XP-16, a new xanthono-pyridine derivative, induces apoptosis in human lung carcinoma A549 cells / 中国药理学通报

Aim To investigate the anticancer effect of a new xanthono-pyridine derivative N, N '-( 7-oxo-7H-chromeno[3,2-h] quinoline-5,9-diyl)-bis(2-( pyrroli-din-1-yl)acetamide) (XP-16) on human lung carcino-ma cell line A549 and the potential mechanism. Meth-ods Antiproliferative effect of XP-16 on A549 cells was evaluated by MTT assay, morphological examina-tion and colonial assay. Apoptosis detection was car-ried out using Hoechst 33258 and PI double-dyeing method. Intracellular Ca2+ concentration ( [ Ca2+] i ) and mitochondria membrane potential were detected by fluorospectrophotometer. A549 cells treated with XP-16 were collected for Bad and metallothionein 1 A ( MT-1 A ) transcript analysis by real-time reverse tran-scriptase-polymerase chain reaction ( qRT-PCR) . Re-sults XP-16 inhibited A549 cell proliferation in dose-and time-dependent manner. Typical apoptotic mor- phology such as chromatin aggregation and nuclear fragmentation was observed in A549 cells treated with XP-16 for 24 h, and the apoptosis was showed in a dose-dependent manner. After treated with XP-16, [ Ca2+] i and mitochondria membrane potential of A549 cells were decreased, and relative mRNA level of Bad and MT-1A was up-regulated. Conclusions XP-16 has anticancer effect on A549 cells through apoptosis, which might be associated with decreasing intracellular Ca2+ concentration and mitochondria membrane poten-tial. Up-regulation of MT-1A expression might be the result of decreased [ Ca2+] i .

Interleukin-1-induced iNOS expression in human lung carcinoma A549 cells: Involvement of STAT and MAPK pathways.

For understanding of signaling molecules important in lung cancer growth and progression, IL-1 effect was analyzed on iNOS expression and key signaling molecules in human lung carcinoma A549 cells and established the role of specific signaling molecules by using specific chemical inhibitors. IL-1 exposure (10 ng/ml) induced strong iNOS expression in serum starved A549 cells. Detailed molecular analyses showed that IL-1 increased expression of phosphorylated STAT1 (Tyr701 and Ser727) and STAT3 (Tyr705 and Ser727) both in total cell lysates and nuclear lysates. Further, IL-1b exposure strongly activated MAPKs (ERK1/2, JNK1/2 and p38) and Akt as well as increased nuclear levels of NF-κB and HIF-1 in A549 cells. Use of specific chemical inhibitors for JAK1 kinase (piceatannol), JAK2 kinase (AG-490), MEK1/2 (PD98059) and JNK1/2 (SP600125) revealed that IL-1-induced iNOS expression involved signaling pathways in addition to JAK-STAT and ERK1/2-JNK1/2 activation. Overall, these results suggested that instead of specific pharmacological inhibitors, use of chemopreventive agents with broad spectrum efficacy to inhibit IL-1-induced signaling cascades and iNOS expression would be a better strategy towards lung cancer prevention and/or treatment.

Effective sequence screening of siRNAs targeted protein kinase C ? and their effects on A549 cell line / 基础医学与临床

Objective To investigate the effects of siRNAs targeted protein kinase C ?(PKC?) on the proliferation and apoptosis of A549 cell line.Methods Six PKC? siRNAs were designed and chemical synthesized. Candidate PKC? siRNA was transfected into A549 cells,PKC? mRNA level was determined by semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR). Then,the effects of more effective PKC? siRNAs on A549 were further investigated by using clone formation assay,Hoechst 33258 staining,propidium iodide (PI) staining,Annexin V-FITC and PI dual staining and western blot. Results Six candidate PKC? siRNAs were designed and named as No.1,No.2,No.3,No.4,No.5 and No.6 PKC? siRNA. Compared with controls,PKC? mRNA levels were all significantly downregulated and the cell proliferation was inhibited in A549 cells treated with candidate PKC? siRNAs except No.5 (P

Potential effect and mechanism of IFN? on tumor angiogenesis / 中国临床药理学与治疗学

AIM: To study the effect and potential mechanism of IFN? on tumor angiogenesis. METHODS: Human umbilical cord vascular endothelial cell (HUVEC) was cultured in vitro. The effect of IFN? (10, 100, and 1000 unit?ml -1) on proliferation of HUVEC was detected by MTT assay in vitro. HUVEC pretreated with IFN? (10, 100, and 1000 unit?ml -1) was stained with PI and detected by flow cytometry (FCM). The mRNA expression of apoptosis relative gene Bcl-2 and Bax of HUVEC treated with IFN? (100, and 1 000 unit?ml -1) were detected with semi quantitative reverse transcription- polymerase chain reaction (RT-PCR). IFN? (30, 300, and 3 000 unit?ml -1) on VEGF secretion and expression in human lung carcinoma cell PG in hypoxia were detected by ELISA assay. CONCLUSION: The key attributes of IFN? on tumor angiogenesis possible directly inhibit vascular endothelial cells or indirectly inhibit growth factors secretion and expression in tumor cells.

The expression and its biolgic specification of the Ku80 gene molecular cloning / 中国免疫学杂志

Abstract Objective:To study the expression and its biologic specification of the Ku80 gene molecular cloning.Methods: Constructingrecombinant sense or antisense Ku80 expression vectors (Ku80/pCAGGS) and transferring into LCA .Those Ku80 mRNA and protein were de-teited in the Ku80s-LCA, Ku80as-LCA and by Northern-blotting and or Western-blotting.Results: Those Ku80 mRNA were detected in the Ku80s-LCA and Ku80as-LCA by Northern-blotting. Only the sense Ku80 protein was defected in the Ku80s-LCA , but wasn' t detected in the Ku80as-LCA by Western-blotting.Conclusion:The sense or antisense Ku80 were not only transferred in to LCA ,but also expressed in the mR-NA . Only the sense Ku80 protein was expressed in the Ku80s-LCA , but wasn' t expressed in the Ku80as-LCA . The Ku80 is a candidate tar-get for cancer gene therapy.