线粒体损伤在镉诱导肝细胞凋亡和DNA损伤中的作用

目的探讨活性氧(reactive oxidative species, ROS)介导的线粒体损伤在镉(cadmium, Cd)暴露诱导肝L02细胞凋亡和DNA损伤中的作用。方法以肝L02细胞为研究对象,利用0~90μmol/L的Cd处理细胞24 h,采用噻唑兰法检测Cd暴露对细胞生存率的影响;以0、20和40μmol/L的Cd染毒细胞24 h,分别采用克隆形成实验、流式细胞术、彗星实验、2′,7′-二氯荧光黄双乙酸盐非标记性氧化敏感的荧光探针、线粒体红色荧光探针(Mitotracker Red CMXRos)和10-N-壬基-吖啶橙等荧光探针标记、线粒体膜电位检测试剂盒(JC-1)、ATP测定试剂盒以及Western Blot等方法,检测Cd暴露对细胞克隆形成能力、细胞凋亡、DNA损伤、ROS水平、线粒体形态、线粒体膜电位(mitochondrial membrane potential, MMP/Δψm)、线粒体质量、ATP含量及相关蛋白的影响;利用90μmol/L抗氧化剂维生素C预处理细胞1 h后给予40μmol/L Cd处理细胞24 h,检测ROS水平、Δψm、线粒体质量、ATP含量、细胞凋亡、DNA损伤及相关蛋白的改变。结果随Cd处理浓度的增高和处理时间的延长,肝L02细胞存活率显著降低,克隆形成实验结果显示,与对照组相比,20和40μmol/L Cd处理组的克隆形成率分别为8.23%和6.17%,细胞凋亡率分别为15.85%和26.26%,且B细胞淋巴瘤/白血病-2(B cell lymphoma/lewkmia-2, Bcl-2)基因的蛋白水平显著降低,Bcl-2相关X蛋白(Bcl-2 associated X protein, Bax)和激活型半胱天冬蛋白酶-3(cleaved cysteine aspastic acid-specific protease 3, cleaved-caspase-3)蛋白水平呈剂量依赖性升高,Cd处理还可诱导细胞DNA损伤的发生以及细胞内ROS的大量蓄积,伴随线粒体颗粒状改变、Δψm、线粒体质量、ATP含量和线粒体细胞色素C(cyt c)的显著降低及胞浆cyt c的表达增高(P<0.05);此外,与单独Cd处理组相比,维生素C预处理不仅能够显著增高Δψm、线粒体质量、ATP含量和线粒体cyt c,降低胞浆cyt c的表达(P<0.05),还能够减轻Cd诱导的细胞凋亡和DNA损伤。结论 Cd暴露可诱导细胞内ROS的蓄积,进而引起线粒体功能障碍,最终导致细胞凋亡和DNA损伤的发生。     

维利帕尼和多柔比星联合作用对人肝癌耐药细胞株BEL-7404增殖和凋亡的影响

目的检测PARP抑制剂维利帕尼与抗癌药物多柔比星联合应用对肝癌耐药细胞株BEL-7404增殖抑制的作用。方法以BEL-7404为研究对象,常规培养传代,经多柔比星和（或）维利帕尼处理24 h后,通过四甲基偶氮唑盐(MTT) 比色法观察细胞的增殖率变化, 采用流式细胞术Annexin V-FITC/PI双染法分析细胞凋亡水平,通过单细胞凝胶电泳实验评价DNA损伤程度,通过聚丙烯酰胺凝胶电泳（Western blotting）法检测细胞线粒体和胞浆中细胞色素c水平变化。结果多柔比星和维利帕尼联合应用组的细胞增殖率明显低于对照组和多柔比星单独处理组（P<0.01）,细胞凋亡率则显著高于对照组和多柔比星单独处理组（P<0.05）;同时,多柔比星和维利帕尼联合应用组细胞DNA损伤水平较对照组和多柔比星单独处理组亦明显加重（P<0.01）,而细胞色素C在胞浆中的水平则显著高于对照组和多柔比星单独处理组（P<0.01）。结论维利帕尼与多柔比星联合作用可以抑制BEL-7404细胞的增殖,并诱导细胞DNA损伤和凋亡,其机制与胞浆中细胞色素c的表达升高有关。     

