硫化氢抑制葡萄糖调节蛋白78减轻6-OHDA诱导的PC12细胞损伤

目的：探讨硫化氢（H2S）是否通过改变葡萄糖调节蛋白78（GRP78）的表达参与其对抗6-羟基多巴胺（6-OHDA）诱导PC12细胞损伤的保护作用。方法应用具有神经毒性的6-OHDA损伤PC12细胞为帕金森病细胞模型,以硫氢化钠（NaHS）作为H2S的供体;应用CCK-8比色法检测细胞存活率;DFCH-DA染色检测细胞内活性氧（ROS）的水平;Rh123染色检测细胞线粒体膜电位（MMP）;Western blot检测GRP78的表达。结果200μmol/L的6-OHDA引起PC12细胞的存活率显著降低,ROS生成增加及MMP降低,且诱导了GRP78的高表达。应用25~400μmol/L的NaHS预处理30 min,呈浓度依赖性抑制6-OHDA引起的细胞存活率降低,其中400μmol/L的NaHS作用最明显,此浓度也可以显著减少6-OHDA引起的ROS增多,提高MMP,同时明显抑制6-OHDA诱导的GRP78高表达。结论 H2S具有抗6-OHDA氧化应激损伤的PC12细胞保护作用,抑制内质网应激分子GRP78的表达可能是其机制之一。     

竹叶花椒提取物诱导BRL 3A细胞产生DNA损伤和凋亡的机制研究

目的 探讨竹叶花椒甲醇提取物(ZADCM)对大鼠肝细胞BRL 3A的DNA损伤和凋亡的潜在作用机制。方法 体外培养大鼠正常肝细胞(BRL 3A),CCK-8法测量细胞毒性,荧光探针测定ROS变化,微核实验检测细胞染色体损伤,彗星实验检测DNA损伤,流式细胞术测定细胞凋亡情况,Western blot测定DNA损伤相关蛋白以及凋亡相关蛋白表达量。结果 随着ZADCM浓度增加,细胞存活率下降,具有时间-剂量关系;ZADCM可以剂量依赖方式诱导细胞内ROS累积,导致染色体损伤与DNA损伤,表现为微核细胞率、细胞尾长、尾部DNA含量、OTM值以及γH2AX、p-ATM和p-Chk2表达量升高;并进一步诱导细胞凋亡。结论 竹叶花椒甲醇提取物可导致细胞存活率下降,直接诱导细胞内ROS升高,进一步引起细胞染色体损伤和ATM介导的DNA损伤,最终导致细胞凋亡。     

天麻素通过抑制细胞凋亡保护谷氨酸诱导的PC12细胞损伤

目的：研究天麻素（Gastrodin）对谷氨酸诱导的大鼠肾上腺嗜铬细胞瘤PC12细胞损伤的影响及可能机制。方法：以谷氨酸建立体外培养PC12细胞损伤模型并采用MTT比色法测定细胞存活率;AO/EB双染法经荧光显微镜观察细胞凋亡形态;采用流式细胞术检测细胞内活性氧含量以及Annexin V/PI染色后的细胞凋亡率;Western blot法检测细胞内Caspase-3蛋白表达。结果：天麻素可明显抑制谷氨酸诱导的PC12细胞凋亡,在0.1～10μmol/L剂量呈一定的量效关系;同时,天麻素可明显抑制谷氨酸引起的活性氧（ROS）的累积,降低谷氨酸诱导的活性Caspase-3蛋白的表达,降低PC12细胞的凋亡率,在0.1～10μmol/L剂量呈量效相关性。结论：在一定剂量范围内,天麻素对谷氨酸损伤的PC12细胞具有保护作用,其机制可能与减少ROS的生成,阻止氧化损伤的发生,抑制Caspase-3途径依赖的细胞凋亡相关。     

