TNF相关凋亡诱导配体联合阿糖胞苷诱导HL-60细胞凋亡及其机制的研究

本研究探讨人重组可溶性肿瘤坏死因子相关凋亡诱导配体（hrsTRAIL）单用或联合阿糖胞苷（Ara—C）诱导HL-60细胞凋亡作用及其机制。在体外以人类急性白血病细胞株HL-60为研究对象，分5组进行细胞培养：对照组、Ara—C组、rsTRAIL组、同时给药组（Ara—C＋rsTRAIL）、先后给药组（Ara—C→rsTRAIL）。采用MTT比色法测定细胞生长抑制率；应用Annexin V／PI染色和流式细胞术检测细胞凋亡率；使用流式细胞术检测不同浓度的Ara—C对细胞表面DR5表达水平的影响及rsTRAIL单药组、先后给药组细胞表面DR5表达水平和细胞内caspase-8活性。结果表明：rsTRAIL能抑制HL-60细胞生长并诱导HL-60细胞凋亡。先后给药组诱导HL-60细胞凋亡率大于同时给药组。先后给药组细胞表面DR5表达水平和细胞内caspase-8的活性高于rsTRAIL组。5mg／L、10mg／LAra—C作用于HL-60细胞24小时，其细胞表面DR5表达水平升高，大于对照组。结论：rsTRAIL抑制HL-60细胞生长．诱导HL-60细胞凋亡。Ara—C上调HL-60细胞表面DR5表达水平。增强rsTRAIL诱导HL-60细胞凋亡的作用。Ara—C和rsTRAIL联合用药时，在先加Ara—C、再加rsTRAIL组诱导HL-60细胞凋亡的效果比同时给药组效果好，其机制可能与Ara—C上调细胞表面DIL5表达水平和（或）增强细胞内caspase-8的活性有关。     

化疗药物对TRAIL在急性白血病原代细胞表达的影响

为了探讨TRAIL在急性白血病原代细胞表达及化疗药物对TRAIL在急性白血病细胞表达的影响,采集16例初诊的急性白血病患者化疗前、化疗1天后、3天后和5天后的外周血,分离单个核细胞并用流式细胞术检测TRAIL的表达;同时采集12例初诊的急性白血病患者化疗前骨髓,分离单个核细胞进行培养,分别加VP-16和/或干扰素,培养一定时间后检测TRAIL的表达水平.结果表明：与化疗前相比,初治急性白血病患者外周血单个核细胞TRAIL的表达水平在化疗1天后即明显提高（P＜0.05）;在体外培养实验中,VP-16上调TRAIL在急性白血病骨髓单个核细胞的表达（P＜0.05）,VP-16联合干扰素与单独应用VP-16相比,TRAIL的表达水平无差异（P＞0.05）.结论：化疗药物治疗白血病的机制之一可能是通过诱导TRAIL的表达而促进白血病细胞的凋亡.     

TRAIL受体DR4、DR5、DcR1在结肠癌中的表达及意义

目的 探讨肿瘤坏死因子相关凋亡导配体(TRAIL)受体DR4、DR5、DcR1在结肠癌中的表达及临床意义,为TRAIL的抗肿瘤作用提供实验依据.方法 采用免疫组化SP法检测35例结肠癌和14例正常结肠黏膜组织中DR4、DR5、DcR1表达水平.结果 35例结肠癌组织中DR5的阳性表达率为27/35(77.14%),DR...     

Chrysin promotes tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL) induced apoptosis in human cancer cell lines

Chrysin exists widely in plants, honey and propolis. The anti-cancer property of chrysin has been demonstrated though the molecular mechanism is not clear. In this study, we found that pre-treatment with chrysin could promote the cell death induced by TRAIL according to the morphological changes and appearance of sub-G1 peak in four human cancer cell lines. In HCT-116 cells, the results of flow cytometry analysis showed that the percentage of sub-G1 reached (38.89 ± 3.78) % when pre-treatment of chrysin was used at 40 μM, but that was only (2.53 ± 0.10) % in the untreated group and (13.22 ± 0.20) % in TRAIL alone group. The differences between the combination and the untreated or TRAIL alone group were all significant (P < 0.05) and dose-dependent effect was obvious. Similar results were obtained in CNE1 cells. In the search of molecular mechanisms, we found that pre-treatment with chrysin could increase TRAIL-induced degradation of caspase 3, caspase 8, PARP proteins. Z-VAD-fmk, which is a pan-caspase inhibitor, could inhibit the apoptosis enhanced by the combination of chrysin and TRAIL. All data indicate that chrysin can enhance the apoptosis induced by TRAIL, and the apoptosis is caspase-dependent and related to the activation of caspase 8.     