三氧化二砷对C6胶质瘤细胞增殖和凋亡的影响

目的 探讨三氧化二砷(As2O3)对体外培养的C6胶质瘤细胞增殖和凋亡的影响.方法 以浓度为1、3、5μmol/LAs2O3分别作用于体外培养的C6胶质瘤细胞,采用甲基噻唑蓝比色法(MTT)检测细胞活性,流式细胞仪检测细胞周期和细胞凋亡,并用倒置显微镜及透射电子显微镜观察细胞形态学和超微结构的变化.结果 经1、3、5μmol/L As2O3作用后,MTT检测C6胶质瘤细胞活性受到抑制,且呈剂量时间依赖性;流式细胞仪检测凋亡细胞数目随着As2O3浓度的增加呈现逐渐递增的趋势;且上述药物干预者与未用药物干预者比较差异均有统计学意义(P＜0.05);透射电子显微镜检测药物干预者C6胶质瘤细胞较未用药物干预者体积明显缩小,可见少量空泡,细胞核不规则,核内染色质凝聚、边集,形成凋亡细胞的形态学改变等现象.结论 As2O3可抑制C6胶质瘤细胞增殖,诱导C6胶质瘤细胞凋亡.

Abstract：

Objective To investigate the effects of arsenic trioxide(As2O3) on the cell proliferation and apoptosis of C6 glioma cells in vitro.Methods The C6 glioma cells were treated by 1,3,5 μ mol/L of As2O3 with different duration and observed under the microscope and electromicroscope.The viability of C6 glioma cells was examined by methyl thiazolyl tetrazolium (MTT) assay,and cell cycle and apoptosis were examined by flow cytometry.Results After treatment of 1,3,5 μ mol/L As2O3,C6 glioma cells were inhibited obviously with a dose- and time-dependent manner (P ＜0.05) by MTT.During flow cytometry,more increasing apoptotic cells were found in different concentration As2O3.Characteristic morphological changes were observed in As2O3 intervention by transmission election microscopy including cell shrinkage,physaliphore,nuclear condensation and apoptosis and so on.Conclusion As2O3 can inhibit the cell proliferation and induce the apoptosis of C6 glioma cells.     

大豆异黄酮诱导HeLa细胞凋亡、细胞周期阻滞和[Ca~(2+)]_i浓度升高(英文)

目的探讨大豆异黄酮SI-I抑制HeLa细胞生长机制。方法 HeLa细胞经过10,20,and 40 g/ml的大豆异黄酮SI-I处理4d后,采用倒置显微镜和透射电镜观察其形态学变化,利用流式细胞仪检测细胞周期分布,通过激光扫描共聚焦显微镜观察细胞中[Ca2+]i浓度。结果倒置显微镜观察到HeLa细胞数量明显变少,细胞严重变形。透射电镜观察到HeLa细胞发生凋亡形态学变化。流式细胞仪分析发现HeLa细胞周期阻滞于G0/G1或G2/M期,细胞凋亡率随SI-I浓度的增加而上升。激光扫描共聚焦显微镜观察到凋亡的HeLa细胞内[Ca2+]i显著升高。结论大豆异黄酮SI-I抑制HeLa细胞生长是通过凋亡诱导作用,并且细胞周期阻滞和[Ca2+]i浓度升高可能是其诱导凋亡机制之一。[营养学报,2012,34(4):373-378]     

松口蘑菌丝体蛋白对HeLa细胞凋亡的影响

目的 :　研究松口蘑发酵菌丝体水提液中菌丝体蛋白质 (TMP- B)的体外抗癌作用。方法 :　体外培养 He La细胞 ,培养时添加不同浓度的 TMP- B,绘制细胞生长曲线。利用扫描电镜、流式细胞术及 DNA凝胶电泳等进行体外抗人子宫颈癌 He La细胞增殖和诱导细胞凋亡机制的研究。结果 :　 TMP- B在体外具有抑制肿瘤细胞增殖的作用。扫描电镜观察到 TMP- B处理细胞产生明显的凋亡小体 ;TMP- B对细胞周期的影响是通过抑制细胞从 S到 G2 /M期的转化来抑制He La细胞增殖 ,诱导细胞发生凋亡的 ;流式细胞仪分析图上出现凋亡峰 ;DNA电泳出现以 2 0 0 bp左右为单位的 DNA碎片。结论 :　采用液体培养方法生产的松口蘑菌丝体蛋白质 ,具有显著的抑制细胞增殖和诱导细胞凋亡作用。     