咖啡酸苯乙酯通过Nrf-2途径抑制地塞米松诱导的成骨细胞凋亡

目的 探讨Nrf-2途径在咖啡酸苯乙酯(CAPE)抑制地塞米松(DEX)诱导成骨细胞凋亡中的作用.方法 用细胞贴壁法培养小鼠颅顶前骨细胞(MC3T3-E1),采用10μmol/L DEX建立细胞损伤模型,以不同浓度CAPE(0.05、0.25、1μmol/L)预处理细胞2h后加入地塞米松共孵育24h;细胞增殖-毒性检测试剂盒(CCK-8)检测细胞增殖活性;依据CCK-8检测结果确定药物浓度后将实验分为对照组、咖啡酸苯乙酯组(CAPE组)、地塞米松组(DEX组),咖啡酸苯乙酯+地塞米松组(CAPE+DEX组);DCFH-DA荧光探针检测细胞内活性氧(reactive oxygen species,ROS)水平;Caspase-3活性检测试剂盒检测Caspase-3酶活性,Western blot法检测Nrf-2 途径相关蛋白Nrf-2和血红素氧合酶1(heme oxygenase,HO-1)蛋白表达水平;流式细胞仪检测细胞凋亡率.结果 10μmol/L DEX作用下细胞形态发生明显变化,损伤作用明显.与对照组相比,DEX组内细胞存活率显著下降(P＜0.01),而细胞内 ROS水平、细胞凋亡率及Caspase-3酶活性显著增加(P＜0.01);同时,Nrf-2途径相关蛋白 Nrf-2及HO-1表达量减少,差异有统计学意义(P＜0.01).与DEX组相比,CAPE+DEX组细胞存活率显著上升(P＜0.01),Nrf-2途径相关蛋白 Nrf-2及HO-1表达明显增加(P＜0.01);此外,CAPE+DEX组细胞内 ROS 水平、细胞凋亡率及Caspase-3酶活显著降低,差异有统计学意义(P＜0.01).结论 咖啡酸苯乙酯可以通过Nrf-2途径降低细胞内ROS水平进而减轻地塞米松诱导的氧化应激所致细胞损伤及凋亡.     

黄芩苷对香烟烟雾提取物诱导的细胞损伤保护作用研究

目的研究黄芩苷对香烟烟雾提取物（cigarette smoke extract,CSE）诱导的人肺泡上皮细胞损伤的影响,并初步探讨其机制。方法制备CSE,用不同浓度黄芩苷对人肺泡上皮细胞A549进行预处理,再用CSE刺激细胞。用MTT法检测细胞存活率,流式细胞术测定细胞凋亡率,彗星实验观察DNA损伤情况,荧光法测定细胞内活性氧（reactive oxygen species,ROS）含量。结果随着CSE浓度的增加和作用时间的延长,细胞存活率下降,各组之间有统计学差异（P〈0.05）;黄芩苷能减少CSE诱导的细胞存活率下降,抑制CSE诱导的细胞内ROS的产生,并有剂量依赖关系（P〈0.05或P〈0.01）;黄芩苷＋CSE组细胞的彗星尾长、尾部DNA含量、尾距、Olive尾距均小于CSE组（P〈0.05）,并且黄芩苷可以减少CSE导致的细胞凋亡。结论黄芩苷可以拮抗CSE对细胞的损伤,提高细胞的存活率,降低凋亡率,其原因可能与黄芩苷能降低细胞ROS含量,减少DNA损伤有关。     

原花青素对H2O2引起PC12细胞的内质网应激损伤的保护作用

目的：研究H2O2诱导PC12细胞的内质网应激损伤作用及原花青素对其预防作用。方法：H2O2 100μmol/L 诱导PC12细胞损伤18小时复制阿尔茨海默病模型（AD）,原花青素40mg/L、SB203580 10μmol/L、Tempol 800μmol/L于造模前30min预处理后,RT-PCR检测BiP/GRP-78 mRNA水平变化;流式细胞术检测细胞内反应氧产物（ROS）阳性细胞生成率,计算平均荧光强度。结果：H2O2 100μmol/L 诱导PC12细胞损伤后BiP/GRP-78 mRNA表达显著增高,ROS生成增加;原花青素40mg/L可显著降低BiP/GRP-78 mRNA水平（P〈0.01）,减少细胞内ROS生成（P〈0.01）。结论：原花青素可减轻H2O2引起PC12细胞的内质网应激损伤。     

