红景天苷对人舌癌细胞的凋亡作用及机制研究

目的：探讨红景天苷对人舌癌细胞增殖和凋亡的影响,并解释其分子机制,为舌癌的临床治疗提供新的思路。方法：CCK-8法和克隆形成实验检测红景天苷药物对人舌癌细胞增殖能力的影响;Annexin V/PI双染法和TUNEL法检测细胞凋亡情况;PI染色法检测红景天苷对细胞周期的影响;细胞迁移法检测红景天苷对舌癌细胞迁移的影响;免疫印迹法和实时荧光定量PCR法检测细胞内凋亡相关蛋白Bax、Bcl-2和Caspase 3的表达水平;免疫印迹法检测细胞内ERK和PI3K/AKT信号通路关键蛋白的表达水平的变化。结果：红景天苷能显著抑制人舌癌细胞的增殖且呈剂量依赖性。Annexin V/PI双染法和TUNEL法显示,随着红景天苷浓度的增加,Tca-8113细胞凋亡明显增多。PI染色法显示,红景天苷处理细胞后,G2/M期细胞明显减少。细胞迁移实验表明,红景天苷作用于舌癌细胞后,随着给药浓度的逐渐升高,细胞的迁移能力明显降低。免疫印迹结果表明,红景天苷可以使Bax和Cleaved caspase 3蛋白表达水平升高,Bcl2的蛋白表达水平降低。同时,红景天苷可以抑制ERK和PI3K/AKT信号通路。结论：红景天苷可以明显抑制人舌癌Tca-8113细胞的增殖,诱导其凋亡,其作用机制可能与抑制ERK和PI3K/AKT信号通路有关。     

2-苯基-3,4-噻二唑[3,2-α]并嘧啶-7-酮对宫颈癌HeLa细胞增殖、凋亡及相关因子表达的影响

目的研究新化学合成药物2-苯基-3,4-噻二唑[3,2-α]并嘧啶-7-酮(TPh)对宫颈癌HeLa细胞增殖、凋亡及相关因子表达的作用。方法MTT法检测空白对照组和不同浓度TPh(0.6、0.4、0.2、0.1和0.05μmol·mL^-1)组培养24、48、72 h后对HeLa细胞增殖的作用,流式细胞术检测TPh对HeLa细胞周期的影响,Annexin-V FITC/PI双染检测TPh对HeLa细胞凋亡的影响,JC-1染色观察TPh对HeLa细胞线粒体膜电位的影响,Western blotting法检测凋亡相关因子表达情况。结果与空白对照组相比,TPh组Hela细胞增殖被显著抑制,HeLa细胞G0/G1期比例增加、G2/M期比例减少,HeLa细胞线粒体膜电位降低,HeLa细胞caspase-3、caspase-8、caspase-9表达增加、凋亡细胞比率增加(P<0.05,P<0.01),且呈剂量依赖性趋势。结论TPh可抑制宫颈癌HeLa细胞增殖,并促进凋亡,这可能与其激活细胞内源性和外源性凋亡途径有关。     

白屈菜红碱对人结直肠癌细胞凋亡的影响及其机制研究

目的：研究白屈菜红碱（Chelerythrine, CHE）对体外结直肠癌细胞的生长抑制作用,并探讨其作用机制。方法：MTT测定结肠癌细胞存活率,应用流式细胞仪检测CHE处理后ROS的积累和细胞凋亡情况,JC-1(5,5'',6,6''-四氯-1,1'',3,3''-四乙基苯并咪唑羰花青碘化物)荧光染料法检测细胞线粒体膜电位改变,应用荧光显微镜和Western blot法验证ROS积累诱导的线粒体功能障碍。结果：CHE对HCT-116和RKO细胞发挥剂量依赖性细胞毒作用,该作用与ROS介导的凋亡蛋白的表达有关。此外,CHE能够降低伴随线粒体功能障碍的线粒体膜电位。结论：CHE通过ROS介导的线粒体功能障碍和JNKs途径抑制CRC细胞生长并诱导细胞凋亡,提示CHE可能成为潜在的治疗CRC的候选药物。     