痛风性关节炎的生物标志物

目的筛选大鼠痛风性关节炎(GA)生物标志物。方法采用改良的微晶尿酸钠和次黄嘌呤法制备大鼠急性GA模型,检测血尿酸、尿尿酸、24 h总尿量及滑膜组织病理形态学改变,采用RayBioBiotin Label-based Rat Antibody ArrayⅠ筛选差异表达蛋白。结果急性GA模型建立成功,发现GA中14个差异表达蛋白,其中9个细胞因子首见于GA,差异最明显2个蛋白为TRAIL与Neuropilin-2。结论 TRAIL与Neuropilin-2为GA生物标志物,为临床诊断与防治提供新靶点。     

Effect of Smac on TRAIL-induced apoptosis of prostate cancer cell line PC-3 and the molecular mechanism

The effect of Smac gene on the TRAIL-induced apoptosis of the prostate cancer cell line PC-3 and the molecular mechanism were investigated. The Smac gene was transfected into PC-3 cells under the induction of liposome. The intrinsic Smac gene expression was detected by Western blotting. After treatment with TRAIL as an apoptosis inducer, in vitro cell growth activity was as-sayed by MTT colorimetry. The apoptosis rate of PC-3 cells was determined by annexin Ⅴ-FITC and propidium iodide staining flow cytometry. The expression of cellular XIAP and caspase-3 genes was examined by Western blotting. Smac-transfected cells (PC-3/Smac group) had significantly in-creased Smac protein level as compared with PC-3 controls (P<0.01). After induction with 100-200 ng/mL TRAIL for 12-36 h, cellular proliferation rate in PC-3/Smac group was significantly lower than in PC-3 controls (P<0.05). After induction with 100 ng/mL TRAIL for 24 h, the apoptosis rate in PC-3/Smac group was significantly enhanced as compared with that of PC-3 controls (P<0.05). Ac-cordingly, the XIAP expression level was down-regulated significantly (P<0.05) and caspase-3 sub-unit P20 was up-regulated significantly (P<0.05). It is suggested that the over-expression of cellular Smac can inhibit inhibitor of apoptosis proteins (IAPs), enhance caspases activity and the apoptosis rate of PC-3 cells induced by TRAIL, which may provide a useful experimental basis for prostate cancer therapy.     

Apigenin Sensitizes Huh-7 Human Hepatocellular Carcinoma Cells to TRAIL-induced Apoptosis

TNF-related apoptosis-inducing ligand (TRAIL) is a promising agent for management of cancer because of its selective cytotoxicity to cancer cells. However, some cancer cells have resistance to TRAIL. Accordingly, novel treatment strategies are required to overcome TRAIL resistance. Here, we examined the synergistic apoptotic effect of apigenin in combination with TRAIL in Huh-7 cells. We found that combined treatment of TRAIL and apigenin markedly inhibited Huh-7 cell growth compared to either agent alone by inducing apoptosis. Combined treatment with apigenin and TRAIL induced chromatin condensation and the cleavage of poly (ADP-ribose) polymerase (PARP). In addition, enhanced apoptosis by TRAIL/apigenin combination was quantified by annexin V/PI flow cytometry analysis. Western blot analysis suggested that apigenin sensitizes cells to TRAIL-induced apoptosis by activating both intrinsic and extrinsic apoptotic pathway-related caspases. The augmented apoptotic effect by TRAIL/apigenin combination was accompanied by triggering mitochondria-dependent signaling pathway, as indicated by Bax/Bcl-2 ratio up-regulation. Our results demonstrate that combination of TRAIL and apigenin facilitates apoptosis in Huh-7 cells.     

高血糖对小鼠卵母细胞Ca2+振荡和卵泡颗粒细胞肿瘤坏死因子相关凋亡诱导配体表达的影响

目的 探讨高血糖对小鼠卵母细胞Ca2+振荡和卵泡颗粒细胞肿瘤坏死因子相关凋亡诱导配体(TRAIL)表达的影响.方法 出生后20d ICR雌鼠,用链脲酶素建立急性高血糖模型;高血糖模型雌鼠及正常雌鼠注射10IU/只孕马血清促性腺激素(PMSG)和人绒毛膜促性腺激素(hCG)超排卵,注射hCG后0h、2h、6h和10h取生发泡(GV)期卵母细胞,双光子激光扫描共焦显微镜检测GV期卵母细胞的Ca2+振荡;hCG注射后6h、10h取卵丘-卵母细胞复合体(COC),利用免疫荧光、Western blotting检测TRAIL的表达.结果 1.高血糖组卵母细胞Ca2+振荡频率高于正常对照组,于hCG注射后2h、6h差异显著(P<0.05);2.免疫荧光、Western blotting蛋白半定量检测显示,高血糖组颗粒细胞TRAIL表达量高于正常对照组,于hCG注射后6h、10h差异有显著性(P<0.05).结论 高血糖加快小鼠未成熟卵母细胞Ca2+振荡的频率;高血糖促进小鼠卵泡颗粒细胞TRAIL的表达.