摘要
目的观察血管内皮生长因子C（VEGF-C）在人胰腺癌组织及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。方法采用免疫组织化学染色法检测15例人胰腺癌原发灶和转移淋巴结组织中VEGF—C的表达差异。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，应用逆转录聚合酶链反应（RT．PCR）、酶联免疫吸附实验（ELISA）、流式细胞术、原位末端标记法（TUNEL）进一步检测VEGF—C的表达差异，并通过VEGF—C反义寡核苷酸（ASODN）体内外转染抑制其表达，研究对淋巴结转移胰腺癌细胞凋亡的影响。结果人胰腺癌淋巴结转移组织中VEGF—C的表达水平明显高于原发组织（8．6±3．4比4．6±2．8，P〈0．05）；而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[mRNA：0．87±0．11比0．61±0．15，蛋白：（1682±157）pg／ml比（1404±128）pg／ml，均P〈0．05]。体内外转染VEGF—CASODN抑制其表达后，空白对照组、错义核苷酸组、ASODN组淋巴结转移胰腺癌细胞的凋亡率均显著提高[（2．8±1．0）％，（5．0±2．1）％，（13．2±2．2）％，均P〈0．01]，而原发灶胰腺癌细胞无明显影响[（1．8±0．5）％，（2．0±0．7）％，（4．4±1．0）％，均P〉0．05]。结论在人胰腺癌组织及动物模型中，淋巴结转移灶胰腺癌细胞VEGF—C的表达均明显高于原发灶，并且其表达下凋能特异性促进淋巴结转移胰腺癌细胞的凋亡。
Objective To investigate the expression of vascular endothelial growth factor C （ VEGF- C） and the effect of antisense oligonucleotide （ASODN） upon lymph node metastasis of pancreatic cancer cell. Methods We selected 15 cases of human pancreatic cancer and detected the expression of VEGF-C in primary tumor and lymph node metastasis tissues with immunohistochemistry. Meanwhile, the spontaneous lymphatic metastasis model in nude mice was established with orthotopic 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 upon the apoptosis of pancreatic cancer cell derived from primary tumor and spontaneous lymphatic metastasis were detected by reverse transcription polymerase chain reaction （RT-PCR）, enzyme linked immunosorbent assay （ELISA）, flow cytometer and terminal deoxylnucleotidyl transferase mediated-dUTP nick end labeling （TUNEL）. Results In tissues of human pancreatic cancer, the values of VEGF-C on lymph nodes metastasis were more higher than primary tumor （ P 〈 0. 05 ）. About the expression of VEGF-C on pancreatic cancer cell derived from spontaneous lymphatic metastasis and primary tumor in nude mice model, the mRNA levels of VEGF-C were 0. 87±0. 11 and 0. 61 ±0. 15 respectively, the VEGF-C levels in culture supernatants were （1682 ± 157）pg/ml and （1404 ± 128）pg/ml. The expression of VEGF-C on pancreatic cancer cells derived from lymphatic metastasis were also more higher than primary tumor （ P 〈 0. 05 ） . In vitro and vivo, transfection of VEGF-C ASODN decreased the expression of VEGF-C in pancreatic cancer cell. In control group, scramble-sense oligonucleotide （SODN） group and ASODN group, the apoptosis rates of pancreatic cancer cells derived from lymph node metastasis were （2. 8 ± 1.0 ） % , （ 5.0 ± 2. 1 ） % , （ 13.2 ± 2. 2 ） % respectively in vitro, and were （ 1.8 ± 0. 5 ） %, （ 2. 0 ± 0. 7 ） % , （4. 4 ± 1. 0） % respectively in vivo, the apoptosis was increased significantly after transfection of VEGF-C ASODN （all P 〈0. 01 ）. But pancreatic cancer cells derived from primary tumor were not effeeted （ all P 〉 0. 05 ）. Conclusion In human pancreatic cancer and nude mice model, the expression of VEGF-C on lymphatic metastasis was higher than primary tumor. The apoptosis of pancreatic cancer cells derived from spontaneous lymphatic metastasis were promoted by transfeetion of VEGF-C ASODN specially.
出处
《中华医学杂志》
CAS
CSCD
北大核心
2009年第34期2386-2390,共5页
National Medical Journal of China
关键词
胰腺肿瘤
血管内皮生长因子C
淋巴转移
细胞凋亡
原位种植
Pancreatic neoplasms
Vascular endothelial growth factor C
Lymphatic metastasis
Apoptosis
Orthotopic implantation
