Antitumor effect of PUMA overexpression on pancreatic cancer ASPC-1 cells

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Abstract

Objective

To investigated the antitumor effects of PUMA gene transfection on pancreatic cancer Aspc-1 cells.

Methods

Plasmid pGFP-PUMA-C1 and pGFP-C1 was introduced into the pancreatic cancer ASPC-1 cells by LipofectinamineTm 2000 transfection. 24 h and 48 h after transfection, these cells were collected, PUMA protein and PUMA mRNA expression in ASPC-1 cells were detected by Western blot and semiquantitative reverse transcription polymerase chain reaction (RT-PCR) methods, respectively. Cell apoptosis was examined by flow cytometry and terminal deoxynucleotidyl transferase(TdT)-mediated dUTP nick end labeling (TUNEL). Growth inhibition of Aspc-1 cells was determined by the colorimetric MTT cell Viability/proliferation assay.

Results

Transfection of pGFP-PUMA-C1 into Aspc-1 cells resulted in the upregulation of the corresponding mRNA and PUMA protein, which was associated with a reduced number of viable cells and increased number of apoptosis cells, but the mRNA and PUMA protein and the corresponding viable cells and apoptosis cells had no significant differences in Aspc-1 cells/pGFP-C1 compared to control cells.

Conclusion

Re-expression of PUMA gene, which is lost in human pancreatic cancer cells, can induce apoptosis, resulting in inhibition of tumor growth.

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