| Objective:To investigate the expression of VEGF-C,VEFG-D and STAT3 in human gastric carcinoma tissues, adjacent gastric normal tissue, gastric cancer cell lines and gastric normal cell line. To explore the potential mechanism of VEGF-C and VEGF-D regulated by STAT3 contribution to the lymphangiogenesis. Methods:We detected the VEGF-C,VEGF-D and STAT3 expression in gastric cancer tissues, adjacent gastric normal tissues, gastric cancer cell lines and gastric normal cell line. Survival analysis and relationship analysis were performed. STAT3 si RNA transfection and genome microarray were applied to demonstrate whether the expression of VEGF-C and VEGF-D was mediated by the STAT3 in gastric cancer. Results: 1. 38.3% patients(41/107) showed positive expression of VEGF-C protein in tumor tissues, while 20.6%(22/1070 showed positive expression in adjacent gastric no-tumor tissues. However, VEGF-D, STAT3 and phosphorylation STAT3, they were 43.9% vs 14.0, 62.6% vs 8.4% and 38.3% vs 1.9%. The results show there were significant differences of VEGF-C, VEGF-D, STAT-3 and p STAT3 protein expression between gastric cancer tissues and adjacent gastric normal tissues(p VEGF-C=0.009, p VEGF-D=0.031, p STAT3=0.035, p p-STAT-3=0.014). 2. Univariate analysis showed VEGF-C, VEGF-D, STAT3 and p-STAT3 protein positive expression, sex, number of lymph node metastasis and extent of metastatic lymph nodes(p<0.05) have significant relationships with overall survival. P-STAT3 protein expression and number of lymph node were recognized as independent factors of overall survival after the multivariate analysis by using Kaplan-Meier. 3. The relative m RNA expression value of VEGF-C and VEGF-D in gastric cancer tissues were significantly much higher than those in adjacent gastric normal tissues(2.26±0.83 VS 0.40±0.12, p VEGF-C =0.007; 1.26±0.55 VS 0.75±0.29, p VEGF-D =0.041). Meanwhile, the relative m RNA expression value of STAT-3 in gastic cacner tissues were significantly much higher than those in adjacent gastric normal tissues(3.15±1.32 VS 1.13±0.64, p STAT3 =0.004). 4. STAT3 m RNA expression was much higher in AGS、SUN-1、KATO-III、BGC-823、MGC-803、SGC-7901 and HGC-27 gastric cancer cell lines than those in GES-1 normal cell line(p AGS =0.014, p SUN =0.020, p KATO-III =0.026, p BGC-823 =0.023, p MGC-803 =0.019, p SGC-7901 =0.018, p HGC-27 =0.014). At the sametime, VEGF-D m RNA expression was also much higher in AGS、SUN-1、KATO-III、BGC-823、MGC-803、SGC-7901 and HGC-27 gastric cancer cell line than those in GES-1 normal cell line(p AGS =0.012, p SUN =0.025, p KATO-III =0.017, p BGC-823 =0.015, p MGC-803 =0.009, p SGC-7901 =0.014, p HGC-27 =0.007). However, VEGF-C m RNA expression was not much higher in AGS、SUN-1、KATO-III、BGC-823、MGC-803、SGC-7901 and HGC-27 gastric cancer cell line than those in GES-1 normal cell line(p AGS =0.064, p SUN =0.435, p KATO-III =0.108, p BGC-823 =0.058, p MGC-803 =0.235, p SGC-7901 =0.083, p HGC-27 =0.072). 5. In the Immunohistochemistry result’s analysis, the number of lymphon node was significantly associated with VEGF-C, VEGF-D, STAT3 and p STAT3 proterin expression in GC tissues(p VEGF-C=0.062、p VEGF-D= 0.034、p STAT3 =0.021、pp-STAT3=0.004). Furtermore, STAT3 protein expression was significantly associated with VEGF-C and VEGF-D protein expression(p<0.05).While, p-STAT3 protein positive expression was only significantly associated with VEGF-D protein expression(p<0.05). In the m RNA relative expression result’s analysis, we also demonstrate that N stage was also significantly associated with VEGF-C, VEGF-D and STAT3 m RNA relative expression in gastric cancer tissues. Furthermore analysis, we found STAT3 m RNA relative expression was significantly associated with VEGF-D m RNA relative expression. 6. With STAT3 si RNA transfection, we demonstrated that VEGF-C and VEGF-D protein expression level decreased 20% to 30% and 70% to 80% compared with control group respectively. With the RNA microarray analyses, the 5295 genes expression was significant decreases in the HGC-27 cell with transfection of STAT3 si RNA. VEGF-C and VEGF-D were demonstrated to be down-regulated respectively. The relative RNA expression of VEGF-C decreased only 1.036-fold, while the VEGF-D significantly decreased 5.568-fold in the HGC-27 cell after STAT3 si RNA transfection compared with control group. Conclusion:1. The VEGF-C, VEGF-D and STAT3 protein expression and m RNA relative level in gastric cancer tissues were significantly higher than those in adjacent gastric normal tissues.2. Sex, N stage, and extent of metastatic lymph nodes, VEGF-C, VEGF-D, STAT and p-STAT3 protein expression were associated with the overall survival of GC. N stage and p-STAT3 protein expression were identified as the independent factors.3. VEGF-C, VEGF-D and STAT3 m RNA expression were much higher in AGS、SUN-1、KATO-III、BGC-823、MGC-803、SGC-7901 and HGC-27 gastric cancer cell lines than those in GES-1 normal cell line.4. The number of Lymph node was significantly associated with VEGF-C, VEGF-D, STAT3 and p-STAT3 protein and m RNA expression in GC tissues. Meanwhile, VEGF-C and VEGF-D protein and m RNA expression were also significantly associated with STAT3.5. With STAT3 si RNA transfection, VEGF-C and VEGF-D were down-regulated in protein and m RNA expression, especially the decrease of VEGF-D expression. STAT3 may be contribute to the lymph node metastasis by promoting lymphangiogenesis via up-regulation expression of VEGF-D... |
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