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Inhibition of Bcl-2 Protein Expression and Induction of Apoptosis in Renal Cell Carcinoma Cells by Antisense Oligodeoxynucleotide Targeting the bcl-2 Gene
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Abstract:
Objective:To evaluate the effects of antisense oligodeoxynucleotides (ODNs) (AS1 complementary to the translation initiation region and AS2 complementary to the coding region) targeted to bcl-2 oncogene on Bcl-2 protein expression and apoptosis of human renal cell carcinoma (RCC) cells.Methods:Expression of bcl-2 mRNA in RCC cell lines was analyzed by reverse transcriptase-polymerase chain reaction (RT-PCR). The ODNs were transfected with Lipofectin into RCC cell lines. The expression of Bcl-2 protein in ACHN tumor cells was examined by Western blot analysis, and the apoptosis of those cells was determined by flow cytometric analysis.Results: Expression of bcl-2 mRNA was detected in all five RCC lines. Transfected bcl-2 antisense ODNs, but not sense ODNs, inhibited Bcl-2 protein expression in ACHN cells. The AS2 antisense ODN showed a superior effect compared with AS1 ODN. The apoptosis of ACHN cell could been induced by bcl-2 antisese ODNs , and percentage of apoptotic cells was noted 32.1% and 43.2% treated with AS1 and AS2, respectively. Conclusions:Treatment of human RCC cells with antisense ODNs targeting bcl-2 gene inhibits expression of Bcl-2 protein and induce apoptosis.
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R730.59 R735.7
