Gene Microarrays In Screening Out Apoptosis-related Genes Of Gemcitabine-mediated Apoptosis On Human Pancreatic Carcinoma Panc-1 Cells

Objective Pancreatic carcinoma is one of the most common and poorest prognosis abdomen malignant tumor with a low exairesis rate. Gemcitabine can obviously improve the prognosis of patients with advanced stage pancreatic carcinoma, but the apoptotic mechanism is unknown. This research is to screen out the different apoptosis-related genes after the pancreatic carcinoma cells have been cultured with Gemcitabine. The real time fluorescent PCR was used to testify the key apoptotic gene.Method The pancreatic cell line Panc-1 was cultured by DMEM. MTT colorimetric assay was used to detect the IC₅₀ of gemcitabine. FCM was used to detect the apoptotic rate of Panc-1 cells induced by gemcitabine. The technique of microarray was used to screen out apoptosis-related genes on human pancreatic carcinoma Panc-1 cells (gemcitabine-induced). ASK1 gene was quantitated relatively by the real time fluorescent PCR.Result The IC₅₀ of gemcitabine was 50.33μg/ml. After the cells were cultured with Gemcitabine for 48hr, the apoptotic rate rose significantly (P<0.01) . Microarray screened out overexpressed apoptosis-related genes including CyclinB1, ASK1, AKT2. On the other hand , it also screened out lowexpressed apoptosis-related genes including IRF-4, Scotin, 14-3-3gamma, PDE, GRPS, TID1, AFP, STAT5, Sox-4, c-Rel, NDSO, htt, CatD, gD. It displayed that the apoptotic pathway was caspase-independent ASK1 pathway. The real time...