野马追总黄酮的体内外抗乳腺癌活性及机制研究

目的 探究野马追总黄酮(TFE)的体内外抗乳腺癌活性及相关的作用机制。方法 MTT法检测TFE对乳腺癌细胞(MCF-7、T47D、MDA-MB-231、MDA-MB-468)和人正常乳腺上皮细胞MCF-10A增殖的影响;流式细胞术检测TFE对乳腺癌细胞周期和凋亡的影响;吖啶橙(AO)染色法检测TFE对细胞自噬的影响;Western blot检测细胞周期、凋亡、自噬相关蛋白的变化,以及PI3K/Akt信号通路相关蛋白的变化;建立MDA-MB-231细胞的裸鼠移植瘤模型,观察TFE的体内抗肿瘤活性。结果 TFE能明显抑制乳腺癌细胞增殖,并呈剂量依赖性,而对人正常乳腺上皮细胞无显著的抑制作用;TFE能增加G₂/M期细胞比例,上调p-cdc-2蛋白表达,下调cdc-2和Cyclin B1蛋白表达;TFE能显著增加乳腺癌细胞凋亡比例,上调促凋亡蛋白Bad和Bax表达,下调抗凋亡蛋白Bcl-2表达;TFV能够诱导乳腺癌细胞产生自噬,自噬相关蛋白LC3-II表达上升,p62表达降低;TFE能够抑制显著抑制PI3K和Akt的磷酸化水平;TFE能够在体内抑制裸鼠肿瘤的体积。结论 TFE通过抑制PI3K/Akt信号通路,阻滞细胞周期、诱导凋亡和自噬,进而在体内外抑制乳腺癌细胞的生长。     

Shenmai Injection attenuated doxorubicin-induced cardiac dysfunction via maintaining mitochondrial homeostasis

OBJECTIVE Shenmai Injection(SMI) is widely used in the treatment of cardiovascular diseases, such as heart failure and myocardial ischemia. In clinic, SMI showed protective effects on doxorubicin(Dox)-induced cardiac toxicity. In current study, we investigate the mitochondrial protective mechanisms of SMI on Dox-induced myocardial injury.METHODS C57 BL/6 mice were divided into four groups:(1) control group;(2) Dox injury group;(3) SMI+Dox group and dexrazoxane(DRZ)+Dox group. Dex was a positive control. Myocardial injury was evaluated by echocardiography, HE and TUNEL staining, myocardial markers measurement. H9 C2 cardiomyocytes pretreatment with SMI for 24 h were exposed to Dox. Cell viability and apoptosis were measured by CCK8, Hoechst33342 staining, and Annexin V/PI staining.Mito SOX, mitochondrial membrane potential, and mitochondrial respiratory function were measured to evaluate mitochondrial function. RESULTS SMI decreased mortality rate of Dox-injected mice, serum CK and CK-MB levels in vivo.SMI significantly prevented Dox-induced cardiac dysfunction and apoptosis and increased expression level of PI3 K,p-Akt, and p-GSK-3β. Moreover, SMI significantly inhibited Dox-induced apoptosis, mitochondrial ROS production, and reduction of mitochondrial membrane potential in H9 C2 cells. Mechanismly, the cardio-protective effect of SMI was suppressed by PI3 K inhibitor LY294002. CONCLUSION SMI prevents Dox-induced cardiotoxicity and mitochondrial damage through activation of PI3 K/Akt signaling pathway.     

Shenmai injection attenuated doxorubicin-induced cardiac dysfunction via maintaining mitochondrial homeostasis

OBJECTIVE Shenmai injection(SMI) is widely used in the treatment of cardiovascular diseases,such as heart failure and myocardial ischemia. In clinic,SMI showed protective effects on doxorubicin(Dox)-induced cardiac toxicity. In current study, we investigate the mitochondrial protective mechanisms of SMI on Doxinduced myocardial injury. METHODS C57 BL/6 mice were divided into four groups:(1) control group;(2) Dox injury group;(3) SMI+Dox group;(4) dexrazoxane(DRZ)+Dox group. Dex as a positive control. Myocardial injury was evaluated by echocardiography, H&E and TUNEL staining, myocardial markers measurement. H9C2 cardiomyocytes pretreatment with SMI for 24 h were exposed to Dox. Cell viability and apoptosis were measured by CCK8, Hoechst33342 staining, and Annexin Ⅴ/PI staining.Mito SOX, mitochondrial membrane potential, and mitochondrial respiratory function were measured to evaluate mitochondrial function. RESULTS SMI decreased mortality rate of Dox-injected mice, serum CK and CK-MB levels in vivo. SMI significantly prevented Dox-induced cardiac dysfunction, apoptosis and mice survival, as well as increased expression level of PI3K, p-Akt, and p-GSK-3β Moreover, SMI significantly inhibited Dox-induced apoptosis,mitochondrial ROS production, reduction of mitochondrial membrane potential, and regulate mitochondrial respiration in H9C2 cells. Mechanismly, the cardio-protective effect of SMI was suppressed by PI3K inhibitor LY294002.CONCLUSION SMI prevents Dox-induced cardiotoxicity and mitochondrial damage through activation of PI3 K/Akt signaling pathway.     

