The Expression Of Hpa And Mmp-9Protein In Non-small Cell Lung Cancer And Its Clinical Significance

Objective:To study preliminarily the effects of HPA and MMP-9in the occurrence, invasion and metastasis of non-small cell lung cancer (NSCLC) by detecting the expression of them in non-small cell lung cancer (NSCLC). Methods:Detect HPA and MMP-9expression level in120cases of non-small cell lung cancer (NSCLC) tissues from pathology department of Luzhou Medical College (60lung squamous carcinoma,60lung adenocarcinoma) and30cases of normal lung tissue by immunohistochemical SP method. NSCLC tissues were obtained from120NSCLC patients, without any radiotherapy or chemotherapy before surgeries and other invasive operation.30controls are normal lung tissues. The tissue samples are treated with the10%formalin-fixation, paraffin-embedding, and4μm serial sections conventionally. After dewaxing and hydrating paraffin sections conventionally, they were rinsed in distilled water, then, immerse them in phosphate buffer saline (PBS) for5minutes. Antigens are repaired by high pressure and heating. Then incubate them with3%hydrogen peroxide methanol solution for10minutes, to eliminate endogenous peroxidase activity. Block the non-specific staining caused by the cross-reaction, with normal goat serum. Primary antibody incubation (the dilution ratio of HPA polyclonal rabbit-anti-human antibody is1:50; and the dilution ratio of MMP-9rabbit-anti-human polyclonal antibody is1:100). Wash them with PBS, then replace primary antibody by PBS for negative control. Secondary antibody inucubation with biotin-labeled antibody: wash them with PBS, then, incubate by Avidin-Biotin Peroxidase Complex working solution(S-A/HRP). After PBS washing, DAB staining, Mayer hematoxylin redyeing, acidification, bluing, gradient dehydrated, transparentizing, mounting, observe them by microscope. Pay attention to control the background. Positive cells are the cells that there are claybank granules or masses in their cytoplasm or nucleus. The most positive signals of HPA are in the nucleus, and a few of them are in the cytoplasm, while positive signals of MMP-9are usually found in cytoplasm. The slices are divided into four grades by their proportion of positive cells and depth of staining:-,+,++,+++. The sign "-" means non-staining or staining cells account for less than5%of all cells in a slice, and the sign "+" means light-staining or staining cells account for between5%to25%. The sign "++" means moderate-staining or staining cells account for between26%to50%, and the sign "+++" means deep-staining or staining cells account for more than50%. Results:(1) The expression of HPA and MMP-9in NSCLC is significantly higher than that in normal lung tissues (P<0.05),(2) The difference of expression of HPA in lung squamous carcinoma and adenocarcinoma has no significance (P>0.05),while the expression of MMP-9in lung adenocarcinoma is significantly higher than lung squamous carcinoma (P<0.05).(3)HPA and MMP-9are expressing significantly higher in group which with low differentiation than well and moderately differentiation(P<0.05).3groups based on TNM stage have different expression of HPA and MMP-9(P<0.05).By Multiple comparisons,the expression of HPA and MMP-9in group of II-III stage(TNM stage) are significantly higher than that in group of I stage(P<0.05), and the group of IV stage has higher expression of HPA and MMP-9than group of I stage and group of II-III stage (P<0.05)。(4)In NSCLC, the expression of HPA and MMP-9are positively correlated with each other, and it is statistically significant (r=0.416,P <0.005) Conclusion:(1) HPA and MMP-9are both probably take part in carcinogenic process and formation of NSCLC.(2) HPA and MMP-9are both participating in the progress, invasion and metastasis of NSCLC. It implies that over-expression of HPA and MMP-9may promote the formation and progress of NSCLC.(3) HPA and MMP-9perhaps cooperate with each other in the formation, progress, invasion and metastasis of NSCLC.