| Backgrounds and AimsLung cancer is one of the best common malignant tumor in the world.It is a kind of disease that threats severely the health and safety of people.The disease incidence and death rate of lung cancer of every country in the world all offer obvious and adscendent tendency since half century.The report of WHO in 1997:The death rate of lung cancer is the first one cancers. The disease incidence of NSCLC occupies more than 70%.The five years survival rate of patients with NSCLC is average 16%.Among the total .The five years survival rate of patients whose tumours have partly metastasised is average 23%, but it is only 3% that patient's tumour has metastasised to distant place.The basic reason of cure failure and patient death is the metastasis.So revealing the mechanism of metastasis of cancer cell and predicting the potentiality of metastasis of cancer cell will conduce clinician to judge patient's condition,establish reasonable treatment plan and judge prognosis and conduce to mitigate patient's distress,extend patient's life span and raise quality of life.Studies completed disclose that the invasion and metastasis of lung cancer cell is a very complicated biology course that is controled by many factors.It relates to the malignant multiplication of lung cancer cell in the primary focus of infection cancer cell breaking away from the primary focus of infection,forming cancer embolia and migration in vessel,implantating in target organ and forming new metastases.The key is cancer cell breaking away from the primary focus of infection and how to adhere target organ.Cell adhesion participates the whole course and acts a important role.CAM and catenins mediate cell adhesion.CAM include Selectins,IGSF,Cadherins and so on.In the recent years , many scientises have paied close attention to Catenins and Cadherins.They have become the hot spot among the molecule mechanism of metastasis.Cadherins is a group of CAM depending on Ca2+,the main constituent of AJ structure and made up with a series of single strand transmembrane glycoprotein.They possess very important effect in the processes of cell adhesion and differentiation, especially the earlier period of differentiation.We have known there are more than 40 varietases Cadherin now. Study about N-cadherin is very poor, especially in NSCLC.There are several reports about it in foreign country,but only one in our country. Study discover N-cadherin acts a important role in the adhesion of tumour epithelium tissue cell,blood vessel smooth muscle cell and cutaneous cell.In the process of tumour formation, N-cadherin encourages blood vessel formation and the transference between epithelium tissue cell and stroma cell.It can activate MAPKERK system through FGF,then induce the expression of MMP-9 gene .These two factors profit tumour vessel formation and make tumour growth and metastasis easy.Catenins have four subtypes,α,β,γand P120-catenin.The reports about the relationship betweenγ-catenin and NSCLC are few. Cadherin/catenins cell adhesion complex could not maintain normal adhesion,if have not the participation ofγ-catenin.Only whenγ-catenin separates from cell adhesion complex,cell could infiltration and metastasis.After the tyrosine residue ofγ-catenin becomes phosphorylation,will effect the stability of Cadherin/catenins cell adhesion complex. All these could induce canceration. So studing the expression ofγ-catenin in NSCLC and the relationγ-catenin with metastasis is very important to diagnosis and therapy of NSCLC.According to the study state at present, the aim of this topic is to explore more deeply the expression and clinical significance of N- cadherin andγ-catenin in NSCLC and understand the relation between they and the genesis,development and metastasis of NSCLC,then approach in some extent the mechanisms of NSCLC's genesis, development and metastasis,in order to provid some Assistance to diagnosis and therapy of NSCLC.Study Object and Methods1.Study object: Samples were collected from patients with primary NSCLC who were operated in the first affiliated hospital of zhengzhou university from September in 2002 to October in 2005.The sum total was 61 examples. Among the total,42 examples were male and 19 examples were female.The age of patients were form 36 years to 81 years,and averaged 57.77 years.All patients were not taken radiotherapy and chemotherapy. The pathological type of all samples were witnessed by pathologists,36 examples were squamous carcinoma and 25 examples were adenocarcinoma.According to UICC standard