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AIM: To investigate the expression of genes involved in the gemcitabine-induced cytotoxicity in human pancreatic cancer cells. METHODS: A human pancreatic cancer cell line, PANC-1, was cultured. 1×104 PANC-1 cells were plated in 96-well microtiter plates. After being incubated for 24 h, gemcitabine was added to the medium at concentrations ranging 2.5 -1 000 mg/L. The AlamarBlue dye method was used for cell growth analysis. DNA fragmentation was quantitatively assayed using a DNA fragmentation enzyme-linked immunosorbent assay (ELISA) kit. PAP and TP53INP1 mRNA expression was determined using the reverse transcription-polymerase chain reaction with semi-quantitative analysis. The expression of GSK-3βand phospho-GSK-3βproteins was examined with Western blot analysis. RESULTS: The IC50 for the drug after a 48-h exposure to gemcitabine was 16 mg/L. The growth of PANC-1 cells was inhibited by gemcitabine in a concentrationdependent manner (P< 0.0001) and the cell growth was also inhibited throughout the time course (P< 0.0001). The DNA fragmentation rate in the gemcitabine-treated group at 48 h was 44.7 %, whereas it was 25.3 % in the untreated group. The PAP mRNA expression was decreased after being treated with gemcitabine, whereas the TP53INP1 mRNA was increased by the gemcitabine treatment. Western blot analysis showed that phosphoGSK-3βser9 was induced by the gemcitabine treatment. CONCLUSION: Gemcitabine suppresses PANC-1 cell proliferation and induces apoptosis. Apoptosis is considered to be associated with the inhibition of PAP and GSK-3β, and the activation of TP53INP1 and pospho- GSK-3βser9.
AIM: To investigate the expression of genes involved in the gemcitabine-induced cytotoxicity in human pancreatic cancer cells. METHODS: A human pancreatic cancer cell line, PANC-1, was cultured. 1 × 104 PANC- microtiter plates. After being incubated for 24 h, gemcitabine was added to the medium at concentrations ranging from 2.5 to 1000 mg / L. The Alamar Blue dye method was used for cell growth analysis. DNA fragmentation was quantitatively assayed using a DNA fragmentation enzyme-linked The expression of GSK-3βand phospho-GSK-3βproteins was examined with Western blot analysis. RESULTS: The IC50 was used in immunosorbent assay (ELISA) kit. PAP and TP53INP1 mRNA expression was determined using the reverse transcription-polymerase chain reaction with semi-quantitative analysis for the drug after a 48-h exposure to gemcitabine was 16 mg / L. The growth of PANC-1 cells was inhibited by gemcitabine in a concentration dependent manner (P <0.0001) and the cell growth was also inhibit The DNA fragmentation rate in the gemcitabine-treated group at 48 h was 44.7%, but it was 25.3% in the untreated group. The PAP mRNA expression was decreased after being treated with gemcitabine, but the TP53INP1 mRNA was increased by the gemcitabine treatment. Western blot analysis showed that phosphoGSK-3βser9 was induced by the gemcitabine treatment. CONCLUSION: Gemcitabine suppresses PANC-1 cell proliferation and induces apoptosis. Apoptosis was considered to be associated with the inhibition of PAP and GSK-3β, and the activation of TP53INP1 and pospho-GSK-3βser9.