Effects Of Recombinant HIL-24Combined With Cisplatin On The Growth Of Endometrial Carcinoma Cell Line HEC-1B In Vitro

Objective:To explore the effective drug dosage for application in vitro through the combination use of recombinant human interleukin-24(rhIL-24) and cisplatin(DDP) for the inhibition of endometrial carcinoma cell line HEC-1B, so as to study the anti-tumour mechanism and provide theoretical guidance for the clinical combination use of rhIL-24and DDP for in the treatment of endometrical carcinoma.Methods:1.100ul of HEC-1B cells and100ul WI-38cells with a concentration of2×104cells/ml were added into a96-hole plate, and cultured in the CO2culture box in condition of37℃.5%CO2and saturated humidity. When complete adherent growth took place in both of the cell lines. rhIL-24solution with a concentration of80.100.120.150.200ng/ml and DDP solution with a concentration of5.10.25.50.75μg/ml were added separately for further culture of24h,48h and72h. CCK-8assay was used to measure the growth inhibition rate separately.2.100ul of HEC-1B cells and100ul WI-38cells with a concentration of2×104cells/ml were added into a96-hole plate, and cultured in the COt culture box in condition of37℃,5%CO2and saturated humidity. When complete adherent growth took place in both of the cell lines,120ng/ml of rhIL-24and8μg/ml of DDP were added for further culture of24h.48h and72h, CCK-8assay was used to measure the growth inhibition rate separately.3.100ul of HEC-1B cells and100ul WI-38cells with a concentration of2×104cells/ml were added into a50-ml flask, and cultured in the CO2culture box in condition of37℃.5%CO2and saturated humidity. When complete adherent growth took place in both of the cell lines.120ng/ml of rhIL-24,10μg/ml of DDP and the combination of120ng/ml of rhIL-24and8ug/ml of DDP were added for further culture of24h. cell apoptotic rate were measured by FCM separately.4.100ul of HEC-1B cells and100ul WI-38cells with a concentration of2×104cells/ml were added into a50-ml flask, and cultured in the CO2culture box in condition of37℃5%CO2and saturated humidity. When complete adherent growth took place in both of the cell lines,120ng/ml of rhIL-24.10μg/ml of DDP and the combination of120ng/ml of rhIL-24and10μg/ml of DDP were added for further culture of24h and48h. the morphological changes of apoptotic cells were observed under fluorescence microscopy separately.Results:1. The growth inhibition rate of120ng/ml of rhIL-24for HEC-1B cells is18.0%. the inhibition rate is lower when the concentration of rhIL-24-s is higher or lower; there is no apparent inhibition of WI-38cells for rhIL-24; the inhibition rate of DDP is concentration dependent.2. The growth inhibition rate of120ng/ml rhIL-24combined with10μg/ml DDP for HEC-1B cells was59.0%.88.2%and91.9%separately after being cultured for24h.48h and72h; the growth inhibition rate for WI-38cells was63.1%,88.7%and92.4%separately.3. For120ng/ml of rhIL-24and10μg/ml of DDP,after being cultured for24h. the apoptotic rate of HEC-1B cells was14.5%and26.8%separately by FCM analysis:and WI-38cells, was6.8%and17.1%separately.4. For the combination of120ng/ml of rhIL-24and10μg/ml of DDP. the apoptotic rate of HEC-1B cells was29.6%by FCM analysis; and WI-38cells was20.1%5. For120ng/ml of rhIL-24.10μg/ml of DDP and the combination of120ng/ml of rhIL-24and10μg/ml of DDP. the morphological changes of apoptotic cells were observed under fluorescence microscopy, and the results showed that:normal cells were not stained; the apoptotic cells were dyed grass green, and the nuclei was destroyed irregularly or agglutinated being beaded; the dead cells without apoptosis were stained in whole.Conclusion:1. Different concentrations of rhIL-24can inhibit endometrial carcinoma cells, and the inhibition effects were in a non-dose-dependent manner, with the appropriate inhibitory concentration of120ng/ml; different concentrations of DDP can inhibit endometrial carcinoma cells, and the inhibition effects were in a dose-dependent manner. DDP also had inhibition effects on normal cells.2. The solution of rhIL-24. DDP or the combination of both agents had inhibition effects on endometrial carcinoma cells, the inhibition rate of the combination use was higher than the one of singal use, and the combinaton use showed synergistic effects.3. The solution of rhIL-24. DDP or the combination of both agents had the induction of apoptosis of endometrial carcinoma cells, and rhIL-24inhibited the growth of tumor cells through the mechanism of apoptosis. there was no inhibtion effects on WI-38cells, which suggested that the anti-tunmor effects were through apoptosis.