Bcl-2家族蛋白在维生素E琥珀酸酯诱导人胃癌细胞凋亡中的作用

目的:研究Bcl-2家族蛋白在维生素E琥珀酸酯(RRR-α-tocopheryl succinate,α-TOS;vitamin E succinate,VES)诱导人胃癌SGC-7901细胞凋亡线粒体途径中的作用。方法:采用噻唑蓝(MTT)法测定VES对SGC-7901细胞的半数抑制浓度(IC50值);吖啶橙/溴化乙啶(AO/EB)染色观察细胞凋亡;Mito Tracker Red CMXRos染色观察线粒体膜电位(ΔΨm)的改变;Western Blot法检测不同剂量VES对人胃癌SGC-7901细胞Bid、Bax、Bcl-2蛋白表达和细胞色素C(cytochrome C,Cyt C)蛋白表达与定位的影响。结果:VES对SGC-7901细胞的IC50值为101.45μg/ml;VES可引起SGC-7901细胞发生凋亡和线粒体膜电位下降;并引起Cyt C蛋白在细胞内重新定位、Bid蛋白剪切活化、Bax蛋白表达增加和Bcl-2蛋白表达减少。结论:VES可抑制SGC-7901细胞的生长,并通过线粒体途径诱导凋亡,其机制可能是通过剪切活化Bid蛋白、上调Bax/Bcl-2相对水平来实现的。     

槲皮素联合顺铂诱导宫颈HeLa细胞凋亡的实验研究

目的探讨槲皮素对HeLa细胞增殖和凋亡的影响以及联合顺铂的化疗效应。方法分别用不同浓度槲皮素、顺铂及不同浓度槲皮素联合顺铂处理HeLa细胞24 h;采用MTT法检测药物对HeLa细胞增殖的影响;流式细胞仪和Hochest 33258检测药物作用后的细胞凋亡情况;荧光显微镜观察用PI荧光标记的细胞核形态学变化。结果槲皮素能抑制HeLa细胞增殖,且呈剂量依赖性;槲皮素联合顺铂与相同浓度槲皮素单用及顺铂单用处理HeLa细胞相比,前者对HeLa细胞的抑制作用更显著(p<0.01);不同浓度槲皮素与顺铂联合处理HeLa细胞与槲皮素单用及顺铂单用相比,前者显著提高了HeLa细胞的凋亡率(p<0.01)。结论槲皮素联合顺铂比槲皮素单用及顺铂单用对HeLa细胞的抑制作用更显著,因而产生更强的抗肿瘤效果。     

NaHS抑制宫颈癌细胞增殖的初步机制

目的:探讨NaHS对子宫颈癌细胞增殖的影响及其机制。方法:首先将子宫颈癌HeLa细胞分成对照组和不同剂量的NaHS处理组,并采用MTT法检测细胞存活率和流式细胞仪检测细胞凋亡率。使用不同浓度的格列本脲预先处理HeLa细胞30 min,再用400μmol/L的NaHS处理24 h,探讨NaHS抑制子宫颈癌细胞增殖的初步机制。结果:不同剂量的NaHS处理组能明显抑制HeLa细胞增殖而促进HeLa细胞凋亡;格列本脲预先处理可以部分阻断这种作用。结论:NaHS抑制子宫颈癌细胞增殖可能与ATP敏感性钾通道有关。     

Dorycnium pentaphyllum Extract Has Antiproliferative Effect on Human Cervix and Colon Cancer Cells

