Effect of gemcitabine on proliferation, apoptosis and polo-like kinase-1 expression of pancreatic cancer cell line AsPC-1

Abstract

Objective To investigate the effect of gemcitabine on proliferation,apoptosis and polo-like kinase-1 (PLK-1) expression of human pancreatic cancer cell line AsPC-1 cells.Methods AsPC-1 cells were treated with various concentrations of gemcitabine for different time.The effect of gemcitabine on survival of AsPC-1 cells was determined by methyl thiazol tetrazolium (MTT) assay.The effect of gemcitabine on apoptosis of AsPC-1 cells was mearsured by flow cytometry with various concentrations of 2 and 5 μmol/L,and at different time points (12,24,48,and 72 h).The expression levels of PLK-1 mRNA and protein were detected by semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting,respectively.Results Gemcitabine decreased the proliferation of pancreatic cancer cells (P ＜ 0.05),and the apoptosis rate was significantly increased (P ＜ 0.05).After treatment with gemcitabine,the expression levels of PLK-1 mRNA (control:0.70 ±0.15,0.65 ±0.27,0.72 ±0.13,0.68 ±0.30;2 μmol/L:0.88 ±0.58,0.95 ±0.53,2.36 ±0.57,3.53 ±0.89;5 μmol/L:0.89 ± 0.60,1.02 ± 0.32,3.95 ± 1.84,4.52 ± 2.25) and protein (control:0.82 ± 0.41,0.78 ± 0.46,1.01 ±0.05,0.88 ±0.25 ;2 μmol/L:0.89 ±0.28,1.61 ±0.88,1.66 ±0.18,3.28 ± 1.33 ;5 μmol/L:0.85 ± 0.36,1.89 ± 0.18,2.73 ± 0.78,4.32 ± 2.13) were also increased significantly in pancreatic cancer cells (P ＜ 0.05).Conclusion Gemcitabine can influence the proliferation and apoptosis of AsPC-1 cells probably by the expression changes of PLK-1. Key words: Pancreatic carcinoma; Gemcitabine ; Apoptosis ; Polo-like kinase-1