Pross Matthias, Lippert Hans, Misselwitz Frank, Nestler Gerd, Krüger Sabine, Langer Harald, Halangk Walter, Schulz Hans-Ulrich
Department of Surgery, Otto-von-Guericke University of Magdeburg, Leipziger Strasse 44, D-39120 Magdeburg, Germany.
Thromb Res. 2003 Jun 1;110(4):215-20. doi: 10.1016/s0049-3848(03)00296-2.
Metastases, adhesion and invasion of tumor cells involve a cascade of complex phenomena, which potentially can be affected by glycosaminoglycans. We studied the influence of a low-molecular-weight heparin, reviparin, on the intraabdominal tumor growth in rats undergoing laparoscopy. We also studied cytotoxicity, anti-adhesive, and anti-invasive effects of reviparin in vitro using adenocarcinoma cells CC531.
In vitro assays: Adhesion of 1 x 10(5) CC531 adenocarcinoma cells onto microtiter plates coated with 10 microg/ml collagen type I or 10 microg/ml Matrigel was studied in the presence of 0.55; 1.10 and 2.76 mg/ml reviparin, and compared to saline. The cytotoxicity of 1 x 10(4) adenocarcinoma cells was studied in a similar assay. Transwell dual chambers with polycarbonate filters coated with 100 microg/cm2 Matrigel were used to investigate the effect of 0.55; 1.10 and 2.76 mg/ml reviparin on the invasion of 1 x 10(5) adenocarcinoma cells/ml. In vivo experiments: CC531 adenocarcinoma cells (5 x 10(6) cells/ml) were intraperitoneally applied to Wistar Albino Glaxo rats (n=150, Harlan, Germany) with a median weight of 278 g. The rats were divided into 15 groups with 10 animals in each group, underwent laparoscopy, and 1 ml saline containing 0, 0.5, 2.0, 4.0, and 10 mg reviparin per kg b.w. was introduced for intraperitoneal lavage or s.c. After 21 days the animals underwent an autopsy, and the tumor weight was determined.
In vitro experiments: We found a highly significant inhibition of tumor cell adhesion and invasion (p<0.001) by all reviparin concentrations used in the assays. There was no effect of reviparin on the viability of cells in the cytoxicity assay. In vivo experiments: We found that application of 4.0 and 10.0 mg/kg b.w., but not 0.5 or 2.0 mg/kg b.w. significantly (p<0.01) decreased the tumor mass compared to controls, receiving only saline. This effect was most pronounced after the combined i.p. and s.c. application, whereas after a sole i.p. application, only the highest dose of 10 mg/kg b.w. caused a significant inhibition of tumor growth.
Low-molecular-weight heparin, reviparin, given in combination of i.p. lavage and s.c. injections, significantly diminishes intraabdominal tumor growth of CC531 adenocarcinoma cells in rats undergoing laparoscopy. This may offer additional therapeutic options for patients undergoing laparoscopic cancer surgery.
肿瘤细胞的转移、黏附和侵袭涉及一系列复杂现象，而糖胺聚糖可能会对这些现象产生潜在影响。我们研究了低分子量肝素瑞肝素对接受腹腔镜检查的大鼠腹腔内肿瘤生长的影响。我们还在体外使用腺癌细胞CC531研究了瑞肝素的细胞毒性、抗黏附及抗侵袭作用。
体外实验：在存在0.55、1.10和2.76mg/ml瑞肝素的情况下，研究1×10⁵个CC531腺癌细胞黏附到包被有10μg/mlⅠ型胶原或10μg/ml基质胶的微量滴定板上的情况，并与生理盐水组进行比较。在类似实验中研究1×10⁴个腺癌细胞的细胞毒性。使用包被有100μg/cm²基质胶的聚碳酸酯滤膜的Transwell双室，研究0.55、1.10和2.76mg/ml瑞肝素对1×10⁵个/ml腺癌细胞侵袭的影响。体内实验：将CC531腺癌细胞（5×10⁶个细胞/ml）腹腔注射给体重中位数为278g的Wistar白化Glaxo大鼠（n = 150，德国Harlan公司）。将大鼠分为15组，每组10只动物，接受腹腔镜检查，每千克体重腹腔灌洗或皮下注射含0、0.5、2.0、4.0和10mg瑞肝素的1ml生理盐水。21天后对动物进行尸检并测定肿瘤重量。
体外实验：我们发现实验中使用的所有瑞肝素浓度均对肿瘤细胞黏附和侵袭有高度显著的抑制作用（p < 0.001）。在细胞毒性实验中，瑞肝素对细胞活力没有影响。体内实验：我们发现，与仅接受生理盐水的对照组相比，每千克体重注射4.0和10.0mg，但不是0.5或2.0mg，能显著（p < 0.01）降低肿瘤质量。这种作用在腹腔内和皮下联合给药后最为明显，而仅腹腔内给药时，只有最高剂量10mg/kg体重能显著抑制肿瘤生长。
低分子量肝素瑞肝素，通过腹腔灌洗和皮下注射联合给药，能显著减少接受腹腔镜检查的大鼠腹腔内CC531腺癌细胞的肿瘤生长。这可能为接受腹腔镜癌症手术的患者提供额外的治疗选择。
