Jansen Marc, Treutner Karl-Heinz, Schmitz Britta, Otto Jens, Jansen Petra Lynen, Neuss S, Schumpelick Volker
Department of Surgery, University Clinic, Pauwelsstr, 30, 52057 Aachen, Germany.
BMC Gastroenterol. 2004 Dec 29;4:33. doi: 10.1186/1471-230X-4-33.
Nidation of floating tumour cells initiates peritoneal carcinosis and limits prognosis of gastro-intestinal tumours. Adhesion of tumour cells to extracellular matrix components is a pivotal step in developing peritoneal dissemination of intraabdominal malignancies. Since phospholipids efficaciously prevented peritoneal adhesion formation in numerous animal studies we investigated their capacity to reduce adhesions of gastric cancer cells to extracellular matrix components (ECM).
Human gastric cancer cells (NUGC-4, Japanese Cancer Research Resources Bank, Tokyo, Japan) were used in this study. Microtiter plates were coated with collagen IV (coll), laminin (ln) and fibronectin (fn). Non-specific protein binding of the coated wells was blocked by adding 1% (w/v) BSA (4 degrees C, 12 h) and rinsing the wells with Hepes buffer. 50.000 tumour cells in 100 microl medium were seeded into each well. Beside the controls, phospholipids were added in concentrations of 0.05, 0.1, 0.5, 0.75 and 1.0/100 microl medium. After an incubation interval of 30 min, attached cells were fixed and stained with 0.1% (w/v) crystal violet. The dye was resuspended with 50 microl of 0.2% (v/v) Triton X-100 per well and colour yields were then measured by an ELISA reader at 590 nm. Optical density (OD) showed a linear relationship to the amount of cells and was corrected for dying of BSA/polystyrene without cells.
The attachment of gastric cancer cells to collagen IV, laminin, and fibronectin could be significantly reduced up to 53% by phospholipid concentrations of 0.5 mg/100 microl and higher.
These results, within the scope of additional experimental studies on mice and rats which showed a significant reduction of peritoneal carcinosis, demonstrated the capacity of phospholipids in controlling abdominal nidation of tumour cells to ECM components. Lipid emulsions may be a beneficial adjunct in surgery of gastrointestinal malignancies.
漂浮的肿瘤细胞着床引发腹膜癌，限制胃肠道肿瘤的预后。肿瘤细胞与细胞外基质成分的黏附是腹内恶性肿瘤发生腹膜播散的关键步骤。鉴于在众多动物研究中磷脂能有效预防腹膜粘连形成，我们研究了其降低胃癌细胞与细胞外基质成分（ECM）黏附的能力。
本研究使用人胃癌细胞（NUGC - 4，日本癌症研究资源库，东京，日本）。微量滴定板用IV型胶原（coll）、层粘连蛋白（ln）和纤连蛋白（fn）包被。通过加入1%（w/v）牛血清白蛋白（4℃，12小时）并用Hepes缓冲液冲洗孔来阻断包被孔的非特异性蛋白结合。将100微升培养基中50000个肿瘤细胞接种到每个孔中。除了对照组，以0.05、0.1、0.5、0.75和1.0/100微升培养基的浓度加入磷脂。孵育30分钟后，固定贴壁细胞并用0.1%（w/v）结晶紫染色。染料用每孔50微升0.2%（v/v）Triton X - 100重悬，然后用酶标仪在590nm处测量颜色产量。光密度（OD）与细胞数量呈线性关系，并针对无细胞的BSA/聚苯乙烯染色进行校正。
磷脂浓度为0.5mg/100微升及更高时，胃癌细胞与IV型胶原、层粘连蛋白和纤连蛋白的黏附可显著降低达53%。
这些结果，结合在小鼠和大鼠上进行的其他实验研究（显示腹膜癌显著减少），证明了磷脂控制肿瘤细胞在腹部着床于ECM成分的能力。脂质乳剂可能是胃肠道恶性肿瘤手术中的有益辅助剂。
