Inhibition of proliferation on Faji cells by the combination of INF-? and curcumin / 中国癌症杂志

Background and purpose:Curcumin is the major effective component of curcuma,curcumin has been noted to be able to selectively inhibit proliferation and induce apoptosis in tumor cells. Interferon-?(IFN-?) is one of the most important cytokines used in clinical treatment nowadays. This study was made to investigate the inhibitory effect of interferon-?(IFN-?)and curcumin on Raji cells (B-NHL) and discuss their mechanism.Methods:The variations of morphology of Raji cells were observed in culture medium after being treated by IFN-?(500,100,2000U/L) combined with various concentrations of curcumin (6.25,12.5,25?mol/L), cells were incubated with the compounds for variable times.The inhibitory ratio was measured by MTT assay, apoptosis was quantitated by flow cytometry(FCM) and the expressions of caspase6, caspase8 and caspase9 in Raji cells were detected by Western blot.Results:both IFN-? and curcumin could significantly induce apoptosis and inhibit the proliferation of Raji cells, synergistic effects were found if cells were treated by the combination of IFN-? and curcumin. The overexpressions of caspase6, caspase8 and caspase9 in Raji cells could be observed in a time-and dose-dependent manner.Conclusions:The combination of IFN-? and curcumin could synergistically inhibit proliferation and induce apoptosis, and its potential mechanism is probably associated with upregulating the expressions of caspase6, caspase8 and caspase9 in B-NHL Raji cell.

zVAD-fmk, unlike BocD-fmk, does not inhibit caspase-6 acting on 14-3-3/Bad pathway in apoptosis of p815 mastocytoma cells

In a preliminary study, we found that benzyloxycarbonyl-Val-Ala-Asp(OMe)-fluoromethylketone (zVAD- fmk), unlike Boc-aspartyl(OMe)-fluoromethylketone (BocD-fmk), at usual dosage could not prevent genistein-induced apoptosis of p815 mastocytoma cells. This study was undertaken to reveal the mechanism underlying the incapability of zVAD-fmk in preventing this type of apoptosis. We observed that 14-3-3 protein level was reduced in genistein-treated cells and that BocD-fmk but not zVAD-fmk prevented the reduction of 14-3-3 protein level and the release of Bad from 14-3-3. We also demonstrated that truncated Bad to Bcl-xL interaction in genistein- treated cells was prevented by BocD-fmk but not by zVAD-fmk treatment. Our data indicate that BocD- fmk, compared to zVAD-fmk, has a certain preference for inhibiting 14-3-3/Bad signalling pathway. We also elucidated that this differential efficacy of BocD-fmk and zVAD-fmk resulted from the different effect in inhibiting caspase-6 and that co-treatment of zVAD-fmk and caspase-6 specific inhibitor substantially prevented genistein-induced apoptosis. Our data shows that caspase-6 plays a role on Bad/14-3-3 pathway in genistein-induced apoptosis of p815 cells, and that the usual dose of zVAD-fmk, in contrast to BocD-fmk, did not prevent caspase-6 acting on 14-3-3/Bad-mediated event.

An immunohistochemical study on the expressions of caspase-6、-7 during the skin incised wound healing in mice / 中国法医学杂志

Objective To investigate the expressions of caspase-6,-7 during skin wound healing and the applicability of the time-dependent expressions of caspase-6,-7 to determination of the wound age.Method The expressions of caspase-6,-7 were studied in skin incised wound in mice by immunohistochemical SABC method,and the non-incised mouse skin was used as control.Results Expressions of caspase-6,-7 were detectable in polymorphonuclear cells (PMNs) in the wound specimens aged 6h. In the wound specimens aged from 12h to 24h after injury, caspase-6,-7 were identified in a large number of infiltrating PMNs and part of mononuclear cells (MNCs). Afterwards, the MNCs and fibroblastic cells (FBCs) accounted for the most part of the caspase-6,-7-positive cells. Morphometrically, the ratios of the number of the caspase-6,-7-stained PMNs, MNCs and FBCs to total number of the cells in the wounds were evaluated and calculated. The ratios of the caspase-6,-7-positive cells were low in the wound specimens aged from 0h to 3h, and maximized in the wound specimens aged 3 day. Thereafter, the ratios decreased and minimized in the specimens aged 14 day.Conclusion The results suggest that caspase-6,-7 were expressed in neutrophils, macrophages and fibroblasts during healing process of skin incised wound in mice. Caspase-6,-7 may be used as a marker for the wound age determination.

Apoptosis in Human Osteosarcoma Cell Line SOSP-9607 Induced by Caspase-6 / 中国肿瘤生物治疗杂志

Objective:To explore caspase-6's effect on the apoptosis of osteosarcoma cell line SOSP 607. Methods: The expression level of caspase-6 in the osteosarcoma cell line SOSP-9067 was examined by RT-PCR method. The adeno-virus adv5 vector with caspase-6 gene was constructed and transferred into the osteosarcoma cell line SOSP-9607 by lipo-fection. The cell survival rate after transfection was assayed by MTT method. The cell morphological changes were observed by microscope and electron microscope, the apoptosis of transferred cells were examined by gel electrophoresis. Results:No expression of caspase-6 was examined in the osteosarcoma cell line SOSP-9607. A high expression of caspase-6 was identified by RT-PCR after the transfection. The cell growth curve declined after transferring caspase-6. Electrophoresis of DNA displayed the apoptosis ladder. Conclusion: Transferring caspase-6 into the osteosarcoma line SOSP-9607 may inhibit the growth of the osteosarcoma cell line SOSP-9607 and this effect might be achieved by inducing apoptosis.

Activation of caspase-6 during resveratrol(Res)-induced apoptosis of nasopharyn- geal carcinoma cells CNE-2Z / 中国免疫学杂志

Objective:To study whether caspase-6 was activated during resveratrol(Res)- induced apoptosis of nasopharyngeal carcinoma cells CNE-2Z. Methods:The cleavage of pro-caspase-6 was analyzed by Western-blot. TTie changes of caspase-6 mKNA was detected by semi-quantitative RT-PCR. The changes of caspase-6 activity was determined by colorimetric assay. Results: After CNE-2Z cells were treated with 0 (control), 25, 50, 100, 200 pmol/L Res for 24 h, respectively, the expression of pro-caspase-6 was decreased with concentration increasing. Caspase-6 active fragment P20(20 kD) appeared at 100 fjtmol/L and was increased at 200 (junol/L.The expression of caspase-6 mR-NA was increased in a concentration-dependent manner ( P