新型番荔枝酰胺衍生物FLZ对去血清培养损伤星形胶质细胞的保护作用

目的评价番荔枝酰胺衍生物FLZ对去血清培养损伤星形胶质细胞的作用,并探讨其可能的作用机制。方法在去血清培养的星形胶质细胞模型上,以3-（4,5-二甲基噻唑-2）-2,5-二苯基四氮唑溴盐（MTT）法检测细胞存活率,Hochest 33342染色观察细胞核形态变化,荧光比色法检测细胞内活性氧簇（ROS）生成。酶联免疫吸附测定（ELISA）方法检测细胞培养液中S100B蛋白含量。生化法检测超氧化物歧化酶（SOD）和谷胱甘肽过氧化物酶（GSH-Px）活力及丙二醛（MDA）与谷胱甘肽（GSH）含量。结果去血清培养损伤星形胶质细胞使其存活率下降,细胞内ROS生成增多,S100B分泌增加,细胞内SOD活力下降,GSH-Px活力升高,MDA生成增多,GSH含量减少。FLZ能提高星形胶质细胞在去血清培养状态下的存活率,减少去血清培养引起的S100B过度分泌,减少细胞内ROS和MDA过度生成,减少GSH含量,抑制SOD活力下降,降低GSH-Px活力过度升高,从而降低去血清培养对星形胶质细胞引起的氧化应激损伤。结论 FLZ对星形胶质细胞去血清培养损伤有明显的保护作用,这种保护作用可能与降低去血清培养损伤引起的S100B分泌增加、减少细胞内ROS的生成及增强细胞自身抗氧化系统相关。     

五味子酚对氧化低密度脂蛋白引起的血管内皮细胞及神经细胞损伤的保护作用

目的：氧化低密度脂蛋白（ox-LDL）对血管内皮细胞及神经细胞均有毒性作用，与动脉粥样硬化，神经退行性疾病的发展密切相关。本文探讨了具有很强抗氧化作用的五味子有效成分五味子酚（Sal）对ox-LDI。引起的牛主动脉内皮细胞及神经细胞NGl08-15的损伤和凋亡的保护作用及其初步的机制。方法：以细胞形态，MTT法，LDH释放检测ox-LDL对细胞的损伤及Sal的作用；DNA电泳，核染色质荧光染色观察细胞的凋亡，流式细胞术检测凋亡细胞的百分率；自由基（ROS）荧光探针DCF检测细胞内ROS变化，Western-blot法检测线粒体细胞色素C的释放。结果：ox-LDL可引起内皮细胞和NGl08-15细胞的损伤、凋亡，细胞形态发生明显变化，存活率降低，胞内LDH泄漏增加，出现核染色质聚集、DNA梯状条带，流式细胞术检测出现G1亚峰，凋亡细胞百分率显著增加，并可刺激细胞内ROS产生，促进NGl08-15细胞线粒体细胞色素C的释放增加。提前加入Sal对上述细胞损伤具有明显的保护作用，并能显著抑制细胞的凋亡，减少细胞内ROS产生，降低细胞色素C的表达。结论：Sal能抑制ox-LDL引起的血管内皮细胞及神经细胞的毒性损伤、凋亡，其机制可能与减少胞内自由基、抑制线粒体细胞色素C的释放有关。     

伊拉地平对 MPP＋损伤 PC12细胞保护作用的实验研究

目的：研究伊拉地平（ ISR ）对1-甲基-4-苯基吡啶离子（ MPP＋）损伤的PC12细胞的保护作用及可能机制。方法MPP ＋处理PC12细胞建立帕金森病细胞模型;4-甲基偶氮唑蓝（ MTT）比色法检测细胞存活率;双氯荧光黄乙酸乙酯（ DCFH-DA）染色流式细胞术检测细胞内活性氧（ ROS）的生成;JC-1染色流式细胞术检测细胞线粒体膜电位（ MMP）。结果1 mmol · L－1MPP＋处理PC12细胞24 h后能明显抑制细胞生长（P＜0．01）;降低线粒体膜电位;ROS含量增加。2μmol· L－1伊拉地平预处理后, PC12细胞存活率显著增加（ P＜0．01）;线粒体膜电位升高;ROS生成减少。结论伊拉地平对MPP＋损伤的PC12细胞具有保护作用,其作用机制可能与维持线粒体正常膜电位,稳定线粒体功能,阻止线粒体氧化应激发生有关。     

脂质立方液晶设计制备与表征评价研究进展

目的：研究伊拉地平（ ISR ）对1-甲基-4-苯基吡啶离子（ MPP＋）损伤的PC12细胞的保护作用及可能机制。方法MPP ＋处理PC12细胞建立帕金森病细胞模型;4-甲基偶氮唑蓝（ MTT）比色法检测细胞存活率;双氯荧光黄乙酸乙酯（ DCFH-DA）染色流式细胞术检测细胞内活性氧（ ROS）的生成;JC-1染色流式细胞术检测细胞线粒体膜电位（ MMP）。结果1 mmol · L－1MPP＋处理PC12细胞24 h后能明显抑制细胞生长（P＜0．01）;降低线粒体膜电位;ROS含量增加。2μmol· L－1伊拉地平预处理后, PC12细胞存活率显著增加（ P＜0．01）;线粒体膜电位升高;ROS生成减少。结论伊拉地平对MPP＋损伤的PC12细胞具有保护作用,其作用机制可能与维持线粒体正常膜电位,稳定线粒体功能,阻止线粒体氧化应激发生有关。     