Abstract

Although several studies have investigated the cytotoxic effects of different Fabaceae species, limited researches have been conducted on the cytotoxic effect of Dorycnium pentaphyllum. The aim of this study was to evaluate the phenolic characterization and the cytotoxic effect of D. pentaphyllum on human cervix (HeLa) and colon (WiDr) cancer cells and the possible mechanisms involved. Total phenolic content (TPC) and phenolic characterization of the extract were investigated using the Folin–Cioceltau method and RP-HPLC, respectively. The cytotoxic effect of the extract was evaluated using the MTT assay. The mechanism involved in the extract’s cytotoxic effect was then evaluated in terms of apoptosis and the cell cycle using flow cytometry, while mitochondrial membrane potential (MMP) was investigated using the fluorometric method. The TPC value of the extract was 141.2?±?0.8?mg gallic acid equivalent per g sample, and quercetin was detected as major phenolics. D. pentaphyllum extract exhibited a selective cytotoxic effect on HeLa and WiDr cells compared to normal fibroblast and colon cells, respectively. The extract induced cell cycle arrest at the S phase and apoptosis via reduced MMP in these cells. Further studies may be useful in developing a natural product based new generation pharmacological agent.     

Antiproliferative Effects of Bacillus coagulans Unique IS2 in Colon Cancer Cells

In the present study, the in vitro anticancer (antiproliferative) effects of Bacillus coagulans Unique IS2 were evaluated on human colon cancer (COLO 205), cervical cancer (HeLa), and chronic myeloid leukemia (K562) cell lines with a human embryonic kidney cell line (HEK 293T) as noncancerous control cells. The Cytotoxicity assay (MTT) clearly demonstrated a 22%, 31.7%, and 19.5% decrease in cell proliferation of COLO 205, HeLa, and K562 cells, respectively, when compared to the noncancerous HEK 293T cells. Normal phase-contrast microscopic images clearly suggested that the mechanism of cell death is by apoptosis. To further confirm the induction of apoptosis by Unique IS2, the sub-G0–G1 peak of the cell cycle was quantified using a flow cytometer and the data indicated 40% of the apoptotic cells in Unique IS2-treated COLO cells when compared with their untreated control cells. The Western blot analysis showed an increase in pro-apoptotic protein BAX, decrease in antiapoptotic protein, Bcl2, decrease in mitochondrial membrane potential, increase in cytochrome c release, increase in Caspase 3 activity, and cleavage of poly(ADP-ribose) polymerase. The present study suggests that the heat-killed culture supernatant of B. coagulans can be more effective in inducing apoptosis of colon cancer cells and that can be considered for adjuvant therapy in the treatment of colon carcinoma.     

2,5-hexanedione induced apoptosis in mesenchymal stem cells from rat bone marrow via mitochondria-dependent caspase-3 pathway

2,5-hexanedione (HD) induces apoptosis of nerve cells. However,the mechanism of HD-induced apoptosis remains unknown. Mesenchymal stem cells (MSCs) are multipotential stem cells with the ability to differentiate into various cell types. This study is designed to investigate the apoptosis induced by HD in rat bone marrow MSCs (BMSCs) and the related underlying mechanisms. The fifth generation of MSCs was treated with 0, 10, 20 and 40 mM HD respectively. The viability of BMSCs was observed by MTT. Apoptosis were estimated by Hoechst 33342 staining and TUNEL assay. The disruption of mitochondrial transmembrane potential (MMP) was examined by JC-1 staining. Moreover, the expression of Bax and Bcl-2, cytochrome c release, and caspase-3 activity were determined by real-time RT-PCR, Western blot and Spectrophotometry. Our results showed that HD induced apoptosis in MSCs in a dose dependent manner. Moreover, HD downregulated the Bcl-2 expression,upregulated the Bax expression and the Bax/Bcl-2 ratio, promoted the disruption of MMP, induced the release of cytochrome c from mitochondria to cytosol, and increased the activity of caspase-3 in MSCs. These results indicate that HD induces apoptosis in MSCs and the activated mitochondria-dependent caspase-3 pathway may be involved in the HD-induced apoptosis.     

