| IntroductionLung cancer is one of the most common malignity carcinoma. In recent years,the incidence rate and the death rate are increasing worldwild including our country, especially the city mortality from lung cancer resulting from air pollution or cigaratte smoking. Non - small cell lung cancer accounts for 80% ~ 85% of all lung tumors.The incidence of lung cancer is a complicate process in which many genes take part and go through a lot of stage developments. All kinds carcinogenic factors change genes of cell including ILK. ILK is defined and cloned by HanniganGE in 1996. It is a 59 KD serine/threonine proterin kinase which contains ankyrin repeat domain. It is a focal adhesion protein that has multiple domains. ILK has many directly biochemical activities. It has been implicated in the regulation of signal transduction of cell and the functions on survival, proliferation, differentiation and migration of cell. Every little change of activity or expression of ILK may elicit the change of signal transduction and cell cycle subsequently. These may result in human tumor or some disease which have relationship with cell reproduction and the intra - extracellular matrix.By now , there s no study about the expression of ILK in lung cancer and the study of ILK in other human tumor are not coincidence. We now investigate expression of intergrin - linked kinase( ELK) and the correlation between ILK expression and clinicopathological characteristics and prognosis of non - small cell lung cancer( NSCLC) to evaluate the significance of ILK in NSCLC.Materials and methodsTissue samplesParaffin blocks were obtained from 58 patients with primary NSCLC who underwent curative operation at the Iiaoning Tumor Hospital from November 1999 to February 2000. 10 NSCLC speciments were obtained from patients with surgical excision at the First Hospital affiliated to China Medical University in 2002. Other 33 NSCLC speciments were obtained from patients with surgical excision at the Shenyang junqu 202 Hospital. ILK protein were detected by immunohisto-chemistry in 101 NSCLCs. Western blot was used to detected ELK protein in 16 fresh speciments.ImmunohistochemistryDetect the ILK expression by immunohistochemistry S - P method.. The staining intensity was scored as negative (0) ; weak staining (1 + ); moderate staining (2 + ) ; strong staining (3 + ).Western blotRapidly homogenize tissuses (0. 125 cm3 ) in 5 volumes of lysis buffer (50mM Triscl PH7.4, 150mM Nacl, 0.1% SDS, 1% Triton-100, 1mM ED-TA PH 8.0, Aprotinin 12ul/10ml, PMSF 60ul/10ml). Centrifuge the homoge-nate (10000rpm,4C) for 30 minutes to pellet insoluble material. Electrophore-sis transfer proteins from gel to PVDF blocking incubation with primary antibody enzyme conjugate incubation substrate incubation. Scan the results to computer.Statistical analysisAll the data were analyzed with SPSS For WindowslO. 0 software. Survival analysis used Kaplan - Meier and Cox model; The relationship between expression of ILK protein and clinical pathological characters was analyzed with chi -squared test. The cut - off for statistical significance was defined as P <0.05.ResultsImmunostaining location and expression of ILK:Normal smooth muscular cell and fibroblat show strong staining, but the e-valuation of result dosen' t include them. Normal bronchial epithelium and type II alveolar cell show moderate staining; the cell of NSCLC show strong staining; type I alveolar cell and lymphocyte show negative staining.Correlation of protein expression of ILK with clinical pathological features;The strong staining of ILK in squmous cell cancer is52% , in adenocareino-ma the rate is 58% , theres no significance (P =0.433 >0.05). In squamous cell cancer , the strong staining of moderate and low differentiated cancer is higher than well differentiated samples (P =0.001 <0.05). There is no stasti-cal significance in different differentiated adenocarcinoma. The strong staining of ILK has no statically significant correlation with clinical grad... |
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