[Apoptosis of human gastric cancer cells induced by bcl-2 antisense oligodeoxynucleotides].
OBJECTIVE
To study the regulation of bcl-2 gene expression and induction of apoptosis by bcl-2 antisense oligodeoxynucleotides (AS-ODN) on human gastric cancer cell line BGC-823 in vitro.


METHODS
Two bcl-2 AS-ODNs were synthesized, one covering the initiation sequence of translation of bcl-2 mRNA (AS-ODN1) and the other covering the protein coding region (AS-ODN2). BGC-823 cells in logarithmic phase of growth were cultured in the presence of free or liposome (DOTAP)-encapsulated AS-ODN. Cell growth was assessed by MTT method. The expression of bcl-2 at mRNA and protein levels was examined by RT-PCR and flow cytometry, respectively. Electron microscopy and flow cytometry were used to demonstrate apoptotic changes in AS-ODN-treated cells.


RESULTS
Both AS-ODNs inhibited proliferation of BGC-823 cells. The inhibitory activity of AS-ODN2 was stronger than that of AS-ODN1. AS-ODNs encapsulated in liposome led to more marked inhibition of cell growth than free AS-ODNs. Both AS-ODNs reduced bcl-2 expression of BGC-823 cells at mRNA and protein levels. Apoptosis of BGC-823 cells were demonstrated by the appearance of apoptotic bodies, chromatin condensation and pre-G1 peak on flow cytometric analysis.


CONCLUSIONS
Antisense oligodeoxynucleotide of bcl-2 decreases bcl-2 gene expression and induces apoptosis of human gastric cancer cells in vitro.
Lu
P
P
First Affiliated Hospital, China Medical University, Shenyang 110001.
Chen
J
J
D017209:Apoptosis / Q000187:drug effects*; D019254:Genes, bcl-2 / Q000502:physiology*; D006801:Humans; D020319:Oligodeoxyribonucleotides, Antisense / Q000494:pharmacology*; D012333:RNA, Messenger / Q000032:analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013274:Stomach Neoplasms / Q000188:drug therapy / Q000235:genetics / Q000473:pathology*; D014407:Tumor Cells, Cultured
D020319:Oligodeoxyribonucleotides, Antisense; D012333:RNA, Messenger