Silencing pancreatic adenocarcinoma up-regulated factor increases the sensitivity of pancreatic cancer cell line to gemcitabine

Abstract

Objective To observe the influence on the sensitivity of pancreatic cancer cell line BxPC-3 to gemcitabine of silencing PAUF gene. Methods BxPC-3 cells, which overexpress PAUF, was stably transfected with PAUF-shCtrl and PAUF-shRNA to establish BxPC-3_shCtrl and BxPC-3_shPAUF cells as control and experiment group. Then the mRNA and protein expression level of PAUF in these two cell lines were detected by RT-PCR and western blot, respectively. The growth inhibition rates of these two cell lines treated with different concentrations of gemcitabine (0, 3.1, 6.25, 12.5, 25, 50, 100, 200 nmol/L) were detected by MTT. Apoptosis rates in the cells treated with different concentrations of gemcitabine (0, 75, 100 nmol/L) were then observed by flow cytometry. Results The relative PAUF mRNA expression level in BxPC-3_shCtrl and BxPC-3 cells were 1.00±0.06 and 0.83±0.07, which were significantly higher than that in BxPC-3_shPAUF cells (0.25±0.02; both P<0.05). The relative PAUF protein expression level in BxPC-3_shCtrl and BxPC-3 cells were 0.89±0.07 and 0.95±0.04, which were significantly higher than that in BxPC-3_shPAUF cells (0.31±0.03; both P<0.05). The IC50 value of gemcitabine to BxPC-3_shCtrl cell was (22.88±2.43) nmol/L, which was significantly higher than that of BxPC-3_shPAUF cells [(1.06±0.02) nmol/L; P<0.05]; apoptosis rate of BxPC-3_shPAUF cells treated by gemcitabine increased faster than that of BxPC-3_shCtrl cells. Conclusion PAUF silencing could greatly enhance the sensitivity of BxPC-3 cells to gemcitabine. Key words: Pancreatic cancer cell line; Pancreatic adenocarcinoma up-regulated factor; Gemcitabine; Drug sensitization