Flow cytometry

Molecular biology

Biology

MTT assay

Apoptosis

Cell growth

Cancer research

Biochemistry

Proliferating cell nuclear antigen

Lupeol

Chemistry

ObjectiveTo investigate the chemosensitivity of pancreatic carcinoma cells to Lupeol.MethodsThe chemosensitivity of cells was determined by the methyl thiazol tetrazolium analysis after exposure to 15 μmol/L Lupeol for 24 h. The results of methyl thiazol tetrazolium (MTT) were further confirmed by morphological analysis. Apoptosis rate was observed by flow cytometry quantitative analysis. Invasion assay was evaluated using transwell chamber assay. The expression of related proteins and mRNAs were detected by Western blotting and reverse transcriptase-polymerase chain reaction (RT-PCR).ResultsAfter pretreatment with Lupeol, half maximal inhibitory concentration of 5-Fu in proliferating cell nuclear antigen (PCNA)-1 cells was reduced from (14.05±2.03) mg/L to (8.58±1.37) mg/L (t=14.355, P=0.005); Meanwhile, pretreatment with Lupeol increased the proliferation inhibition and apoptotic induction of 5-Fu [Apoptosis rate raised from (39.82±4.04)% to (60.43±4.71)%; P=0.000]. The P-gp, MRP and phosphorylated Akt were significantly downregulated after treatment with Lupeol.ConclusionLupeol enhanced the sensitivity of PCNA-1 cells to 5-Fu by inhibiting the expression of P-gp, MRP and the activation of Akt, it might be a potential sensitizer to improve the effects of chemotherapy.Key words: Pancreatic carcinoma; Lupeol; 5-fluorouracil; Chemosensitization

Lupeol enhances the sensitivity of human pancreatic carcinoma cells to 5-fluorouracil