Phosphorylated extracellular signal-regulated kinase up-regulated p53 expression in shikonin-induced HeLa cell apoptosis / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>The role of extracellular signal-regulated kinase 1/2 (ERK1/2) in shikonin-induced HeLa cells apoptosis remains vague. This study was to investigate the activation of caspase pathways and the role of ERK1/2 in human cervical cancer cells, HeLa, by shikonin.</p><p><b>METHODS</b>The inhibitory effect of shikonin on the growth of HeLa cells was measured by MTT assay. Fluorescent microscopic analysis of apoptotic cells stained with 4',6'-oliiamiclino-2-phenylindole C (DAPI) and Hoechst 33258 was carried out. Caspase-3 and -8 activities were detected using caspase-3 substrate and caspase-8 substrate as substrates, respectively. The protein levels of ERK, p53 and p-ERK were determined by Western blot analysis.</p><p><b>RESULTS</b>Shikonin inhibited cell growth in a time- and dose-dependent manner. Caspase-3 and caspase-8 were activated in the apoptotic process and caspase inhibitors effectively reversed shikonin-induced apoptosis. Phosphorylation of ERK resulted in up-regulation of p53 expression, which was blocked by mitogen-activated protein kinase (MEK), inhibitor PD 98059.</p><p><b>CONCLUSION</b>Shikonin induces HeLa cell apoptosis through the ERK, p53 and caspase pathways.</p>

Pseudolaric acid B induced apoptosis and mitotic arrest circumventing Fas receptor pathway in MCF-7 cells / 中国药理学通报

Aim To study the mechanisms of Pseudolaric acid B(PAB)-induced MCF-7 cell apoptosis and mitotic arrest.Methods MTT assay was performed to assess the cell growth inhibition,contrast phase microscope was used to observe cellular morphologic alteration,and the change of DNA was detected by fluorescent microscopy.The distribution of cell cycle was determined by flow cytometric analysis of propidium iodide staining,and the protein expression was examined by Western blot analysis.Results PAB inhibited MCF-7 cell growth in a dose-and time-dependent manner.4 ?mol?L-1 PAB induced DNA condensation at 24 h.PAB cleaved PARP in a time-dependent manner.At 36 h,PAB up-regulated the expression of cdc 2 and nuclear cyclin B1.Fas antagonistic antibody UB2 had no effect on apoptosis,but agonistic antibody CH11 enhanced the apoptosis induced by PAB.UB2 exerted no effect on cell cycle arrest,and CH11 had the same action as UB2 except for reducing the mitotic arrest through enhancing apoptotic subdiploid peak.Conclusion PAB inhibited MCF-7 cell growth through mitotic arrest and apoptosis.Apoptosis and mitotic arrest were independent of Fas pathway.

The Clinical Usage and Dosage Form of Dai-kenchu-to Extract / 日本東洋医学雑誌

All prescriptions containing Dai-kenchu-to (DKT, Da-Jian-Zhong-Tang), one of the most commonly prescribed Kampo medicines (Sino-Japanese traditional herbal medicines) in Japan, that had been issued during a six-month period from September 1999 to February 2000 at Kitasato University Hospital, were analyzed. The DKT used at this hospital was in the form of ethical extract granules (Tsumura, Tokyo), and it was mainly prescribed to patients who were treated in the Departments of Gynecology and Surgery and who had undergone surgery. In order to clarify problems in the administration of DKT granules to post-operative patients, a questionnaire was distributed to nurses working in the wards of eight large hospitals (over 470 beds each). More than 80% of the nurses reported that they had observed problems with the taste and dosage of DKT administered to their patients. Hard granules do not readily dissolve in water, and the nurses observed difficulty in deglutition upon oral administration of DKT, and tube obstruction in patients who were administrated DKT through a gastric tube. These difficulties in the administration of DKT are thought to increase the workload of the care staff, and the development of a new route of administration of DKT, for example, a decocted solution packed into a stick, is expected.

Regulation of ERK MAPK in evodiamine-induced A375-S2 cell death / 中国病理生理杂志

AIM: To compare the cytotoxic effect of evodiamine with chemotherapy drugs on A375-S2 cells, and to examine the relationship between the effects of PKC and ERK on evodiamine-induced cell death. METHODS: MTT assay and Western blot analysis were applied. RESULTS: Compared to actinomycin D, cisplatin and 5-FU, evodiamine showed less cytotoxic effects on A375-S2 cells, but it induced more significant inhibition of proliferation in A375-S2 cells incubated with evodiamine for 24 h, followed by continuous culture in drug-free medium. The activation of PKC induced by 10 ?g?L -1 PMA partially blocked evodiamine-induced cell death, which was reversed by PKC and ERK inhibitors. Moreover, evodiamine down-regulated the expressions of ERK and phosphorylated ERK. CONCLUSION: Evodiamine has a strong inhibitory influence on proliferation of A375-S2 cells, even after removal of evodiamine. Evodiamine blocks the protective role of ERK to A375-S2 cells through the downregulation of ERK and phosphorylated ERK expression. [

Human interleukin-1? induces A375-S2 human melanoma apoptosis through caspase pathway / 中国病理生理杂志