Size-dependent cytotoxicity of amorphous silica nanoparticles in human hepatoma HepG2 cells

The purpose of this study is to compare the potential cytotoxicity induced by amorphous silica particles with different sizes. The effects of one fine particle (498 nm) and three nanoparticles (68, 43, and 19 nm) on cultured human hepatoma (HepG2) cells were investigated by detecting morphological changes, cell viability, cytomembrane integrity, DNA damage, cell cycle distribution, and apoptosis after the cells were treated with 100 μg/mL of four silica particles for 24 h. The results indicated that in HepG2 cells, the cytotoxicity generated by silica particles strongly depended on the particle size, and smaller silica particle possessed higher toxic effect. In order to further elucidate the possible mechanisms of cell injuries, intracellular reactive oxygen species (ROS) was measured. Increased ROS level was also observed in a size dependent way. However, the result showed the fine particle did not promote intracellular ROS level significantly, while cell injuries were detected in this treated group. Thus, our data demonstrated that exposure to different sizes of silica particles resulted in a size dependent cytotoxicity in cultured HepG2 cells, and ROS generation should be one possible damage pathway but might not be completely responsible for the toxic effect produced by silica particles.     

Atypical apoptosis in L929 cells induced by evodiamine isolated from Evodia rutaecarpa

Two alkaloids, evodiamine and rutaecarpine, isolated from the dried fruits of Evodia rutaecarpa Bentham were evaluated in vitro for antiproliferation activity on tumor cells versus human peripheral blood mononuclear cells (PBMC). Evodiamine had more potent cytotoxic effects on five tumor cell lines (human malignant melanoma A375-S2, human cervical cancer HeLa, human breast adenocarcinoma MCF7, human acute monocytic leukemia THP-1, murine fibrosarcoma L929) than rutaecarpine. Moreover, evodiamine did not affect PBMC viability for a 36 h culture period. Although apoptotic bodies were observed in evodiamine-treated L929 cells stained with Hoechst 33258, DNA fragmentation as a hallmark of apoptosis was not found. Caspases were involved in the protection of L929 cells against cell death. Evodiamine initiated atypical apoptosis in L929 cells by cycle arrest at the G0/G1 phase.     

Increase the cisplatin cytotoxicity and cisplatin-induced DNA damage in HepG2 cells by XRCC1 abrogation related mechanisms

Cisplatin is one of the most potent chemotherapeutic anticancer drugs for the treatment of various cancers. The cytotoxic action of the drug is often thought to be associated with its ability to bind DNA to form cisplatin–DNA adducts. Impaired DNA repair processes including base excision repair (BER) play important roles on its cytotoxicity. XRCC1 is a key protein known to play a central role at an early stage in the BER pathway. However, whether XRCC1 contributes to decrease the cisplatin cytotoxicity and cisplatin-induced DNA damage in HepG2 still remains unknown. Hence, the purpose of this study was to explore whether abrogation of XRCC1 gene expression by short hairpin RNAs (shRNA) could reduce DNA repair and thus sensitize liver cancer cells to cisplatin. We abrogated the XRCC1 gene in HepG2 cell using shRNA transfection. Cell viability was measured by MTT assay and clonogenicity assay. Comet assay was used to detect the DNA damage induced by cisplatin. The host cell reactivation was employed to assess the DNA repair capacity of cisplatin-damaged luciferase reporter plasmid. Flow cytometry analysis was used to determine cisplatin-induced apoptosis, cell cycle and reactive oxygen species (ROS). The results showed that abrogation of XRCC1 could sensitize HepG2 cells to cisplatin. This enhanced cytotoxicity could be attributed to the increased DNA damage and reduced DNA repair capacity. Increasing cell cycle arrest and intracellular ROS production lead to more tumor cell apoptosis and then enhanced the cisplatin cytotoxicity. Our results suggested that the cisplatin cytotoxicity may increase by targeting inhibition of XRCC1.     

