Effects On Lymphangiogenesis Of Lung Cancer With Antisense Oligonucleotides Of Vascular Endothelial Growth Factor C

Objective:Vascular endothelial growth factor C(VEGF-C) recently has been identified as a new member of the VEGF family and is considered as the single specific lymphangiogenic factor.VEGF-C and its receptor VEGFR-2/VEGFR-3 are the key players in the molecular regulation of lymphangiogenesis.Many experiments have highlighted the role of VEGF-C in turmor biology. Some studies have recently shown that VEGF-C mediated tumor lymphangiogenesis promotes cancer metasatisis including nodal metastasis. The aim of this article is to investigate the effects on lymphangiogenesis of orthotopic implantation of lung cancer in nude mice with antisense oligonucleotides of VEGF-C.Methods: Establishment of the model in nude mice with orthotopic implantation for the human lung cancer cell line A549.Thirty nude mice were randomized into three groups : PBS control group(PBS),sense oligonucleotides control group(SODN) and antisense oligonucleotides group(AODN). Inoculation of human lung adenocarcinoma cell line A549, after three days, with PBS, SODN and AODN were treated with subcutaneous injection, weekly observation of lung cancer grown in nude mice subcutaneous tumor growth rate and characteristics. Four weeks after treatment, all mice were sacrificed and tumors were excised for routine histological examination, body weight,HE staining。After examination were completed,the expression of VEGF-C mRNA, protein and lymphatic microvessel density(LMVD) of lung cancer were detected by RT-PCR ,Western Blot and immunohistochemistry.Results: The oligonucleotide intervention after injection, the tumor volume ofeach group within a week was no significant difference in growth rate. Two weeks after the tumor volume was observed AODN group (38.2±1.6) significantly smaller than the other two groups (P<0.01)。AODN group 2 weeks after the growth was slow, or even stop growth, but no significant tumor regression was observed situation. SODN group and PBS group is still growing tumor growth, but SODN tumor volume was slower than that of PBS group of tumor growth in a little slow, but no significant difference between (P> 0.05)。After 4 weeks observation AODN Tumor volume (58±11.7) compared with the previous two groups statistically significant difference (P<0.01), cultivation of tumor weight(0.56±0.11)compared with the other two groups statistically significant difference(P<0.05). After treatments, the expression of VEGF-C mRNA in AODN group(4.61±1.57)was inhibited significantly (P<0.05). The LMVD in AODN group(3.2±1.7) was decreased significantly, there were significant differences compared with control groups(P<0.01)。Conclusion: Liposome-mediated VEGF-C antisense oligonucleotides can reduce lung cancer grown in nude mice subcutaneous tumor growth rate, while significantly reducing the expression of VEGF-C mRNA and protein,and its micro-lymphangiogenesis has strong inhibition,it is possible to prevent metastasis of lung cancer tumor cells.