热双蒸水联用右旋糖酐-40杀灭瘤细胞及抗瘤细胞粘附的体外实验研究

In vitro studies on prevention of intraperitoneal neoplasm seeding by local application of hyperthermic distilled water combined with dextran-40

目的 评价热双蒸水 (DDW)灭瘤及右旋糖酐 40 (Dextran 40 )抗瘤细胞粘附的体外效果。方法 分别用温热DDW ,温热DDP(阳性对照 )及NS(阴性对照 ) ,浸泡B16 F10 瘤细胞 10min及 2 0min ,2 4hr后行活细胞计数。在处理过的培养板孔内分别加Dextran 40及PBS(对照组 ) ,接种B16 F10 细胞 ,16hr后收集贴壁活细胞计数。结果 浸泡 10min时与阴性对照组比 ,温热DDW组和阳性对照组活细胞数均显著减少 (P<0 .0 1) ,后者减少更明显 (P <0 .0 5 ) ;但浸泡 2 0min后 ,两组均无细胞存活 (无差异 )。与对照组比 ,Dextran 40可部分抑制B16 F10 细胞对胶化培养板孔壁的粘附 (P <0 .0 1)。结论 温热DDW浸泡 2 0min能完全杀灭体外培养的B16 F10 细胞。Dextran 40可部分抑制B16 F10 细胞对胶化培养板孔壁的粘附。

Objective To evaluate the efficacy of hyperthermic distilled water combined with dextran 40 as propylaxis against intraperitoneal tumor seeding in vitro. Methods Twelve wells among 24-well tissue culture plates seeded with 1 4×10 4 200μl B 16 F 10 melanoma cells per well were divided into three groups and each well was added with 45℃ distilled water or 45℃ DDP(0.56mg/100mlNS) or saline respectively. Ten or twenty minutes later, the experimental agents were aspirated and the cells were incubated for 24 hours, then harvested. The viable cells were counted and the number were compared usingt test or t'test. Eight wells among 24-well tissue culture plates were gelatnized and divided into two groups. Each well was added with 200μl 10% dextran 40 or PBS respectively then inoculated with 0.7×10 4/100μl B 16 F 10 cells. The plates were incubated for 16 hours, then harvested and analyzed in the same manner metioned above. Results There were no viable tumor cells in the tissue culture wells exposed for 20 min to any tumoricidal agent, whereas confluent growth occured in all wells exposed to control medium. While B 16 F 10 cells were exposed for 10 min to 45℃ DDW or 45℃ DDP, the number of viable cells both decreased significantly(P<0.01, by comparing with the control group respectively.) Comparison of the efficacy of dextran 40 group vs. PBS(control) group showed that dextran 40 partially blocked attachment of the B 16 F 10 cells to gelatinized wells(P<0.01). Conclusions In vitro test showed that B 16 F 10 cells were completely killed when exposed to 45℃ DDW for 20 min and their attachment to gelatinized wells was partially inhibited by dextran 40.