Induction of apoptosis signaling by glycoprotein of Capsosiphon fulvescens in human gastric cancer (AGS) cells

Capsosiphon fulvescens is a well-known green sea algae that has been touted in recent years as a potential anticancer drug. In this study, C. fulvescens glycoprotein (Cf-GP) showed proapoptotic signaling in AGS cells. An MTS assay indicated that Cf-GP inhibited the proliferation of AGS cell lines in a dose-dependent manner. Cells were treated with Cf-GP and the expression of proteins associated with apoptosis was examined by Western blotting. Based on the Western blot, expression of Cf-GP-activated caspase-cascade and PARP, which is a substrate of caspase-3 and -8, and proteins of the Bcl-2 family was observed. Cf-GP treatment stimulated the release of cytochrome C and apoptotic protease activating factor-1 from mitochondria to the cytosol. Cf-GP inhibited the growth of AGS cells through induction of sub-G1 phase arrest. We confirmed that sub-G1-phase arrest was associated with a decrease in the expression of cyclin D, cyclin E, Cdk2, Cdk4, and Cdk6, and an increase in the protein levels of p21 and p27. As a result, the increased sub-G1 ratio appears to be inhibited by cell proliferation. Therefore, we can confirm apoptosis in the AGS cells. Our results suggest that Cf-GP could be a potential source of biofunctional material that shows anticancer effects in human gastrointestinal cancer.     

二十碳五烯酸（EPA）对胃癌细胞增殖与凋亡的影响

目的观察二十碳五烯酸（EPA）对人胃癌细胞系增殖与凋亡的影响,并探讨其作用机制。方法以终浓度为10、20、40μg／ml的EPA作用于SGC-7901和MGC-803人胃癌细胞系24-72h。采用四甲基偶氮唑蓝法检测细胞增殖抑制率,采用流式细胞技术分析细胞周期分布与细胞凋亡,利用荧光探针rhodamine 123测定线粒体膜电位,酶联免疫吸附法测定线粒体和胞浆中细胞色素C水平,荧光光谱法测定凋亡效应酶半胱天冬蛋白酶-3（caspase-3）活性。结果经10-40μg／ml EPA处理的胃癌细胞增殖率显著降低,且呈现时间依赖性。与对照组比较,经40g／ml EPA作用72h后,SGC-7901和MGC-803细胞G0／G1期细胞的比例均增加（P=0.006、P=0.009）。经40g／ml EPA作用24h后,胃癌细胞线粒体膜电位显著低于对照组（P=0.001、P=0.047）;线粒体内细胞色素C的含量明显少于对照组（P=0.001、P=0.000）,而细胞浆中细胞色素C含量显著高于对照组（P=0．001、P=0．000）。在SGC-7901细胞中,caspase-3活性随着EPA（40g／mL）作用时间延长而升高。结论EPA通过诱导细胞周期阻滞和激活线粒体通路,抑制人胃癌细胞增殖,诱导细胞凋亡。     

Latex of Euphorbia Antiquorum Induces Apoptosis in Human Cervical Cancer Cells via c-Jun N-terminal Kinase Activation and Reactive Oxygen Species Production

Latex of Euphorbia antiquorum (EA) has inhibitory effects on several different cancer cell lines. However, the molecular mechanism of EA inhibitory effects on human cervical cancer HeLa cell growth has not been explored. EA induced apoptosis, which was characterized by morphological change, DNA fragmentation, increased sub-G1 population, and alterations in levels of apoptosis-associated proteins. Treatment with EA increased cell death and expression levels of caspase-8, -9, and -3. EA suppressed expression of Bcl-2, increased Bax, and reduced cleavage of Bid and the translocation of tBid to the mitochondria and the release of cytochrome c from mitochondria. EA caused a loss of mitochondrial membrane potential (ΔΨm) and an increase in cellular reactive oxygen species (ROS). EA-induced ROS formation was suppressed by cyclosporine A (an inhibitor of the ΔΨm) or allopurinol (an effective scavenger of ROS). EA also increased expression of Fas, FasL, and c-Jun N-terminal kinase (JNK), p38, and mitogen-activated protein kinase (MAPK) and decreased expression of extracellular signal-regulated kinase (ERK) 1/2-p. Co-treatment with the JNK inhibitor SP600125 inhibited EA-induced apoptosis and the activation of caspase-8, -9, and -3. Results of this study provide support for the hypothesis that EA causes cell death via apoptotic pathways in human cervical adenocarcinoma HeLa cells.     

