JS­K induces G2/M phase cell cycle arrest and apoptosis in A549 and H460 cells via the p53/p21WAF1/CIP1 and p27KIP1 pathways.

Lung cancer is one of the most common malignancies worldwide, with high mortality and morbidity rates. O2­â€‹(2,4­â€‹dinitrophenyl)­1­â€‹[(4­ethoxycarbonyl)piperazin­1­yl]diazen­1­ium­1,2­diolate (JS­K) is a potent anticancer agent that acts against a subset of human non­small cell lung cancer (NSCLC) cell lines; however, the underlying mechanisms of JS­K in NSCLC remain unclear. The present study aimed to evaluate the anticancer effect of JS­K and investigate its underlying mechanisms in A549 and H460 cells. In the present study, A549 and H460 cells were treated with JS­K, and then evaluated by cell viability assay, flow cytometry and western blot analysis. JS­K markedly induced cell cycle arrest at the G2/M phase in a concentration and time­dependent manner in both cell lines. This was associated with increased expression levels of p53, and the cell cycle inhibitors p21WAF1/CIP1 and p27KIP1, which, in turn, inhibited the expression of Cdc2, cyclin B1 and cyclin­dependent kinase 2. In addition, JS­K­induced inhibition of proliferation was revealed to be partially modulated by the upregulation of p53 and p21WAF1, the ratio of Bax/Bcl­2, and the activation of both the intrinsic and extrinsic apoptotic pathways in A549 and H460 cells. These results demonstrated that JS­K could trigger cell cycle arrest at the G2/M phase and apoptosis in A549 and H460 cells, which was likely mediated via the p53/p21WAF1/CIP1 and p27KIP1 pathways. Overall, the results indicated that JS­K may be used as an anticancer agent for the treatment of NSCLC.

JIB­04 induces cell apoptosis via activation of the p53/Bcl­2/caspase pathway in MHCC97H and HepG2 cells.

JIB­04 is a structurally unique small molecule, known to exhibit anticancer activity and to inhibit the growth of human lung cancer and prostate cancer cell lines. However, the anticancer effect of JIB­04 against human hepatic carcinoma, and its underlying mechanisms, are still unclear. In the present study, MHCC97H and HepG2 cells were employed to investigate the anticancer effects of JIB­04 on cell viability and apoptosis. Annexin V/PI staining, a CCK­8 assay and western blot analysis demonstrated that JIB­04 induced apoptosis in MHCC97H and HepG2 cells, which was evidenced by the expression of proapoptotic and apoptotic proteins including p53, Bak, Bax, caspase­3 and caspase­9. Subsequently, the expression trends of Bcl­2 and p53 were reversed after co­treatment with pifithrin­α (PFT­α, a p53 inhibitor). The results revealed that JIB­04 suppressed the cell viability of MHCC97H and HepG2 cells in a concentration­dependent manner. Meanwhile, it was also demonstrated that JIB­04 effectively triggered MHCC97H and HepG2 cell apoptosis by downregulating Bcl­2/Bax expression, and upregulating proapoptotic and apoptotic protein expression via the p53/Bcl2/caspase signaling pathway. JIB­04 had effects on the inhibition of cell viability and the induction of apoptosis in MHCC97H and HepG2 cells. The underlying mechanism of action of JIB­04 was associated with the p53/Bcl­2/caspase signaling pathway. Our findings provide a foundation for understanding the anticancer effect of JIB­04 on MHCC97H and HepG2 cells, and suggested that JIB­04 may be a promising therapeutic agent in human liver cancer.

In vivo microdialysis with ultra performance liquid chromatography-mass spectrometry for analysis of tetramethylpyrazine and its interaction with borneol in rat brain and blood.

Tetramethylpyrazine (TMP) has been widely used in the treatment of ischemic cerebrovascular disease. However, the mechanism of TMP and how to increase its bioavailability need to be further explored. In our study, an in vivo microdialysis sampling technique coupled with ultra-performance liquid chromatography-mass spectrometry method was developed to investigate the pharmacokinetic properties of TMP and its interaction with different doses of borneol (BO) in rats. Linearity of TMP in brain and blood dialysates exhibited good linear relationships over the concentration range of 0.991-555.14 ng/mL. The specificity, linearity, accuracy, precision, matrix effect and stability were within acceptable ranges. The results demonstrated that BO had a marked impact on the pharmacokinetic properties of TMP. After co-administration, the areas under the concentration-time curve (AUC) of TMP in brain and blood were significantly increased. Meanwhile, the peak concentration of TMP in brain was also enhanced. The AUCBrain /AUCBlood of TMP, increased from 44% to 56 and 60.8% after co-administration with BO (15 and 30 mg/kg). The pharmacodynamic results showed that TMP co-administration with BO enhanced the cerebral blood flow during the period of ischemia and reduced the infarct volume. Overall, it might be an effective way to treat stroke to use TMP co-administered with BO.

Aberrant histone modification in peripheral blood B cells from patients with systemic sclerosis.

OBJECTIVES: To investigate alterations in histone modifications in B cells and their role in the pathogenesis of systemic sclerosis (SSc). METHODS: Global histone H3/H4 acetylation and H3K4/H3K9 methylation in B cells of SSc were tested by EpiQuik™ assay kits. Related histone modifier enzymes were measured by RT-PCR and Western blot. RESULTS: Global histone H4 hyperacetylation and global histone H3K9 hypomethylation were observed in SSc B cells compared with controls. Expression of JHDM2A was significantly increased but HDAC2, HDAC7, and, SUV39H2 were significantly down-regulated in SSc B cells relative to controls. Global histone H4 acetylation and the expression of HDAC2 were negatively correlated. Global histone H3K9 methylation and the expression of SUV39H2 protein were positively correlated. Global H4 acetylation was positively correlated with disease activity and expression of HDAC2 protein was negatively correlated with skin thickness. CONCLUSIONS: Histone modifications were altered in B cells in SSc correlating with skin thickness and disease activity.