The Effect Of Cisplatin Combined With Hyperthermia In Different Temperature On The Cell Growth And Metastasis Of Human Ovarian Cancer Cell Line Skov-3

Objective:To investigate the effect of cisplatin combined with hyperthermia in different temperature on the cell growth and metastasis of human ovarian cancer cell line skov-3.Metheds:Skov-3 cell was cultured in vitro. According to heating condition of hyperthermia, the cells were divided into eight groups randomly: 37℃group (control group),39℃group,40℃group,41℃group,42℃group,43℃group,44℃group,45℃group. Cisplatin concentration is 5μg/ml, the heat time is lasting 60 minutes.(1)Inverted phase contrast microscope was used to observe morphologicalchanges of the cells.(2)MTT assay was used to determine the inhibition rate of cell growth.(3)Flow cytometry(FCM) was used to analyse changes of cell cycle and apoptosis rate.(4)Immunohistochemical method was used to detect the expression of Heat Shock Protion 70.(5)Reverse transcription polymerase chain reaction(RT-RCR) technique was used to observe the expression of S100A4 mRNA.Results: (1)Under inverted phase contrast microscope, cells of 37℃group (control group) grew well and proliferated continually, while cells of the other groups grew slowly, have different morphological changes: the cells appeared round and bright, the mount of the cells decreased. Cells of 43℃group changed most obviously: hardly were there any normal cells and many round floating cells and plentiful cell fragments were observed. Cells of 44℃group and 45℃group changed uniform with 43℃group.(2)Compared with control group, the inhibition rate of cells in the other groups increased(P<0.05), the apoptosis rate increased significantly(P<0.01),while the expression of S100A4 mRNA decreased significantly (P<0.01);When the temperature reached 43℃,the inhibition rate and the apoptosis rate increased maxlmum, the expression of S100A4 mRNA decreased minimum. There were no differences during 43℃,44℃and 45℃groups(P>0.05).(3)Cell cycle analysis by FCM indicated that with increase of the propotion of cells in G0/G1 phase and decrease of S phase, the cell cycle could be blocked gradually at G0/G1 phase by hyperthermia(P<0.05), the differences during these groups were significant(P<0.01).(4) Compared with control group, the expression of HSP70 by immunohistochemical method in 39℃～42℃increased(P<0.05), which caused heat tolerance easily,but there were no differences during these four groups(P>0.05). When the temperature reached 43℃, the expression of HSP70 decreased gradually,there were no differences during 43℃,44℃and 45℃group(P>0.05).Conclusions: Cisplatin chemotherapy in different temperature could inhibit the growth and invasion of skov-3 cell in vitro,43℃group affects most apparently. Cisplatin chemotherapy in 43℃could decrease heat tolerance and improve effect of the treatment in ovarian cancer. 43℃may be the most suitable temperature in intraperitoneal hyperthermic chemotherapy(IPHC).