Atypical apoptosis in L929 cells induced by evodiamine isolated from Evodia rutaecarpa

Two alkaloids, evodiamine and rutaecarpine, isolated from the dried fruits of Evodia rutaecarpa Bentham were evaluated in vitro for antiproliferation activity on tumor cells versus human peripheral blood mononuclear cells (PBMC). Evodiamine had more potent cytotoxic effects on five tumor cell lines (human malignant melanoma A375-S2, human cervical cancer HeLa, human breast adenocarcinoma MCF7, human acute monocytic leukemia THP-1, murine fibrosarcoma L929) than rutaecarpine. Moreover, evodiamine did not affect PBMC viability for a 36?h culture period. Although apoptotic bodies were observed in evodiamine-treated L929 cells stained with Hoechst 33258, DNA fragmentation as a hallmark of apoptosis was not found. Caspases were involved in the protection of L929 cells against cell death. Evodiamine initiated atypical apoptosis in L929 cells by cycle arrest at the G0/G1 phase.     

Titanium dioxide nanoparticles induced cytotoxicity, oxidative stress and DNA damage in human amnion epithelial (WISH) cells

Titanium dioxide nanoparticles (TiO(2)-NPs) induced cytotoxicity and DNA damage have been investigated using human amnion epithelial (WISH) cells, as an in vitro model for nanotoxicity assessment. Crystalline, polyhedral rutile TiO(2)-NPs were synthesized and characterized using X-ray diffraction (XRD), UV-Visible spectroscopy, Fourier transform infra red (FTIR) spectroscopy, and transmission electron microscopic (TEM) analyses. The neutral red uptake (NRU) and [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] (MTT) assays revealed the concentration dependent cytotoxic effects of TiO(2)-NPs (30.6nm) in concentration range of 0.625-10μg/ml. Cells exposed to TiO(2)-NPs (10μg/ml) exhibited significant reduction (46.3% and 34.6%; p<0.05) in catalase activity and glutathione (GSH) level, respectively. Treated cells showed 1.87-fold increase in intracellular reactive oxygen species (ROS) generation and 7.3% (p<0.01) increase in G(2)/M cell cycle arrest, as compared to the untreated control. TiO(2)-NPs treated cells also demonstrated the formation of DNA double strand breaks with 14.6-fold (p<0.05) increase in Olive tail moment (OTM) value at 20μg/ml concentration, vis-à-vis untreated control, under neutral comet assay conditions. Thus, the reduction in cell viability, morphological alterations, compromised antioxidant system, intracellular ROS production, and significant DNA damage in TiO(2)-NPs exposed cells signify the potential of these NPs to induce cyto- and genotoxicity in cultured WISH cells.     

Neoplastic cell response to tiopronin-coated gold nanoparticles

The present study characterized the in vitro biological response of a comprehensive set of cancer cell lines to gold nanoparticles (2.7 nm) coated with tiopronin (AuNPs-TP). Our findings suggest that upon entering cells, the AuNPs-TP are sequestered in vacuoles such as endosomes and lysosomes, and mostly localize in perinuclear areas. Peak cell accumulation was achieved at 8 hours after incubation. L929 and H520 cells showed more than 75% surviving fraction when treated with 0.5 mg/mL of AuNPs-TP for 24 hours, whereas the surviving fractions were 60% in MCF-7 and 20% in HeLa cells. Reactive oxygen species (ROS) production by the AuNPs-TP was dependent on cell line and exposure time. Antioxidants inhibited ROS generation to various extents, with glutathione and tiopronin being most effective. Overall, exposure time, concentration of the AuNPs-TP, and cell line influenced neoplastic cell response. Furthermore, the mechanism of cytotoxicity of the AuNPs-TP was found to be ROS generation.From the Clinical EditorThis study describes the basic intracellular characteristics of Tiopronin-Au nanoparticles from the standpoint of their anti-cancer activity in different cancer cell cultures.     

Suppressing growth,migration, and invasion of human hepatocellular carcinoma HepG2 cells by Catharanthus roseus‑silver nanoparticles

