The tumor suppressor function of protein kinase C isoforms (PKC-δ) was evaluated by design and synthesis the 10-23 deoxyribozyme (DRz), thio-modified DRz (DRz-s) and antisense oligonucleotide (asON) of the PKC-δgenomic mRNA to detect the catalytic cleavage activity for tumor cells. Firstly, the cDNA fragment of PKC-δgene was amplified from total cellular RNA of the HepG2.2.15 cells by reverse transcription PCR (RT-PCR). Subsequently, the fragments were cloned to pcDNA3.1(+) plasmids and generated a recombinant plasmids, then sifted the positive recombinant plasmids out to amplify. The expression vector of PKC-δmRNA was obtained in vitro transcription by using T7 RNA polymerase. The results of transfection indicated that when PKC-δmRNA gamyed with deoxyribozyme which activity disappeared, and DRz-s had more intensive specific catalytic cleavage activity than DRz by cells transfecting, but the asON wasn't detected with this activity.
Design and synthesis of triarylacrylonitrile analogues of tamoxifen with improved binding selectivity to protein kinase C