AIM: To study the molecular biological mechanism and signal transduction pathway of interleukin-1? (IL-1?)-induced apoptosis in A375-S2 melanoma cells. METHODS: Photomicrocropy showed typical apoptotic changes. The cytotoxic effect of IL-1? in vitro and influences of caspases in this effect were measured by MTT assay. The cytotoxicity of cells was assessed by LDH-based assay. Degradation of DNA was detected by agarose gel electrophoresis. RESULTS: The inhibitory effect of IL-1? on A375-S2 cell growth was in a dose and time-dependent manner, and cell death rate reached more than 90% at 72 h after treatment with 10~(-9)mol/L IL-1?. The inhibitors of caspase-family, -1, -3, -8, -9, and -10, partially blocked cell death at early stage. LDH assay showed that major IL-1?-induced cell death was apoptosis, and in a dose and time-dependent manner. Typical apoptotic DNA ladder was observed in agarose gel electrophoresis. CONCLUSION: IL-1? induced apoptosis in melanoma A375-S2 cells by activating caspase pathway. [

Magnolol induces apoptosis via alteration of Bax/Bcl-X_L ratio and activation of caspases / 中国药理学通报

Aim To study the mechanism of magnolol-induced HeLa cell death.Methods Cell viability was measured by MTT method.Morphological changes were observed by phase contrast microscopy and Hoechst 33258 staining. DNA fragmentation was assayed by agarose gel electrophoresis. Protein level was detected by Western blot analysis.Results Magnolol induced HeLa cell apoptosis and had a weaker cytotoxic effect on HEL 299 cell. The apoptosis of HeLa cells was partially reversed by caspase-3,-8,-9,-10 and caspase family inhibitors. Magnolol has a synergistic apoptotic effect with Fas agonistic antibody CH-11. Treatment of HeLa cells with magnolol for 12 h increased the protein expression ratio of Bax/Bcl-X_L; procaspase-3, ICAD and PARP were cleaved to smaller molecules. p53 and phosphorylation of p53 (ser-15) increased response to magnolol treatment.Conclusion Magnolol induced HeLa cell death via alteration of Bax/Bcl-X_L ratio,activation of caspases and accumulation of p53.

Shikonin induce HeLa cell apoptosis via a caspase-dependent mechanism / 中国药理学通报

AIM To study the mechanisms of shikonin-induced HeLa cell apoptosis. METHODS Cytotoxicity assay by MTT, DNA fragmentation assay, LDH activity-based cytotoxicity assay and Western blot analysis were carried out. RESULTS Shikonin inhibited HeLa cell growth in a dose- and time-dependent manner. Apoptotic cell death was observed within 24 h after 40 ?mol?L -1 shikonin treatment. Shikonin induced HeLa cell apoptosis involved in the activation of caspase-3 and caspase-8, which is similar to the Fas or TNF-? signaling transduction pathways. CONCLUSION Shikonin induced caspase-dependent apoptosis in HeLa cells.

Oridonin induced HeLa cell apoptosis through capase pathway / 中国药理学通报

AIM To study the mechanisms of oridonin-induced HeLa cell apoptosis. METHODS Morphological observation, agarose gel electrophoresis, LDH release and caspase inhibitors were used. RESULTS Oridonin had significant apoptotic effect on HeLa cells in a dose- and time-dependent manner. The marked morphological changes including condensed chromatin, nuclear fragmentation and apoptotic bodies and DNA ladder in agarose gel was observed. The activity of caspase-3 was increased about 2 times of control value after treatment with oridonin. CONCLUSION Oridonin induces HeLa cell apoptosis through caspase pathway.

Oridonin enhances phagocytosis of apoptotic U937 cells by macrophage-like cells / 中国病理生理杂志

AIM: To study the effect of oridonin on the phagocytosis of apoptotic U937 cells by macrophage-like cells. METHODS: DNA agarose gel electrophoresis, Giemsa staining, Hoechst 33258 staining and photomicroscopical observation were used. RESULTS: UV irradiation (2.4 J/cm~2, 4 min) induced U937 cell apoptosis. Marked DNA fragmentation in agarose gel electrophoresis was observed. Oridonin augmented phagocytosis of apoptotic U937 cells by U937 cell-derived macrophages in a time- and dose-dependent manner. However, less effect on synthetic fluoresbrite micropheres was observed. The oridonin-augmented phagocytosis was attenuated by anti-human TNF? or anti-human IL-1? antiserum. In addition, the similar effect of phagocytosis was observed in oridonin-treated human monocyte-derived macrophages at 4 day maturation. CONCLUSION: Oridonin enhances phagocytosis of apoptotic U937 cells by macrophage-like cells. The releases of TNF? and IL-1? are involved in this mechanism.

Norcantharidin induces HeLa cells apoptosis through caspases pathway / 中国病理生理杂志

AIM: To examine the apoptotic pathway of norcantharidin (NCTD)-induced HeLa cells death. METHODS: MTT, photomicroscopical observation, DNA agarose gel electrophoresis, LDH release and Western blot analysis were used. RESULTS: NCTD induced HeLa cells apoptosis and the apoptosis was partially reversed by the inhibitors of caspase-family (-3, -8, -10). The activities of caspase-3, -8 and -9 were significantly increased after treated with NCTD. The expression of the inhibitor of caspase-3 activated DNase (ICAD) was decreased in a time dependent manner. CONCLUSION: NCTD induces HeLa cells apoptosis through activating caspase pathways.