Curcumin inhibits cervical cancer cell proliferation,migration and invasion by suppressing the PI3K/AKT signaling pathway via the miR-29b/KDM2A axis

In this study, we aimed to examine whether curcumin exerted its anti-tumor effects by regulating miR-29b/KDM2A in cervical cancer cells. The cell viability, migration and invasion were estimated in HeLa cervical cancer cells treated with curcumin. The effects of microRNA-29b (miR-29b) on biological behaviors of HeLa SiHa cells were also assessed. Potential target genes of miR-29b were predicted and confirmed using a luciferase reporter assay, and the effects of curcumin and miR-29b on the PI3K/AKTsignaling pathway were analyzed. Curcumin treatment inhibited cell proliferation, migration and invasion of HeLa cells (P<0.05). The miR-29b expression was promoted by curcumin treatment in HeLa cells (P<0.01), and miR-29b depletion could restore the effects of curcumin on cell proliferation, migration and invasion of HeLa cells (P<0.05). KDM2A was proved as a direct target gene of miR-29b, and the activity of the PI3K/AKT signaling could be regulated by curcumin and miR-29b (P<0.05). All the data revealed that curcumin played a protective role in cervical cancer. The proliferation, migration and invasion of cervical cancer cells were inhibited by curcumin through the miR-29b/KDM2A/PI3K/AKT pathway.     

西多福韦对HeLa细胞增殖抑制作用的研究

目的 研究抗病毒药西多福韦(CDV)对HeLa细胞的增殖抑制作用.方法 采用MTT法分析CDV和阳性对照药顺铂(DDP)对HeLa细胞增殖活力的影响;利用细胞集落形成实验和Giemsa染色技术,观察CDV和DDP导致HeLa细胞凋亡情况;利用流式细胞仪观察CDV和DDP对细胞凋亡及细胞周期影响.结果 MTT法和细胞集落实验结果显示CDV能明显抑制HeLa细胞的增殖活性;流式细胞仪测定显示CDV和DDP使HeLa细胞阻滞于细胞周期的S期,并诱导HeLa细胞发生凋亡.结论 CDV对HeLa细胞具有明显的增殖抑制作用,并能诱导细胞凋亡,CDV有可能成为宫颈癌治疗的另一策略.     

木犀草素对K562细胞增殖与凋亡的影响及其机制

目的　探讨木犀草素对K562细胞增殖、凋亡的影响及其作用机制。方法　取对数生长期K562细胞分别加入0、10、25、50、100 μmol/L木犀草素培养24 h、48 h、72 h,采用CCK-8法检测细胞增殖抑制率;K562细胞分别加入0、25、50 μmol/L木犀草素培养48 h,采用流式细胞术检测细胞凋亡情况;K562细胞分别加入0、10、50、100 μmol/L木犀草素培养48 h,采用Westem blot检测B细胞淋巴瘤2蛋白（Bcl-2）、Bcl-2相关X蛋白（Bax）、多聚ADP核糖聚合酶（PARP）、Cleaved-PARP、半胱氨酸天冬氨酸蛋白水解酶3（Caspase3）、Cleaved-Caspase3、蛋白激酶B（AKT）、磷酸化AKT（p-AKT）、断裂点簇集区蛋白（BCR）、c-abl癌基因1（c-ABL）BCR-ABL蛋白表达。结果　CCK-8检测结果显示,随木犀草素浓度增加及作用时间的延长,K562细胞增殖抑制率均呈增长趋势（P＜0.05）。流式细胞仪检测结果显示,木犀草素0、25、50 μmol/L组K562细胞的凋亡率依次升高（分别为8.21%±0.55%、23.43%±1.50%和40.47%±2.97%）。Western blot结果显示,Bax、Cleaved-PARP、Cleaved-Caspase3表达水平随木犀草素浓度的增加而升高（P＜0.05）。木犀草素0、10、50 μmol/L组PARP蛋白表达水平依次升高（P＜0.05）,100 μmol/L组与50 μmol/L组差异无统计学意义。100 μmol/L组Caspase3、Bcl-2蛋白表达水平均低于其余组（P＜0.05）。50、100 μmol/L组p-AKT蛋白表达水平低于0、10 μmol/L组,100 μmol/L组低于50 μmol/L组。50 μmol/L组BCR-ABL融合蛋白表达水平高于0 μmol/L组,100 μmol/L组低于0、50 μmol/L组（P＜0.05）。结论　木犀草素可抑制K562细胞增殖,促进细胞凋亡,其机制可能与调控BCR-ABL蛋白表达及PI3K/AKT信号通路有关。     