of 1997,these samples were divided in:â… A stage 2 examples,â… B stage 12 examples,â…¡A stage 0 example,â…¡B stage 7 examples,â…¢A stage 9 examples,â…¢B stage 19 examples,â…£stage 12 examples. The thickness of sections was 4μm.in addition,20 examples normal lung tissue(the distances form cancer are all above 3 cm) by the side of cancer acted as comparison.2.Methods: S-P immunohistochemical method was used.ALL steps were strictly operated according to the directions of S-P kit.Made PBS take the place of the first antibody as negative comparison.The masculine comparison of N-cadherin was normal heart muscle.The masculine comparison ofγ-catenin was breast cancer tissue.The main steps were : (1)deparaffinage and hydration according to common practice and reparating antigen through boiling;(2)Using peroxydase blocking agent to block the activity of endogenous peroxydase;(3)nonimmune animal blood serum blocked non-specificity reflection;(4)dropwising mouse anti-human N-cadherin monoclonal antibody or 1:100γ-catenin polyclonal antibody;(5)dropwising the second antibody;(6)dropwising streptavidin-peroxydase praeparatum;(7)dyeing with DAB,then washing sufficiently with tap water;(8)afterstaining with hematoxylin,then dehydrating,mounting and testing under microscope.3.Assessing result(1)N-cadherin: counting 100 cells every visual field under high power lens(400 times). Counting 10 visual fields. According to literature,divide stain intension in three stages: no stain(1 stage) 1 point,Buff(2 stage) 2 point,brown(3 stage) 3 point; According to the percentage of the positive cells occuping cancer cells, the percentage≤25% 1 stage (point), 26%～75% 2 stage (point),≥75% 3 stage (point), the ultimum score of evry section is the cross product of two counting(1 point～9 point).Dividing it in two stage: masculine(+) >7 point, negative (-) <7 point. (2)γ-catenin: counting 100 cells every visual field under high power lens(400 times). Counting 10 visual fields, masculine (+):We could see brown drop in cell.The drop was stained uniformitily and the positive rate≥50%. negative (-):A11 cells were not stained or the positive rate≤50%.4.treatment in statistics: Using SPSS 10.0 and thex2test and exact probabilities in 2×2 table treats data. P<0.05Results1.The expression of N-cadherin andγ-catenin in NSCLC:The results of immunohistochemistry displayed that N-cadherin andγ-catenin located on cell envelope or cytoplasmand located perchanc nucleus.N-cadherin was negative in all tumor-adjacent normal tissues.the positive rate of N-cadherin expression in NSCLC tissues was 37.70%(23/61)(P<0.01).γ-catenin was negative in all tumor-adjacent normal tissues.the positive rate ofγ-catenin expression in NSCLC tissues was 72.13%(44/61)(P<0.01).2.The relation N-cadherin andγ-catenin with the clinical patho-character : The positive rate of N-cadherin expression (61.54%) in the poorly differentiated NSCLC tissues was significant higher than that(31.25 %) in the well and moderately differentiated NSCLC tissues (P<0.05). In addition , The expression rate of N-cadherin was related to the tumor stage( P<0.05). The expression rate of N-cadherin was not related to histologic type,having or no pleural effusion,lymph node metastasis,age,sex,tumor location and tumor size.The positive rate ofγ-catenin expression (84.21%) in the NSCLC tissues of having lymph node metastasis was significant higher than that(52.17 %) in the NSCLC tissues of no lymph node metastasis (P<0.05). In addition ,γ-catenin expression rate was related to the grade differentiation and tumor stage(all .P<0.05). The expression rate ofγ-catenin was not related to histologic type,having or no pleural effusion,age,sex,tumor location and tumor size.Conclusion1.The positive rate of N-cadherin andγ-catenin expression were significant higher than normal tissues.It revealed that they were close correlation and could become a new diagnosis marker of NSCLC.2.The expression rate of N-cadherin was related to the tumor stage and differentiation and was not related to histologic type,having or no pleural effusion,lymph node metastasis,age,sex,tumor location and tumor size.3. The expression rate ofγ-catenin was related to the tumor stage,differentiation and lymph node metastasis and was not related to histologic type,having or no pleural effusion,age,sex,tumor location and tumor size.4.The high expression of N-cadherin andγ-catenin could offer important proof to judge patient's condition and establish reasonable treatment plan. |
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