicalcancer. It has great significance to comprehend the molecular biologymechanism of CSCC and improve the diagnosis rate. To discuss the clinicalsignificance will provide theoretical basis for clinical prevention, prognosisassessment and treatment of CSCC.Methods:531cases of normal cervical squamous epithelia,124cases of CIN(CINⅠ32cases, CINⅡ47cases, CIN Ⅲ45cases)and48cases of CSCCwere collected in Hebei general hospital.6Put200cases cervical tissues by manual tissue chip technique into arecipient paraffin block, then fusing, slicing, gaining, baking, and madeinto tissue chip.7To detect the expression of AKT, P-AKT and BTAK in normal cervicalsquamous epithelia, CIN and CSCC by IHC (Evsion method).8SPSS13.0was applied to analyze the results of experiment.Results:1The tissue chip preparation2Tissue chips (8×8) and1tissue chips (8×9) were made. Sectionthickness4-5μm, tissue dot matrix lined up in order and no shifting.6tissuedots were absence,9tissue no significant tissue structures. We get meaningfulsample185cases, including normal cervical squamous epithelia29cases,CINⅠ30cases, CINⅡ41cases, CINⅢ41cases and48cases of CSCC. The observation rate was90%.The haematoxylin-eosin (HE) staining wasuniformity, no dropping, moving and wrinkling. The IHC results show that thepositive location was distinct and the background was clear.2The expression of AKT in normal cervical squamous epithelia and variouscervical lesions.To detect the expression of AKT in44cases of cervical carcinoma,112cases of CIN and29cases of normal cervical squamous epithelia by IHC.There was no expression in normal cervical squamous epithelium. Theexpression of AKT was have obvious stratification, the positive cells werelocated in the1/3layer in low-grade CIN, and high-grade of CIN may be morethan the2/3or the full thickness. The positive rate from normal cervicalepithelia0%(0/29), CINⅠ20.0%(6/30), CINⅡ78.0%(32/41), CINⅢ80.4%(33/41) to CSCC81.8%(36/44). There were significant difference amongthose groups (p<0.05). AKT expression is significant difference among normalcervical epithelia and CINⅠ, CIN, CINⅢ, SCSS (p<0.05). AKT expression issignificant difference among CINⅠand CINⅡ, CINⅢ, CSCC (p<0.05).Thereis no significant difference between normal cervical epithelia and CINⅠ(P>0.05). AKT expression among CINⅡ, CINⅢ and SCC is no significantdifference too (P>0.05). The study of AKT and clinic parameters of CSCCsuggest that the expression of AKT were correlated to age, tumor size, clinicalstage, pathological rank and lymph node metastasis, but not to pathologicgrade.3The expression of P-AKT in normal cervical seqamous epithelia andvarious cervical lesions.The postive expression of P-AKT located in nuclear. The expression ofP-AKT from normal cervical epithelia65.％(19/29), CINⅠ73.3%(22/30),CINⅡ97.6%(40/41), CINⅢ89.4%(35/41) to SCC70.5％(31/44). Therewere significant differences among those groups (p<0.05). The expressions ofP-AKT was higher in CINⅡ, CINⅢ, CSCC than normal cervical seqamousepithelia and the difference was significant (P<0.05). There were significantdifferences between CINⅠand CINⅡ, CINⅡ and CSCC. The study of P-AKT and clinical pathological features of CSCC suggest that the expressionof P-AKT were correlated to pathologic grade and lymph node metastasis (P<0.05), but not to age, tumor size, and clinical stage (P>0.05). The positiverate of P-AKT protein in CSCC became higher when the tumor differentiationwas poor (P<0.05). Positive rate of P-AKT in CSCC became higher when thelymph metastasis was postive (P<0.05).4The expression of BTAK in normal cervical seqamous epithelia andvarious cervical lesions.BTAK positive expression mainly in cell plasma, a small number ofpresent in the upper squamous epithelium, in normal cervical squamousepithelium, pale yellow and base layer of cells no stain. With CIN levelsincrease, positive express location and intensity increases. Cancer are almostpositive expression. The positive rates of BTAK from normal cervicalepithelia, CINⅠ, CINⅡ, CINⅢ and CSCC group were10.3%,50.0%,75.6%,80.5%,81.8%. There were significant difference among those groups (p<0.05).The positive rates of BTAK were significantly lower in normal cervicalepithelium than in CINⅠ, CINⅡ, CINⅢ, CSCC. Expression of BTAK issignificant difference among CINⅠ and CINⅡ, CINⅢ, CSCC (p<0.05).There was no significant differrence among CINⅡ and CIN Ⅲ, CSCC(P>0.05). There was no significant difference among CIN and CSCC (P>0.05).Expression of BTAK showed no significant correlation among the tumor size,pathological rank and clinical stage (P>0.05). However, showed significantcorrelation between age and lymph node metastasis.5The relationship among AKT, P-AKT and BTAK in normal cervicalseqamous epithelia and various cervical lesions.The expression of AKT protein was positively correlated to theexpression of P-AKT protein (r=0.557, P<0.05).The expression of P-AKTprotein was positively correlated to the expression of BTAK protein(r=0.311,P <0.05).The expression of AKT protein was positively correlated tothe expression of BTAK protein (r=0.276, P <0.05). Conclusions:1Detect positive expression of AKT protein by IHC from normal cervicalsquamous epithelium, CIN into CSCC, which showed that positiveexpression rate increase gradually. It is demonstrated that AKT play animportant role in cervical cancer occurrence, development and malignanttransformation. The expression of AKT proteins were correlated to age,the size of tumor clinical stage, and lymph node metastasis (P <0.01).Theexpression of AKT is connected with proliferation and metastasis ofCSCC.2The positive rate of P-AKT is connected with pathologic grade andlymph node metastasis of CSCC by IHC. It is indicated that P-AKT isconnected with malignant degrees and metastasis of CSCC. Combinedwith the results AKT, suggest that we can co-test the expression ofAKT/P-AKT in CSCC to judge malignant degree, predict the clinicalstages and lymph node metastases in clinical practice. It is help forprognosis assessment.3To detect BTAK protein by IHC, the positive rate was gradually higherfrom normal cervical squamous epithelia, CIN to CSCC. The expressionof BTAK is connected with age and lymph node metastasis of CSCC.4The expression of AKT, P-AKT, BAKT showed a significantly positivecorrelation in each other.It is suggested that BTAK lead to malignanttransformation by activtion PI3K/AKT/P-AKT signaling pathway.5Mannual intensive tissue chips is a simple, efficient and stable technicalmethod. |
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