title: Cisplatin combined with hyperthermia kills HepG2 cells in intraoperative blood salvage but preserves the function of erythrocytes.
abstract: The safe use of intraoperative blood salvage (IBS) in cancer surgery remains controversial. Here, we investigated the killing effect of cisplatin combined with hyperthermia on human hepatocarcinoma (HepG2) cells and erythrocytes from IBS in vitro. HepG2 cells were mixed with concentrated erythrocytes and pretreated with cisplatin (50, 100, and 200 μg/ml) alone at 37 °C for 60 min and cisplatin (25, 50, 100, and 200 μg/ml) combined with hyperthermia at 42 °C for 60 min. After pretreatment, the cell viability, colony formation and DNA metabolism in HepG2 and the Na(+)-K(+)-ATPase activity, 2,3-diphosphoglycerate (2,3-DPG) concentration, free hemoglobin (Hb) level, osmotic fragility, membrane phosphatidylserine externalization, and blood gas variables in erythrocytes were determined. Pretreatment with cisplatin (50, 100, and 200 μg/ml) combined with hyperthermia (42 °C) for 60 min significantly decreased HepG2 cell viability, and completely inhibited colony formation and DNA metabolism when the HepG2 cell concentration was 5×10(4) ml(-1) in the erythrocyte (P<0.01). Erythrocytic Na(+)-K(+)-ATPase activity, 2,3-DPG level, phosphatidylserine externalization, and extra-erythrocytic free Hb were significantly altered by hyperthermia plus high concentrations of cisplatin (100 and 200 μg/ml) (P<0.05), but not by hyperthermia plus 50 μg/ml cisplatin (P>0.05). In conclusion, pretreatment with cisplatin (50 μg/ml) combined with hyperthermia (42 °C) for 60 min effectively eliminated HepG2 cells from IBS but did not significantly affect erythrocytes in vitro.
authors.affiliation: Department of Anesthesiology, the Second Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou 310009, China; Jiangsu Province Key Laboratory of Anesthesiology, Xuzhou Medical College, Xuzhou 221004, China.
authors.forename: Jin-ting
authors.initials: JT
authors.lastname: Yang
authors.forename: Li-hui
authors.initials: LH
authors.lastname: Tang
authors.forename: Yun-qing
authors.initials: YQ
authors.lastname: Liu
authors.forename: Yin
authors.initials: Y
authors.lastname: Wang
authors.forename: Lie-ju
authors.initials: LJ
authors.lastname: Wang
authors.forename: Feng-jiang
authors.initials: FJ
authors.lastname: Zhang
authors.forename: Min
authors.initials: M
authors.lastname: Yan
mesh_terms: D019794:2,3-Diphosphoglycerate / Q000737:chemistry; D000328:Adult; D000368:Aged; D000970:Antineoplastic Agents / Q000627:therapeutic use*; D002470:Cell Survival; D002945:Cisplatin / Q000627:therapeutic use*; D003131:Combined Modality Therapy; D004305:Dose-Response Relationship, Drug; D004354:Drug Screening Assays, Antitumor; D004912:Erythrocytes / Q000187:drug effects*; D005260:Female; D006454:Hemoglobins / Q000737:chemistry; D056945:Hep G2 Cells; D006801:Humans; D006979:Hyperthermia, Induced; D008297:Male; D008875:Middle Aged; D057725:Operative Blood Salvage; D009995:Osmosis; D010718:Phosphatidylserines / Q000737:chemistry; D010743:Phospholipids / Q000737:chemistry; D000254:Sodium-Potassium-Exchanging ATPase / Q000737:chemistry
chemical_list: D000970:Antineoplastic Agents; D006454:Hemoglobins; D010718:Phosphatidylserines; D010743:Phospholipids; D019794:2,3-Diphosphoglycerate; D000254:Sodium-Potassium-Exchanging ATPase; D002945:Cisplatin
keywords: Cisplatin; Erythrocytes; HepG2 cells; Hyperthermia; Intraoperative blood salvage