Growth arrest and induction of apoptosis in breast cancer cells by antisense depletion of protein kinase A-RIα subunit: p53-independent mechanism of action

The enhanced expression of the RI subunit of cyclic AMP-dependent protein kinase type 1 (PKA-I) has been correlated with cancer cell growth. We have investigated the effects of sequence-specific inhibition of RI gene expression on the growth of MCF-7 human breast cancer cells. We report that RI antisense treatment results in a reduction in RI expression at both mRNA and protein levels and inhibition of cell growth. The growth inhibition was accompanied by changes in cell morphology, cleavage of poly(ADP-ribose) polymerase (PARP) and appearance of apoptotic nuclei. In addition, bcl-2 protein level was reduced and p53 expression increased in growth arrested cells. Interestingly, RI antisense inhibited cell viability and induced apoptosis in the absence of p53, suggesting that these actions of RI antisense are exerted independent of p53. In contrast, two- and four-base mismatched control oligonucleotides had no effect on either cell growth or morphology. These results demonstrate that the RI antisense, which efficiently depletes the growth stimulatory molecule RI, induces cell differentiation and apoptosis, providing a new approach to combat breast cancer cell growth.