VEGF-C在胰腺癌中的表达及对淋巴结转移影响的体内外实验研究

Expression of VEGF-C on pancreatic cancer and the effect on lymph node metastasis

目的研究VEGF-C在胰腺癌裸鼠原位种植瘤模型中的表达特点,及其表达抑制后对胰腺癌细胞淋巴结转移的影响。方法建立人胰腺癌细胞株PANC-1裸鼠原位种植瘤模型,原代培养原发和淋巴结转移灶中胰腺癌细胞,应用RT-PCR、ELISA进一步检测VEGF-C的表达差异,并通过VEGF-C反义寡核苷酸体内外转染抑制其表达,应用流式细胞术及TUNEL等方法体内外研究对淋巴结转移胰腺癌细胞凋亡的影响。结果胰腺癌裸鼠原位种植瘤模型中淋巴结转移胰腺癌细胞VEGF-C的表达水平也显著高于原发灶细胞(P<0.05)。体内外转染VEGF-C反义寡核苷酸抑制其表达后,淋巴结转移胰腺癌细胞的凋亡率均显著提高(P<0.01),而原发灶胰腺癌细胞的凋亡率无明显影响(P>0.05)。结论在胰腺癌裸鼠原位种植瘤模型中,淋巴结转移灶胰腺癌细胞VEGF-C的表达均明显高于原发灶,并且其表达下调能特异性促进淋巴结转移胰腺癌细胞的凋亡,这可能是VEGF-C在胰腺癌淋巴结转移中的新机制。

[Objective] To investigate the expression of vascular endothelial growth factor C （VEGF-C） and the effect of antisense oligonucleotide （ASODN） on lymph node metastasis of. pancreatic cancer cell. [Methods] Establishment of the spontaneous lymphatic metastasis model in nude mice with orthotopie implantation for the human pancreatic cancer cell line PANC-1, isolation and culture of primary tumor and spontaneous lymphatic metastasis. The effect of VEGF-C special ASODN on apoptosis of pancreatic cancer cell derived from primary tumor and spontaneous lymphatic metastasis were detected by reverse transcription polynlerase chain reaction （RT-PCR）, enzyme linked immunosorbent assay （ELISA）, flow cytometer, terminal deoxylnucleotidyl transferase mediated-dMTP nick end labeling （TUNEL） in vitro and vivo. [Results] The mRNA level of VEGF-C on pancreatic cancer cell derived from spontaneous lymphatic metastasis was also more higher than primary tumor in nude model （P 〈0.05）. In vitro and vivo, transfection of VEGF-C ASODN decreased the expression of VEGF-C in pancreatic cancer cell. But, only the apoptosis of pancreatic cancer cells derived from spontaneous lymphatic metastasis was increased significantly （P 〈0.01）, pancreatic cancer cells derived from primary tumor were not effected （P 〉0.05）. The volume of tumor, rates and parameter of lymph node metastasis all were not statistical differentiation （P 〉0.05）. [Conclusion] The expression of VEGF-C on pancreatic cancer derived from different organ was discrepancy. The apoptosis of pancreatic cancer cells derived from spontaneous lymphatic metastasis were promoted by transfeetion of VEGF-C ASODN specially, but no effect on primary tumor. It is new mechanism of lymph node metastasis in pancreatic cancer.