Over Expressions Of PTTG And VEGF-C In Non Small Cell Lung Cancer; The Efects Of VEGF-C Antisense Oligonucleotides On Apoptosis Of A549 Cells

BackgroundNon Small Cell Lung Cancer(NSCLC) is among the human main malignant tumors, which has higher invasion and bad prognosis , angiogenesis is required for tumor development, progression and metastasis.The theory about the important effects of angiogenesis to tumor development has already been accepted widely.However,we can find out that metastasisis of NSCLC is more likely induced by lymph metastasis.The study of lymph node metastasis is slowly for the short of definite lymphatic microvessel grows facor and symbol. VEGF-C is the only definite lymphatic vessel endothelial cel 1 grows facor be found up to now. The role of VEGF-C to Lymph angiogenesis of the tumor is noticed by many people.In recent years, we find that VEGF-C can enhance Tumor lymph angiogenesis by activating lymphatic vessel endothelial cell receptor VEGFR-3' tyrosine phosphorylation. Pituitary tumor transforming gene (PTTG) was an oncogene be found in 1997.The studies show that the over expressions of VEGF-C can be promoted by PTTG and VEGF-C plays an important role in lymph node metastasis. Section I Over expressions of PTTG and VEGF-C in Non-small Cell Lung Cancer and their clinical significancesObjective: To investigate lymphatic microvessel density and expressions of pituitary tumor transforming gene and vascular endothelial growth factor C in non small cell lung cancer, and explore their correlations to clinicopathologic features of NSCLC and the clinical significances.Methods: Real-time fluorescence quantitative PCR and Immunohistochemistry method were used to detect the expressions of PTTG,VEGF-C and LMVD in NSCLC. We also investigate the effects of PTTG and VEGF-C to the prognosis of patients with NSCLC by drawing Kaplan-Meier curves.Results: PTTG,VEGF-C and LMVD have higher expressions in NSCLC than in adjacent tissues and benign lesions(P<0.05).There were no significant differences among the expressions of PTTG,VEGF-C and age,sex,smoking,tumor diameter,histological type and differentiation(P>0.05). There were statistical relationships between their expressions and TNM stage,lymph node metastasis and prognosis(P<0.05).The Kaplan-Meier curves showed that patients with low or no expressions of PTTG or VEGF-C have longer survival time than those with high expressions.The expression of PTTG was positively correlated with VEGF-C expression and LMVD.Conclusion: Tumor lymph angiogenesis was enhanced by the overexpression of PTTG and VEGF-C.The expressions of PTTG and VEGF-C may be used as two useful markers to judge the biological behavior and prognosis of lung cancer.VEGF-C may be used as a new target of antilymphatic therapy for lung cancer. Section II The efects of VEGF-C antisense oligonucleotideson apoptosis of A549 cellsObjective: To study the effects of VEGF-C antisense oligonucleotides on apoptosis of A549 cells.Methods: After VEGF-C antisense oligonucleotides was transfected into A549 cells for 48 hours, growth appearance and morphology of the cells were observed with immunofluorescence microscope, agarose gel electrophoresis was performed for the DNA ladder and the percentage of apoptosis cells was analyzed by flow cytometric annexin V-FITC/PI. MTT technique was also used to observe the multiplication of different group cells.Results: After being acted by VEGF-C antisense oligonucleotides, the growth of A549 cells was inhibited, the nuclei of the cells shrinked and the apoptosis body was observed in the plasma of the cells. The DNA ladder was clearly detected in agarose gel electrophoresis, FCM analysis showed that the percentage of apoptosis cells in VEGF-C antisense oligonucleotides group was 28.21±0.75% .significantly higher than that in control group 4.37±0.48% and negative group 3.26±0.35%. The growth curve observed by MTT showed that multiplication was slower in VEGF-C antisense oligonucleotides group cells.Conclusion: VEGF-C antisense oligonucleotides could significently inhibit A549 cells, it causes the cells to apoptosis and inhibits A549 cells to proliferate. VEGF-C antisense oligonucleotides may become one of the new methods of targeting gene therapy to lung cancer.