Inhibition of cathepsin S induces autophagy and apoptosis in human glioblastoma cell lines through ROS-mediated PI3K/AKT/mTOR/p70S6K and JNK signaling pathways

Cathepsin S is a lysosomal cysteine protease that is overexpressed in various cancer models and plays important role in tumorigenesis, however the mechanisms are unclear. In the present study, we found that inhibition of cathepsin S induced autophagy and mitochondrial apoptosis in human glioblastoma cells. Blockade of autophagy by either a chemical inhibitor or RNA interference attenuated cathespin S inhibition-induced apoptosis. Furthermore, autophagy and apoptosis induction was dependent on the suppression of phosphatidylinositide 3-kinases/protein kinase B/mammalian target of rapamycin/p70S6 kinase (PI3K/AKT/mTOR/p70S6K) signaling pathway and activation of c-Jun N-terminal kinase (JNK) signaling pathway. In addition, reactive oxygen species (ROS) served as an upstream of PI3K/AKT/mTOR/p70S6K and JNK signaling pathways. In conclusion, the current study revealed that cathepsin S played an important role in the regulation of autophagy and apoptosis in human glioblastoma cells.     

Augmentation of oridonin-induced apoptosis observed with reduced autophagy

Our previous studies showed that oridonin could induce apoptosis in HeLa cells; and in this study, we further investigated autophagy induced by oridonin in HeLa cells and the relationship between apoptosis and autophagy. HeLa cells were exposed with oridonin after 3-methyladenine (3-MA) pre-culture, and we evaluated the growth inhibitory ratio, morphologic changes, DNA fragmentation, proteins expression as well as autophagic and apoptotic levels. Oridonin inhibited the proliferation of HeLa cells in vitro and induced autophagy. Oligonucleosomal fragementation of DNA as well as increased activities of Bax proteins were induced by oridonin, but the expression of p-Bcl-2 protein was reduced. In the condition of oridonin-treatment, when the inhibitor of phosphoinositide 3-kinase (PI3K), wortmannin, was applied, the autophagic level was significantly decreased, while the apoptotic level was increased, indicating that PI3K is a key regulator of both autophagy and apoptosis. Akt, down-stream factor of PI3K, was activated in autophagic process but suppressed in apoptosis in this study. In addition, when autophagy was blocked by 3-MA, the expression of SIRT-1 was decreased, indicating SIRT-1 contributed to autophagy. Taken together, oridonin simultaneously induced HeLa cell both apoptosis and autophagy in HeLa cells, and inhibition of autophagy contributes to upregulation of apoptosis.     

黄芪中有效成分对人食道癌HCE-4细胞凋亡的影响

目的 研究黄芪有效成分芒柄花素与黄芪甲苷IV对人食道癌HCE-4细胞的凋亡作用及其分子机制。方法 MTT法检测黄芪提取物(阳性对照药)、芒柄花素与黄芪甲苷IV对人食道癌HCE-4细胞的增殖抑制作用,Annexin V-FITC/PI双染法检测3种药物诱导HCE-4细胞凋亡的能力,Western blotting法检测凋亡相关蛋白的表达变化。结果 芒柄花素与黄芪甲苷IV对HCE-4细胞具有良好的生长抑制作用,其效果呈浓度依赖性;芒柄花素与黄芪甲苷IV能够诱导HCE-4细胞的凋亡且呈时间依赖性;芒柄花素与黄芪甲苷IV处理HCE-4细胞后,抗凋亡蛋白p-AKT的表达量明显减少,促凋亡蛋白cleaved-caspase-3表达量增加。结论 黄芪有效成分芒柄花素与黄芪甲苷IV通过调控AKT信号转导途径诱导人食道癌HCE-4细胞凋亡。     