Application of silver nanoparticles serves as a new approach in cancer treatment due to its unique features. Biosynthesis of silver nanoparticles using plant is advantageous since they are easily accessible, nontoxic and produce quicker reaction compared to other methods. To evaluate the cytotoxicity, mechanism of cell death and DNA damage of biosynthesized Catharanthus roseus-silver nanoparticles on human liver cancer (HepG2) cells. The antiproliferative activity of Catharanthus roseus‑silver nanoparticles was measured using MTT assay. The cytotoxic effects were further evaluated by measuring nitric oxide and reactive oxygen species (ROS). The mechanism of cell death was determined by annexin-FITC/propidium iodide, mitochondrial membrane potential (MMP) and cell cycle assays. The assessment of DNA damage was evaluated using Comet assay method. The uptake of the nanoparticles were evaluated by Transmission Electron Microscopy (TEM). Catharanthus roseus‑silver nanoparticles has inhibited the proliferation of HepG2 cells in a time-dependent manner with a median IC₅₀ value of 3.871 ± 0.18 μg/mL. The concentration of nitrite and ROS were significantly higher than control. The cell death was due to apoptosis associated with MMP loss, cell cycle arrest, and extensive DNA damage. TEM analysis indicated the presence of free nanoparticles and endosomes containing the nanoparticles. The findings show that Catharanthus roseus‑silver nanoparticles have produced cytotoxic effects on HepG2 cells and thus may have a potential to be used as an anticancer treatment, particularly for hepatocellular carcinoma.     

Curcumin attenuates quinocetone-induced oxidative stress and genotoxicity in human hepatocyte L02 cells

Abstract

Quinocetone (QCT), a new quinoxaline 1,4-dioxides, has been used as antimicrobial feed additive in China. Potential genotoxicity of QCT was concerned as a public health problem. This study aimed to investigate the protective effect of curcumin on QCT-induced oxidative stress and genotoxicity in human hepatocyte L02 cells. Cell viability and intracellular reactive oxygen species (ROS), biomarkers of oxidative stress including superoxide dismutase (SOD) activity and glutathione (GSH) level were measured. Meanwhile, comet assay and micronucleus assay were carried out to evaluate genotoxicity. The results showed that, compared to the control group, QCT at the concentration ranges of 2–16?μg/mL significantly decreased L02 cell viability, which was significantly attenuated with curcumin pretreatment (2.5 and 5?μM). In addition, QCT significantly increased cell oxidative stress, characterized by increases of intracellular ROS level, while decreased endogenous antioxidant biomarkers GSH level and SOD activity (all p?<?0.05 or 0.01). Curcumin pretreatment significantly attenuated ROS formation, inhibited the decreases of SOD activity and GSH level. Furthermore, curcumin significantly reduced QCT-induced DNA fragments and micronuclei formation. These data suggest that curcumin could attenuate QCT-induced cytotoxicity and genotoxicity in L02 cells, which may be attributed to ROS scavenging and anti-oxidative ability of curcumin. Importantly, consumption of curcumin may be a plausible way to prevent quinoxaline 1,4-dioxides-mediated oxidative stress and genotoxicity in human or animals.     

Vinca alkaloids cause aberrant ROS-mediated JNK activation, Mcl-1 downregulation, DNA damage, mitochondrial dysfunction, and apoptosis in lung adenocarcinoma cells

Vinca alkaloids are clinically used to inhibit the growth of malignancy by interfering with microtubule polymerization. The purpose of this study was to identify the molecular mechanisms underlying growth inhibition as well as apoptosis in vinca alkaloid-treated lung adenocarcinoma cells. Consistent with nocodazole, treatment with vinorelbine (VNR) caused mitotic prometaphase arrest in a time-dependent manner, accompanied by cell apoptosis, dependent on both dose and time. VNR sequentially induced mitochondrial transmembrane potential (MTP) loss and caspase-dependent apoptosis following myeloid cell leukemia (Mcl) 1 downregulation. Prolonged activation of c-Jun N-terminal kinase (JNK) was required for vinca alkaloid- and nocodazole-induced apoptosis but not cell cycle arrest. Vinca alkaloids and nocodazole caused glutathione/reactive oxygen species (ROS) imbalance, and inhibiting ROS prevented prolonged JNK activation, decreased Mcl-1 levels, MTP loss, and apoptosis. Notably, cell size and granularity were enlarged in stimulated cells; unexpectedly, many ROS-producing mitochondria were accumulated followed by aberrant JNK-mediated mitochondrial dysfunction. Unlike cisplatin, which causes DNA damage in each phase of the cell cycle, VNR and nocodazole induced aberrant JNK-regulated DNA damage in prometaphase; however, inhibiting ATM (ataxia telangiectasia, mutated) and ATR (ATM and Rad3-related) did not reverse mitotic arrest or apoptosis. These results demonstrate an essential role of ROS in vinca alkaloid-induced aberrant JNK-mediated Mcl-1 downregulation and DNA damage followed by mitochondrial dysfunction-related apoptosis but not mitotic arrest.