| Objective:Head and neck cancer is the sixth most common types ofcancer in the world, among which the squamous cell carcinoma is the mostcommon type. Chemotherapy is one of the main methods for the treatment ofhead and neck squamous cell carcinomas. Neoadjuvant chemotherapy wasnamed by Frei in1982, and was made a preliminary report on its clinic effectby Skarin in1989. Then it was applied in the treatment of head and necksquamous cell carcinomas. And people has been exploring how to improve thecontrol and survival rate of the head and neck squamous cell carcinomas byneoadjuvant chemotherapy. With the further research of tumor formationmechanism, people gradually realized that the sensitivity of tumor cells toapoptosis is the key factor that decides the effect of chemotherapy. As thediscovery and further investigation of Omi/HtrA2, more and more studiespoint that Omi/HtrA2can induce cell apoptosis, and enhance the sensitivity ofcells to chemotherapy drugs. Meanwhile it confirmed that tumor may besuccessfully prevented by the intervention of Omi/HtrA2. We detect theexpression of Omi/HtrA2in head and neck squamous carcinoma tissues beforand after the intervention of nedaplatin, which is one of the neoadjuvantchemotherapy drugs, through the application of RT-PCR andimmunohistochemical technique, so as to provide theoretical basis for theapplication of neoadjuvant chemotherapy.Furthermore, we hope to find a waywhich could improve the control and survival rate of the head and necksquamous cell carcinomas.Methods：Specimens were collected from45patients with head and necksquamous cell carcinomas from March2012to August2013. All patientswere received a incisional biopsy, and got a clear diagnosis by pathology, then were given preoperative chemotherapy induced by nedaplatin before surgery.The specimens are divided into two groups immediately after they are cut off.One is immobilized in4%formaldehyde, then embed it by Paraffin, and theother is stored in-80℃.1Detecting the expression of Omi/HtrA2in the head and neck squamouscell carcinomas tissue and the para-carcinoma tissue before and after theintervention of nedaplatin through immunohistochemical technique.Make the wax blocks of each group to dewax to water, and operateaccording to the Maxvision2/HRP kit instructions, then observe under theelectron microscope.2Detecting the expression of Omi/HtrA2in the head and neck squamouscell carcinomas tissue and the para-carcinoma tissue before and after theintervention of nedaplatin through RT-PCR.The total RNA was extracted through the method of Trizol after groundthe tissue. And the exitracted RNA was identified by agarose gelelectrophoresis and defined quantitatively by UV spectrophotometer. Then theextracted RNA was compounded into the first chain of cDNA according to theinstructions of reverse transcriptase. Amplificate the purpose gene with usingthe β-acton as the gapdh. We used the gel imaging system for observation andimage acquisition.3Results criteria3.1ImmunohistochemistryThe possitive expression of Omi/HrA2staining was found Browngranules in cytoplasm, which was evaluated by Immunohistochemicalsemi-quantitative method.3.2Semi-quantitative analysis of gene expressionWe scanned the gray of GAPHD and objective products in order to gainthe relative expression,then analysised the results by Quantity One software.4Statistical MethodsWe uesed the statistical package SPSS13.0to analysis the data with the useing of chi-square test, repetitive measure analysis of variance and SNK twocomparison test method.(P<0.05for the difference was statisticallysignificant.)Results：1The results of Immunohistochemistry about the expressions of Omi/HtrA2in each group through.The results of Immunohistochemistry: Before the intervention ofnedaplatin, the expressions of the head and neck squamous cell carcinomastissue and the para-carcinoma tissue were64.4%（29/45）,13.3%（6/45), whichwere statistically significant after chi-square testing (P<0.05). After theintervention of nedaplatin, the expressions of the head and neck squamous cellcarcinomas tissue and the para-carcinoma tissue were80%(36/45),42.2%(19/45), which were statistically significant after chi-square testing (P<0.05).2The results of Immunohistochemistry about the expressions of Omi/HtrA2in different pathological grading.The results of Immunohistochemistry: Before the intervention ofnedaplatin, the expressions of Omi/HtrA2in well differentiated squamouscarcinoma tissues, poorly differentiated squamous carcinoma tissues, andparacancerous tissues were66.7%(24/36),55.6%(5/9),13.3%(6/45), whichwere statistically significant after SNK two comparison testing (P<0.05).After the intervention of