Expression Of MMP-7 In NSCLC And Effect Of MMP-7 On Vessel Endothelial Cell

Objectives: 1. To examine the expression of MMP-7(matrilysin) and CD34 in NSCLC (non-small cell lung cancer), and analyze the relation between the MMP-7 and MVD (microvessel density). 2. To explore the proliferation and inhibition of HUVEC (human umbilical vein endothelial cell) and A549 (lung adenocarcinoma cell line) on the condition of rMMP-7 (recombinant matrilysin) and LIPO- MAON (antisense oligonucleotide of matrilysin by liposome transfection)Methods: 1.Examination of the expression of MMP-7 and CD34 in 57 NSCLC by S-P immunohistochemistry assay. 2. Cell primary culture of HUVEC. 3. Cell infection of LIPO- MAON by liposomes. 4. Examination of the proliferation and inhibition of HUVEC and A549 on the condition of rMMP-7 and LIPO- MAON by MTT assay. 5. Examination of the expression of HUVEC and A549 on the condition of rMMP-7 and LIPO- MAON by Semiquantitative RT-PCR assays. Results: 1.The result for examination of 7 normal lung tissue and 57 NSCLC by Immunohistochemistry assay showed: the positive rate of MMP-7 was 14.3%(1/7) in 7 normal lung tissue, 82.5% (47/57) in 57 NSCLC, 82.9%(29/35) in adenocarcinoma, and 81.8%(18/22) in squamous carcinoma, there was no differentiation between adenocarcinoma and squamous carcinoma; 78.3%(18/23) in well-moderately differentiated NSCLC, and85.3%(29/34) in poorly differentiated NSCLC, there was a differentiation between normal lung tissue, well-moderately differentiated NSCLC and poorly differentiated NSCLC(P<0.05); there was a differentiation between differentiation of NSCLC and expression of MMP-7(P<0.05); Average microvessel density (MVD) was 15.1 in normal lung tissue, 39.4 in well-moderately differentiated NSCLC, 51.5 in poorly differentiated NSCLC, the average MVD was 45.8 in NSCLC. There was no differentiation on MVD between adenocarcinoma and squamous carcinoma; there was a differentiation on MVD between the normal lung tissue, well-moderately, and poorly differentiated NSCLC (P<0.05). Accompanied with the increase of malignant degree of NSCLC, the MVD was added, there was a differentiation between expression of MMP-7 and MVD (P<0.05).2. The result on promotion and inhibition to HUVEC and A549 by MTT assay in case of rMMP-7 and LIPO-MAON revealed: 0.25μg, 0.5μg/ml rMMP-7 can't promote the proliferation of HUVEC, 0.75μg, 1.0μg/ml rMMP-7 can do it; 2.5nmol-10nmol/ml LIPO-MAON can't inhibit the proliferation of HUVEC; 0.25μg/ml rMMP-7 can't promote the proliferation of A549, 0.5μg, 0.75μg, 1.0μg/ml rMMP-7 can do it; 2.5nmol, 5nmol/ml LIPO–MAON can't inhibit the proliferation of A549, 7.5nmol, 10nmol/ml LIPO-MAON can do it obviously. In a word, high concentration MMP-7 have corresponding promotion to the proliferation of blood vessel endothelial cell and lung cancer cell, and any concentration LIPO-MAON can't inhibit the proliferation of blood vessel endothelial cell, but high concentration LIPO-MAON can inhibit the proliferation of lung cancer cell.3. The result on MMP-7-mRNA to HUVEC and A549 by RT-PCR assay in case of rMMP-7and LIPO-MAON displayed: HUVEC didn't express MMP-7-mRNA in any case; rMMP-7 enhanced the expression of MMP-7-mRNA in A549, and LIPO-MAON decreased the expression of MMP-7-mRNA in A549 (P<0.05).Conclusion: There was a higher expression of MMP-7 in NSCLC (82.5%), which correlated to the cell differentiation and MVD, not to the pathological type; there was a expression of MMP-7-mRNA in lung adenocarcinoma cell, not in vessel endothelial cell; High concentration rMMP-7 can promote the increase of vessel endothelial cell and lung adenocarcinoma cell; LIPO–MAON inhibited the increase of vessel endothelial cell inefficiently, nevertheless inhibited lung adenocarcinoma cell efficaciously.