| ObjectiveTo study the distribution of LPPCN((linearly patterned programmed cell necrosis) by morphological observation and indicate the clinical and pathological significance in malignant melanoma,and to analyze the mechanism of LPPCN and the relationship between LPPCN.and Vasculogenic Mimicry (VM) and angiogenesis.Methods1. To identify the existence and distribution of LPPCN by morphological observation and investigate clinical pathological significance of LPPCN.Seventy melanoma cases undergoing surgery with definite pathologic diagnosis and integrated follow-up document in Tianjin Cancer Hospital from 1996 to 2007 were collected in this study.The H&E-stained sections of every specimen was observed again and checked the pathologic diagnosis by salty pathological doctor.The LPPCN's morpho logyical feature and clinical pathological significance were observed.2. To analyze the mechenism of LPPCN and the relationship between LPPCN and neovascularization.To detect the expression of ClassⅢPI3K and analyze the relation between LPPCN and autophagy.The expression of MMP2,MMP9,TIMP-1,CathD, CD105,TGFβ1,p-paxillin,Ⅷ-R Ag,and D2-40 were examined by immunohist ochemistry.VMD of these samples were evaluated by immunohistochemical and histochemical double staining of CD31 and PAS.The relationship between LPPCN and VM,angiogenesis,lymphangiogenesis were analyzed.3. To examine the different expression of ClassⅢPI3K,TGFβ1 between LPPCN-positive group and the negative one by Real-time PCR,and analyze the relation between ClassⅢPI3K,TGFβ1 and LPPCN.Thirty Fresh and refrigerant cases of Tissue Bank Department in Tianjin Medical University Cancer Hospital were collected from 2005 to now.LPPCN was observed by the H&E-stained frozen section.All the cases were divided into LPPCN-positive group and the negative one.The expression of ClassⅢPI3K and TGFβ1 were detected by Real-time PCR between the two groups,so that the mechanism of LPPCN and the relation between LPPCN and angiogenesis were analyzed from mRNA.Results 1. The morphological feature and clinical pathological significance of LPPCN in melanoma.Of the 70 cases of melanoma,54.29%(38/70) were recognized to have LPPCN.The tumor cells undergoing LPPCN were a cluster of spindle cells with condensed cytoplasm and dark nuclei which were distributed in patterns of lines and networks in the H&E-stained sections. LPPCN was correlated with the size of tumor,mitosis figure count and Breslow depth,but there was no relationship between LPPCN and gender,age,site, tumor embolus,lymph node metastasis and recurrence, distant metastasis. Kaplan-Meier survival analysis showed the survival rate of the patients with LPPCN was lower than that of patients without LPPCN(P=0.046).2. The result of immunohistochemistry stainingIn melanoma,the expressions of ClassⅢPI3K,the microvessel density (MVD) labeled by CD105,TGFβ1,Ⅷ-R Ag-MVD,VMD and MMP2 were higher in LPPCN-positive group than that in negative one,and there were significant difference (P<0.05);However,there were no significant difference for the expressions of the lymphvessel density(LVD) labeled by D2-40,MMP9 and CathD between the two groups (P>0.05);the expression of ClassⅢPI3K and TGFβ1 in the regions of LPPCN was obviously higher than that in circumambient tumor cells (P<0.05).The expression of TGFβ1 was positively correlated with CD105-MVD;there were positive correlations between CD105-MVD,Ⅷ-R Ag-MVD,VMD and LPPCN.There was low or no expression of p-paxillin in all the melanoma tissues.P-paxillin also can express on the endothelial cell of the blood vessel.The expression of MMP2 and MMP9 in VM-positive group was higher than that in negative one (P<0.05);and the expression of TIMP1 in VM-positive group was lower than that in negative one (P<0.05);there was no significant difference for the expression of CathD between the two groups.The expression of LVD labeled by D2-40 in the group with lymph node metastasis was higher than that in the negative one (P<0.05)3. The result of Real-time PCRIn malignant melanoma,the expression of ClassⅢPI3K,TGFβ1mRNA in LPPCN-positive group was higher than that in the negative one.This result was conformed to the result of immunohistochemistry.There was significant difference only for the expression of TGFβ1mRNA between the two groups (P<0.05) Conclusions1. LPPCN and VM exists in malignant melanoma,and the patients with LPPCN and VM had a bad prognosis.2. LPPCN may be a phenomenon of autophagy from tumour cells,and was associated with VM and angiogenesis. Some tumor cells undergoing LPPCN can establish a spacial foundation for neovascularization.3. The study didn't demonstrate the correlation between LPPCN and lymphangiogene sis.D2-40,as a new marker of lymphatic endothelial cells,was correlated with lymph node metastasis in melanoma.D2-40 can help to evaluate the prognosis of patients.
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