nedaplatin, the expressions of Omi/HtrA2in welldifferentiated squamous carcinoma tissues, poorly differentiated squamouscarcinoma tissues, and paracancerous tissues were88.9%(32/36),66.7%(6/9),42.2%(19/45), which were statistically significant after SNK two comparisontesting(P<0.05).3The results of Immunohistochemistry about the expressions of Omi/HtrA2in each group before and after the intervention of nedaplatin.The results of Immunohistochemistry: The expressions of Omi/HtrA2inwell differentiated squamous carcinoma tissues before and after the intervention of nedaplatin were66.7%(24/36),88.9%(32/36). The expressionsof Omi/HtrA2in poorly differentiated squamous carcinoma tissues beforeand after the intervention of nedaplatin were55.6%(5/9),66.7%(6/9). Theexpressions of Omi/HtrA2in paracancerous tissues before and after theintervention of nedaplatin were13.3%(6/45),42.2%(19/45). The expression ofOmi/HtrA2genes in the three groups was not only statistically significant (P<0.05) within the groups before and after the intervention, but also between thegroups. According to the SNK two comparison testing, the expression ofOmi/HtrA2in the well differentiated squamous carcinoma tissues was higherthan it in the poorly differentiated squamous carcinoma tissues andparacancerous tissues, and was statistically significant (P<0.05).4The results of RT-PCR about the expressions of Omi/HtrA2in each groupthrough.The results of RT-PCR: Before the intervention of nedaplatin, the grayvalue of Omi/HtrA2in the head and neck squamous cell carcinomas tissueand the para-carcinoma tissue were0.7236±0.1353,0.3761+0.1526,whichwere statistically significant after chi-square testing (P<0.05). After theintervention of nedaplatin, the gray value of Omi/HtrA2in the head and necksquamous cell carcinomas tissue and the para-carcinoma tissue were1.2032±0.1450,0.4350+0.1172,which were statistically significant afterchi-square testing (P<0.05).5The results of RT-PCR about the expressions of Omi/HtrA2in differentpathological grading.The results of RT-PCR: Before the intervention of nedaplatin, the grayvalue of Omi/HtrA2in well differentiated squamous carcinoma tissues, poorlydifferentiated squamous carcinoma tissues, and paracancerous tissues were0.8126+0.1249,0.4627+0.1123,0.3761+0.1526, which were statisticallysignificant after SNK two comparison testing (P<0.05). After the interventionof nedaplatin, the gray value of Omi/HtrA2in well differentiated squamouscarcinoma tissues, poorly differentiated squamous carcinoma tissues, andparacancerous tissues were1.3553+0.1614,0.6718+0.1421,0.4350+0.1172, which were statistically significant after SNK two comparison testing(P<0.05).6The results of RT-PCR about the expressions of Omi/HtrA2in each groupbefore and after the intervention of nedaplatin.The results of RT-PCR: The expression of Omi/HtrA2in squamoucarcinoma tissues after the intervention of nedaplatin was significantly higherthan before the intervention. The gray value of Omi/HtrA2in the welldifferentiated squamous carcinoma tissues before and after the interventionof nedaplatin were0.8126+0.1249,1.3553+0.1614. The gray value ofOmi/HtrA2in the poorly differentiated squamous carcinoma tissues beforeand after the intervention of nedaplatin were0.4627+0.1123,0.6718+0.1421.The gray value of Omi/HtrA2in the paracancerous tissues before and afterthe intervention of nedaplatin were0.3761+0.1526,0.4350+0.1172. Theexpression of Omi/HtrA2genes in the three groups was not only statisticallysignificant (P<0.05) within the groups before and after the intervention, butalso between the groups. According to the SNK two comparison testing, theexpression of Omi/HtrA2in the well differentiated squamous carcinomatissues was higher than it in the poorly differentiated squamous carcinomatissues and paracancerous tissues, and was statistically significant (P<0.05).Conclusion：1The expressions of Omi/HtrA2in head and neck squamous cell carcinomatissues is higer than in paracancerous tissues. There is a statistically significantamong them(P<0.05).2The expression of Omi/HtrA2has a positive correlation to the Pathological.Omi/HtrA2in well differentiated squamous carcinoma is higher than thepoorly differentiated squamous carcinoma tissues and paracancerous tissues.There is a statistically significant among them(P<0.05).3The expression of Omi/HtrA2in the tissues after the intervention ofnedaplatin was higher than it before the intervention(P<0.05). We infer thatnedaplatin may promote the apoptosis by changing the expression ofOmi/HtrA2, as well as high expression of Omi/HtrA2may enhance the ability to kill cancer of nedaplatin. |
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