Hepatoblastoma Cell Line의 항암제 감수성 검사

Abstract

Background: Hepatoblastoma is the most frequent malignant hepatic tumor in children which has been treated with surgery, radiation and chemotherapy. To increase the cure rate, in vitro drug sensitivity test might be needed. Recently we established a hepatoblastoma cell line (KPH1). This cell line could be useful as a model in drug sensitivity studies of hepatoblastoma. Methods: KPH1, cells are continuously cultured with RPMI media (10% FCS) in a incubator in 5% CO2 at 37℃. Cells were exposured with adriamycin, vincristine and VP-16 for 1 hr at the concentration of 0.1, 1.0 and 10 ㎍/ml. Cell viability was observed using trypan blue dye exclusion method. Chemosensitivity test was assessed with the MTT assay. Ninety six-well microplate contained 100μl cell suspension with the drugs was incubated for 4 days. Then 50 yt MTT solution was added for 6 hours. The formazan crystals formed by living cells were dissolved with DMSO (150μ1). The optical density (OD) was measured with a microplate spectrophotometer at 565nm. The OD was linearly related to cell number. Cell line (KPH1, K562, Raji and EU1 ) survival and LD50 (drug concentration lethal to 50% of the cells) were calculated and cell cycle assay was done using flow cytometry. Results: Cell viability decreased due to higher concentration and longer exposure to the drugs. KPH1 cell line showed sensitivity to the drugs such as adriamycin, vincristine and VP-16 which are also clinically effective to the patients. Adriamycin had synchronizing effect of KPH1 cell line to S phase. Conclusion: KPH1 cell line could be used as an experimental model of cancer chemosensitivity test. Further study to evaluate the chemosensitivity with multiple drugs and tumor cells from patients will be needed.