Latex of Euphorbia antiquorum induces apoptosis in human cervical cancer cells via c-jun n-terminal kinase activation and reactive oxygen species production

Latex of Euphorbia antiquorum (EA) has inhibitory effects on several different cancer cell lines. However, the molecular mechanism of EA inhibitory effects on human cervical cancer HeLa cell growth has not been explored. EA induced apoptosis, which was characterized by morphological change, DNA fragmentation, increased sub-G1 population, and alterations in levels of apoptosis-associated proteins. Treatment with EA increased cell death and expression levels of caspase-8, -9, and -3. EA suppressed expression of Bcl-2, increased Bax, and reduced cleavage of Bid and the translocation of tBid to the mitochondria and the release of cytochrome c from mitochondria. EA caused a loss of mitochondrial membrane potential (ΔΨm) and an increase in cellular reactive oxygen species (ROS). EA-induced ROS formation was suppressed by cyclosporine A (an inhibitor of the ΔΨm) or allopurinol (an effective scavenger of ROS). EA also increased expression of Fas, FasL, and c-Jun N-terminal kinase (JNK), p38, and mitogen-activated protein kinase (MAPK) and decreased expression of extracellular signal-regulated kinase (ERK) 1/2-p. Co-treatment with the JNK inhibitor SP600125 inhibited EA-induced apoptosis and the activation of caspase-8, -9, and -3. Results of this study provide support for the hypothesis that EA causes cell death via apoptotic pathways in human cervical adenocarcinoma HeLa cells.     

白英提取物单独或与顺铂联合对宫颈癌Hela细胞的影响

目的:观察中药白英提取物(Solanum lyratum Thunb extract,STE)对体外培养宫颈癌HeLa细胞的作用,以及STE对HeLa细胞对顺铂(DDP)的敏感性的影响。方法:体外培养宫颈癌HeLa细胞分别用STE、DDP、DDP+STE处理细胞,MTT法检测细胞活性,流式细胞术检测细胞凋亡。结果:白英提取物及DDP对宫颈癌HeLa细胞的生长均表现出明显的抑制作用,低剂量STE可加强DDP对HeLa细胞的抑制作用;流式细胞术检测结果显示,白英提取物作用后HeLa细胞出现凋亡,且随着时间的延长,凋亡细胞的比例明显增加。结论:白英提取物对宫颈癌HeLa细胞有明显的抑制生长和促凋亡作用,同时能提高HeLa细胞对顺铂的敏感性。     

飞燕草素葡萄糖苷抑制人血管内皮细胞凋亡的作用及机制研究

目的观察飞燕草素葡萄糖苷(Dp)对氧化低密度脂蛋白(oxLDL)诱导的人血管内皮细胞株EA.hy926凋亡的影响,并探索相关分子机制。方法 CCK-8等试剂盒分别检测不同浓度的Dp(25、50、100、200μmol/L)对oxLDL诱导的细胞活力、MDA和LDH生成的影响;激光共聚焦显微镜检测Dp对oxLDL诱导的细胞内ROS生成、线粒体膜电位和线粒体膜通道孔开放状态的影响;流式细胞仪检测不同浓度的Dp对oxLDL诱导的细胞凋亡的影响;Western blot法检测Dp对oxLDL诱导的bcl-2及bax蛋白表达水平的影响。结果 Dp能明显抑制oxLDL诱导的EA.hy926细胞活力降低,及MDA和LDH生成增加,并呈浓度依赖关系;与oxLDL处理组比较,不同浓度的Dp预处理细胞2 h后,总凋亡细胞百分比显著降低(P<0.05);Dp能明显抑制oxLDL诱导的细胞内ROS生成、线粒体膜电位下降和线粒体膜通道孔开放,并能明显抑制oxLDL诱导的细胞内bcl-2蛋白表达水平降低及bax蛋白表达水平增高。结论 Dp可能通过线粒体途径抑制oxLDL诱导的血管内皮细胞凋亡发生。     

