低渗HEPES缓冲溶液对腹腔内游离肝癌细胞杀伤作用的实验研究

Experimental Study on The Killing Effect of Hypotonic Solution for Intraperitoneal Free Cancer Cells of Hepatocellular Carcinoma

目的探讨肝癌细胞在低渗溶液中的存活状态,为临床应用温热蒸馏水杀灭腹腔游离肝癌细胞提供理论依据。方法对体外培养的肝癌细胞株Hep3B分别应用渗透压为148 m Osmol/kg和90 m Osmol/kg的羟乙基哌嗪乙硫磺酸(HEPES)缓冲溶液及0 m Osmol/kg的蒸馏水作用后,运用MTT法及流式细胞仪观察其活性及状态。结果 148 m Osmol/kg及90 m Osmol/kg渗透压的HEPES缓冲溶液对肝癌细胞株Hep3B的杀伤作用不明显,作用30 min后仍有较强的活性;0 m Osmol/kg渗透压的蒸馏水对肝癌细胞有一定的杀伤作用,且随作用时间的延长其杀伤效果增强,作用10 min后才可杀灭肿瘤细胞。结论在肝癌手术中应用温热蒸馏水灌洗需要持续作用10 min以上方可起到杀灭腹腔内游离癌细胞的作用,进而预防术后腹膜种植转移的发生。

Objective To investigate the survival of liver cancer cells in hypotonic solution, and provide a theoretical basis for the clinical application of warm distilled water to kill free intraperitoneal liver cancer cells. Methods The hepatocellular carcinoma cell line Hep3B were cultured by using different osmotic HEPES buffered solution （148 mOsmol/kg, 90 mOsmol/kg, and 0 mOsmol/kg） in vitro, and then the activity and status of the cells were observed by using the MTT assay and flow cytometry. Results One hundred and forty-eight mOsmol/kg and 90 mOsmol/kg osmotic pressure solution had no obvious killing effect for hepatocellular carcinoma cell line Hep3B, the cells still had strong activity after 30 rain. The 0 mOsmol/kg osmotic pressure distilled water had particular killing effect for hepatocellular carcinoma cells, with the prolongation of time and the killing effect was enhanced. It needs 10 min to destruct free cancer cells. Conclusion Application of warm distilled water lavage in liver cancer surgery requires the continuing role of 10 rain to kill the intraperitoneal free cancer cells, thereby preventing the occurrence of postoperative peritoneal metastasis.