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Recombinant mutant human TNF in reversing drug resistance in ovarian cancer cell line SKOV3/DDP and the related mechanism
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Abstract:
To investigate the reversing effect of recombinant mutant human tumor necrosis factor (rmh TNF) on cisplatin(DDP) resistant human ovarian cancer cell line SKOV3/DDP in vitro and the related mechanism. Methods: DDP resistant human ovarian cancer cell line SKOV3/DDP was cultured in vitro. The cytotoxic effect of rmh TNF to SKOV3/DDP cells was examined by MTT assay and the nontoxic dose of rmh TNF was identified. The changes of DDP resistance was observed after cells were treated with nontoxic dose of rmh TNF by MTT assay. The expre ssion of GST π protein was examined by flow cytometry at different periods after rmh TNF intervention; RT PCR was used to analyze the expression of MDR1 gene in SKOV3/DDP cells before and after rmh TNF treatment. Results:（1）rmh TNF at 50 122.34 U/ml showed no evident inhibitory effect on the growth of SKOV3/DDP cells (the cell survival rate higher than 90%); and 100 U/ml was chosen for the reversing experiment ( nontoxic dose).（2）IC 50 values of SKOV3/DDP cells were (2329±0.43), (8.97±0.69) and (6.43±0.79) μg/ml after treatment with DDP for 24, 48 and 72 h, respectively; and the values decreased to （19.50±0.50）,（4.34±0.43） and （2.44±0.02）μg/ml after combined treatment with 100 U/ml rmh TNF, respectively.（3）Expression of GST π protein and MDR1 gene decreased with the prolongation of rmh TNF treatment. Conclusion: rmh TNF has reversal effect on the DDP resistant cell line SKOV3/DDP, and the mechanism may be associated with the down regulation of GST π protein and MDR1 gene expression.
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Project Supported:
Supported by Science and Technology Research Program of Hebei Province(No.072761147)
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