奈达铂对宫颈癌Hela及Siha细胞的作用

目的:比较奈达铂(NDP)体外对人宫颈腺癌细胞Hela细胞及鳞癌细胞Siha的抑制作用并探讨其作用机制。方法:以2.5～40μg/ml NDP分别作用于Hela及Siha细胞24 h,48 h,72 h,MTT法检测抑制率,计算半数抑制浓度(IC50);利用流式细胞术(FCM)观察细胞凋亡率及周期变化;半定量PCR法分析基因半胱氨酸天门冬氨酸蛋白酶3(Caspase 3,Caspase 9)及基质金属蛋白酶2(MMP-2)的表达变化。结果:NDP对宫颈癌Hela细胞及Siha细胞均有抑制作用,呈时间-剂量依赖性,NDP对Hela细胞的抑制率大于Siha细胞;FCM显示,随时间、浓度增加,Hela及Siha细胞凋亡率增加,相同浓度和作用时间下,Hela细胞的凋亡率大于Siha细胞(P<0.05),S期细胞的比例增加,G0/G1期比例减少,G2/M期比例变化不明显;与对照组相比,Caspase 3,Caspase 9表达增加,MMP-2表达减少。结论:NDP可抑制宫颈腺癌细胞Hela及鳞癌细胞Si-ha的增殖,NDP体外对Hela细胞的抑制作用更强。NDP的作用机制与诱导细胞凋亡、细胞周期阻滞、上调Caspase3和Caspase9的表达有关;下调MMP-2的表达可能有利于降低宫颈癌细胞的侵袭力。     

L-Threonine induces heat shock protein expression and decreases apoptosis in heat-stressed intestinal epithelial cells

ObjectivesOsmotically acting amino acids can be cytoprotective following injury. As threonine (THR) induces osmotic cell swelling, our aim was to investigate the potential for THR to induce cellular protection in intestinal epithelial cells and evaluate possible mechanisms of protection.MethodsCells treated with a range of THR doses were evaluated following heat stress (HS) injury. Alpha-aminoisobutyric acid (AIB), a non-metabolizable amino acid analog, was used as an osmotic control. MTS assays were used to assess cell survival. Heat shock protein (HSP) expression and cleaved caspase-3 (CC3) were evaluated via Western blot. Cell morphology and cell size were analyzed via microscopy.ResultFollowing HS, THR treatment increased cell viability in a dose dependent manner vs. non-THR treated cells (CT). The non-metabolized amino acid analogue, AIB, also increased cell survival in heat-stressed cells versus HS controls. HSP70 and HSP25 expression increased with THR and AIB treatment versus HS controls. THR also increased HSP25 in non-stressed cells. Microscopic evaluation revealed both THR and AIB preserved the structural integrity of the actin cytoskeleton in heat-stressed cells versus HS controls. THR, but not AIB, enhanced nuclear translocation of HSP25 during HS. This nuclear translocation was associated with a 60% decrease in apoptosis in heat-stressed cells with THR. No antiapoptotic effect was observed with AIB.ConclusionsThis is the first demonstration that THR increases HSP70 and HSP 25 and protects cells from HS. THR's mechanism of protection may involve cytoskeletal stabilization, HSP up-regulation and nuclear translocation, and decreased apoptosis. THR's protection appears to involve both cell-swelling–dependent and –independent processes.     

舒林酸抑制结肠癌细胞株增殖及其转移机制的研究

目的 研究非选择性COX抑制荆舒林酸对结肠癌细胞株HT-29生长的影响及其参与引起细胞死亡的可能机制.方法 采用四甲基偶氮唑蓝(MTT)比色法检测舒林酸对结肠癌细胞增殖的抑制,激光共聚焦显微镜(LSM)及荧光显微镜观察细胞凋亡,流式细胞仪(FCM)分析其对细胞凋亡及细胞周期的影响.结果 MTT显示舒林酸能呈剂量和浓度依赖性抑制HT-29的增殖.TUNEL染色荧光显微镜下观察凋亡细胞呈棕褐色,AnnexinV/PI染色后LSM观察凋亡细胞胞膜绿染,胞核红染或呈桔黄色,FCM显示此药促进细胞的凋亡,使处于G0/G1期的细胞比例显著降低.结论 舒林酸可抑制结肠癌细胞株HT-29生长,促进其凋亡,其机制可能与其阻止细胞周期的进展有关.