| ObjectiveLaryngeal papilloma is a benign laryngeal tumor.There are several distinctive clinical manifestation in adult laryngeal papilloma,such as more generate single,canceration easily and is one of precancerous conditions with highly rate of canceration.Because influence of biopsy and pathologist's experience,it is not ideal to early diagnosis canceration of PL.Malignant tumor has a procedure that is a multiple factors and steps.Phosphorylation of proteins on serine or threonine residues preceding praline(Ser/Thr—Pro)is an important signaling mechanism controlling cell differentiation and proliferation.Pinlis peptidyl-prolyl cis/trans isomerases that regulate caryocinesia of cell.Pin1 can specifically identify and isomerizes the phosphorylated Ser/Thr—Pro bonds in certain proteins,thereby modulating their functions.CyclinD1 is a multifunctional protein that plays an important role in the transduction of Ras signals.Interestingly,overexpression of Pin1 can contribute to the aberrant expression of CyclinD1 in the nucleus,which enhances the abnormality generation of cell..Recent research indicates that Pin1 and CyclinD1 are overexpressed in many human cancers and play an important role in oncogenesis.So we investigated the expression of Pin1,CyclinD1 and the relationship of their expression in laryngeal papilloma, laryngeal papilloma with mild,severe atypical hyperplasia and laryngeal carcinoma in order to evaluate their possible roles in the canceration of LP.Methods1.Patient selectionTissue samples of 39 cases of adult laryngeal papilloma,27 cases of with mild and severe atypical hyperplasia and 16 cases of laryngeal carcinoma(n=82)and normal laryngeal tissues(n=10)were obtained from surgical and biopsy patients of the Sheng Jing hospital of China Medical University between 1989 and 2007.2.Immunohistochemistry10 normal tissues formalin-fixed and paraffin embedded specimens was available for this study.All paraffin sections were cut a 4μm thickness,after repairing antigen and blocking nonspecific reactivity,the sections were indubated with anti-Pin1 polyclonal and anti-CclinD1 monoclonal antibody and then with biotin-conjugated second antibody.Positive signals were detected the SP method with SP kit.Sections were also counterstained with hematoxylin.3.Result determination①Pin1:The normal staining was in the nucleus;the overexpressed staining was in the nucleus and/or cytoplasm.(A)The staining intensity was scorded as negative (0);weak(1);morderate(2),and strong(3);(B)The number of cells stained from 5 selected fields was graded as follows:0%(0);1%～33%(1);34%～66%(2);≥67%(3).A×B,Grades≥3 showed its overexpression.②CyclinD1:The overexpressed staining was in the nucleus and/or cytoplasm.Less than 10%of cells stained showed negative,and over 10%of cells stained showed positive.4.Statistical methodsAll the data were analyzed with SPSS for Windows 11.0 software.We used x~2 or Fisher's exact test to analyze the correlation expressions of Pin1 and CyclinD1 between the group and the other.We used spearman rank correlation to analyzed the dependability expressions of Pin1 and CyclinD1 in all groups.P value of less than 0.05 or 0.0125 was accepted as statistically significant.Results1.Expression of Pin1 in normal tissues and pathological tissuesPin1 was almost no expressed in normal tissues(0/10).Otherwise,Pin1 was overexpressed in LP(12.82%),LP with mild and severe atypical hyperplasia(40.74%)and laryngeal careinoma(81.25%),and localized in nucleus and/or cytoplasm.The dist ribution of Pin1 is increased gradually in the course of from laryngeal normal epithelial tissue to laryngeal carcinoma(p＜0.05);There was significantly difference among all groups(p＜0.0125). 2.Expression of CyclinD1 in normal tissues and pathological tissuesCyclinD1 was almost no expressed in normal tissues(0/10).Otherwise,CyclinD1 was overexpressed in LP(17.95%),LP with mild and severe atypical hyperplasia(33.33%)and laryngeal carcinoma(75%),and localized in nucleus and/or cytoplasm.The dist ribution of CyclinD1 is increased gradually in the course of from laryngeal normal epithelial tissue to laryngeal carcinoma(p＜0.05);There was significantly difference among rest groups(p＜0.0125),but between LP and LP with mild,severe atypical hyperplasia(p＞0.0125)in the expression of CyclinD1.3.Correlation expression of Pin1 and CyclinD1In 7 cases of CyclinD1 overexpressed,there was 4 cases of Pin1 overexpressed in LP.There was significantly direct correlation in the expression of Pin1 and Cyc-linD1 (r_s=0.620,P＜0.001)analyzed by spearman rank correlation test.In addition,for 24 cases of Pin1 overexpressed,there was 17 cases of CyclinD1overexpressed in LP with mild,severe atypical hyperplasia and laryngeal carcinoma(r_s=0.495,P＜0.005).Conclusion(1)The Pin1 and CyclinD1 were expressed in LP,LP with mild,severe atypical hyperplasia and laryngeal carcinoma that may produce a marked effect in laryngeal papilloma's malignant change.(2)There was significantly direct correlation in the expression of Pin1 and Cycli-nD1.Pin1 up-regulate the expressions of cyclinD1,and participate possibly it's malignant change.
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