Expressions Of Omi/HrA2in Oral Squamous Cell Csrcinomas And Their Significance

Objective:Oral squamous cell carcinoma is the most common malignanttumor in the oral mucosa, about80%of the oral mucosa malignant tumorpathological type for oral squamous cell carcinoma, accounts for about40%ofthe head and neck cancer, but its pathogenesis is not clear, diagnosis andtreatment means are single so further understand oral squamous cellcarcinoma of the pathogenesis and on the basis to explore early diagnosis andearly treatment is very urgent.HtrA2((High temperature requirement A, HtrA)is one of serine-proteinasas which exist in body of bacteria.In2000,Researchers isolated a serine protease in mammals and named it Omi. Inthe same year,Savopoulos refined the enzyme by baculovirus expressionsystem and named it HtrA2.Some researchers have shown that Omi/HtrA2ishighly expressed in many human cancer cell lines, such as liver cancer,gastriccancer,prostatic carcinoma,laryngocarcinoma and so on.While, therelationship between Omi/HtrA2in the human cancer cell high expression andthe cancer cell apoptosis,proliferation and growth are not fully understoodyet. The aim of our research is to test the expresstions of Omi/HtrA2in oralsquamous cell carcinoma and their significance to gain the relationshipbetween Omi/HtrA2and the Oral squamous cell carcinoma occurrence,andoral squamous cell carcinoma patients with early diagnosis and early treatment,malignant degree analysis and prognostic.Methods：Collect50cases of oral carcinoma specimens, paracanceroustissue,epithelial dysplasia and normal tissues cut off by Surgical resectionduring October2010to June2011, All patients did not take any therapy, thespecimens are divided into two groups immediately after they are cue off, oneis immobilized in4%formaldehyde, then Embed it by Paraffin, the other isstored in-80℃. 1The expressions of Omi/HtrA2are examined by Immunohistochemistry in50cases of oral carcinoma specimens, paracancerous tissues,epithelialdysplasias and normal tissuesTake wax blocks of each group, dewaxing to water, operate accord toMaxvision2/HRP kit instructions, masculine of Omi/HtrA2stanining can beobersered Brown granules in cytoplasm through microscopic,analysis if thereis a statistically significant among the different groups.2The expressions of Omi/HtrA2are examined by RT-PCR in50cases of oralcarcinoma specimens, paracancerous tissues,epithelial dysplasias and normaltissues.Extract the total RNA through the method of Trizol. The total RNAextracted was identified by agarose gel electrophoresis and was definedquantitatively by UV spectrophotometer. Compound the extracted RNA intothe first chain of cDNA according to the instructions of reverse transcriptase.Amplificate the purpose gene using the gapdh as the baction.Use gel imagingsystem for observation and image acquisition. Electrophoresis scanning theresults, in their place is visible on the specific DNA bands and analysis theresults by Quantity One software.3Results criteria3.1ImmunohistochemistryMasculine of Omi/HrA2staining can finding Brown granules incytoplasm, its expression is evaluated by Immunohistochemicalsemi-quantitative method.3.2Semi-quantitative analysis of gene expressionScanning the gray of GAPHD and objective products in order to gain therelative expression.4Statistical MethodsStatistical analysis were performed using statistical package SPSS13.0,χ~2test、SNKand analysis of variance were used. P<0.05was accepted asstatistically significant. Results：1The expression of Omi/HtrA2in oral carcinoma specimens, paracanceroustissues,epithelial dysplasias and normal tissuesThere is a higher Positive rate of Omi/HtrA2in oral squamouscarcinoma(74%,37/50) than in paracancerous tissue(34.0%,17/50)、 inepithelial dysplasias(38.0%,19/50)and in normal tissues(12.0%,6/50)(P <0.05).2The results of Omi/HtrA2are examined by RT-PCR in oral carcinomaspecimens，paracancerous tissues,epithelial dysplasias and normal tissuesThe gray value of Omi/HtrA2is12.5100±0.1454in well differentiatedoral squamous carcinoma, higher than10.1167±0.1475in poorly differentiatedoral squamous carcinoma,7.4678±0.2092in epithelial dysplasias,6.2314±0.2318in paracancerous tissue and1.2166±0.1651in normal tissues.There is a statistically significant among five groups (F=18417.435,p=0.000).The gray value of Omi/HtrA2in well differentiated oral squamous carcinomais higher than the other four groups,there is a statistically significant amongthem(P<0.05). According to the RT-PCR results, Omi/HtrA2has a Significantdifference in squamous carcinoma, epithelial dysplasias,paracancerous tissueand normal tissues, it showed an diminishing trend, which is Consistentedwith the results of immunohistochemistry.3The ralations between expression of Omi/HtrA2and PathologicalBy the statistic analysis, we obtained a result that immunohistochemistryand RT-PCR indicated expression of Omi/HtrA2correlated to the malignantdegree of cells,The expression of Omi/HtrA2in well differentiated oralsquamous carcinoma is highest(P<0.05).4The ralations between expression of Omi/HtrA2and clinical characteristicsin oral squamous carcinomaBy the statistic analysis, we obtained a result that immunohistochemistryindicated no correlation between expression of Omi/HtrA2and gender、age orClinical stages(P>0.05). Conclusion：1The expressions of Omi/HtrA2in oral squamous cell carcinoma tissues ishiger than in paracancerous tissues and normal tissues.There is a statisticallysignificant among them(P<0.05).2The expression of Omi/HtrA2has a positive correlation to thePathological,Omi/HtrA2in well differentiated oral squamous carcinoma ishigher than the other four groups.There is a statistically significant amongthem(P<0.05).3The expression of Omi/HtrA2in epithelial dysplasias is higher in normaltissues(P<0.05) but less than oral squamous carcinoma (P<0.05)so we canconjecture that the Omi/HtrA2play an important role in epithelial dysplasias.