Perilla leaf, Perilla frutescens, induces apoptosis and G1 phase arrest in human leukemia HL-60 cells through the combinations of death receptor-mediated, mitochondrial, and endoplasmic reticulum stress-induced pathways.

Since it has been reported that Perilla leaves (Perilla frutescens) have antimutagenic, antioxidant, and anti-inflammatory properties, we hypothesized that Perilla leaves may have a potential anticancer activity. Therefore, we examined the possibility that cancer cell growth is reduced by treatment with a Perilla leaf ethanol extract (PLE) using human leukemia HL-60 cells and then investigated the mechanism of the growth inhibition. We found that PLE treatment suppressed cell viability in a dose-dependent manner. Flow cytometric analysis revealed that PLE treatment caused the appearance of a sub-G1 DNA peak and induced cell cycle arrest at the G1 phase. We detected DNA ladders in PLE-treated cells by agarose gel electrophoresis, and the cleavage of pro-caspase-3 and poly(ADP-ribose) polymerase with remarkable activation of caspase-8, -9, and -3. Western blot analysis revealed dose-dependent increases in Bax and cytochrome c in cytosol fractions and decreased Bid and pro-caspase-8 and -3 in PLE-treated cells. In addition, glucose-regulated protein 78, phosphorylated eukaryotic translation initiation factor 2 subunit alpha, phosphorylated c-jun N-terminal kinase, and p21 levels were increased by PLE treatment in a dose-dependent manner, whereas the p27 level was not changed. We concluded that PLE induced apoptosis through the combinations of mitochondrial, death receptor-mediated, and endoplasmic reticulum pathways and suppressed the cell proliferation via p21-mediated G1 phase arrest in HL-60 cells.

Application of the multiplex PCR method for discrimination of Artemisia iwayomogi from other Artemisia herbs.

Some plants classified in the genus Artemisia are used for medicinal purposes. In particular, A. iwayomogi, which is referred to as 'Haninjin,' is used as an important medicinal material in traditional Korean medicine. However, A. capillaris, and both A. argyi and A. princeps, referred to as 'Injinho' and 'Aeyup,' respectively, are used for purposes other than those for which 'Haninjin' is utilized. However, it is occasionally difficult to differentiate 'Haninjin' from 'Injinho' and/or 'Aeyup' on the basis of their morphological features, particularly when in the dried and/or sliced form. Therefore, the development of a reliable method by which to discriminate 'Haninjin' from other Artemisia herbs, especially 'Injinho' and 'Aeyup,' is clearly necessary. We recently determined that the RAPD (random amplified polymorphic DNA) technique can be used to discriminate efficiently between some Artemisia herbs. In particular, when applied to RAPD, the non-specific UBC primer 391 (5'-GCG AAC CTC G-3') was demonstrated to amplify PCR products specific to A. iwayomogi. Based on the nucleotide sequences of the PCR product, we designed a 2F1 (5'-ACC TCG GAC CTA AAT ACA-3')/ 2F3 (5'-TTA TGA TTC ATG TTC AAT TC-3') primer set to amplify a SCAR (sequence-characterized amplified region) marker of A. iwayomogi. Employing this primer set, along with two other primer sets amplifying SCAR markers of 'Aeyup' (A. argyi and A. princeps) and both 'Injinho' (A. capillaris) and A. japonica, which are classified into the same subgroup in a phenogram constructed from RAPD analysis, we developed a multiplex PCR method by which A. iwayomogi could be discriminated with certainty from other Artemisia herbs. Via this method, we determined not only whether the tested Artemisia herb was A. iwayomogi, but also which Artemisia herbs were tested concurrently with A. iwayomogi.