Colchicine-induced apoptosis in human normal liver L-02 cells by mitochondrial mediated pathways

Colchicine is an alkaloid that has been widely used to treat gout. It also has a curative effect on cancer. Although many studies have shown that its effect on cell apoptosis was mediated by the activation of caspase-3, the pathways involved in the process remained obscure. Here we show some evidence regarding the missing information using human normal liver cells L-02 in our study. The effect of colchicine on apoptosis in L-02 cells and the apoptosis-associated signaling pathways were determined using different tests including cell viability assay, Annexin V and propidium idodide binding, PI staining, Hoechst 33342 staining, mitochondrial membrane potential assay, caspase activity assay and Western blot analysis. We found that colchicine-induced a dose-dependent drop of cell viability in L-02 cells; early apoptosis happened when cells were treated with 0.1 μM of colchicine. The colchicine-induced loss of mitochondrial membrane potential, activation of caspase-3 and 9, up-regulation of Bax and down-regulation of Bcl-2 showed an evidence for the colchicine activity on apoptosis, at least, by acting via the intrinsic apoptotic pathway.     

藤黄酸抑制人卵巢透明细胞癌ES-2细胞增殖和促进其凋亡的机制研究

目的 研究藤黄酸对人卵巢透明细胞癌ES-2细胞增殖和凋亡的影响及其潜在分子机制。方法 采用CCK-8检测藤黄酸对不同类型人卵巢癌细胞增殖的影响;克隆形成实验检测藤黄酸对ES-2细胞克隆形成的影响;细胞划痕愈合实验及Transwell实验分别检测藤黄酸对ES-2细胞迁移与侵袭的影响;Caspase-Glo 3/7试剂盒检测藤黄酸对ES-2细胞内Caspase 3/7活性的影响;Annexin V-FITC/PI双染色法分析藤黄酸对ES-2细胞凋亡的影响;Western blotting检测不同浓度藤黄酸对ES-2细胞中Bcl-2、Bax、PTEN、PI3K、p-AKT、p-mTOR水平的影响。结果 人卵巢癌ES-2、HO8910、SKOV3、OVCAR3细胞中,藤黄酸对卵巢透明细胞癌ES-2细胞的增殖抑制作用最明显,且随着藤黄酸浓度和作用时间的增加而增强;藤黄酸可抑制ES-2细胞的克隆形成、迁移及侵袭、增强Caspase 3/7活性并促进细胞凋亡,且呈浓度依赖性;藤黄酸可下调ES-2细胞中Bcl-2、PI3K、p-AKT及p-mTOR的表达,上调Bax、PTEN的表达,且呈浓度依赖性。结论 藤黄酸能够抑制卵巢透明细胞癌ES-2细胞的增殖、迁移和侵袭,并诱导其凋亡,其作用机制可能与调控PI3K/AKT/mTOR信号通路相关。     

Cytotoxic effect and apoptotic mechanism of tanshinone A, a novel tanshinone derivative, on human erythroleukemic K562 cells

Tanshinone A is a novel derivative of phenanthrene-quinone extracted from Salvia miltiorrhiza BUNGE, a traditional herbal medicine. Cytotoxic effect of tanshinone A was observed in this study. Additionally its mechanism of promoting apoptosis was also investigated. MTT and SRB assays were applied to measure the effects of tanshinone A on the cell viability, the cell cycle distribution and cell apoptosis were measured by flow cytometry using PI staining and Annexin V/PI double staining method respectively. The changes of mitochondrial membrane potential were also detected by flow cytometry. Spectrophotometric method was used to detect the changes of caspase-3 activity. Western blotting assay was used to evaluate the expression of bcl-2, bax and c-Myc proteins. Results indicated that tanshinone A displayed a significant inhibitory effect on the growth of K562 cells in a dose- and time-dependent manner, and showed obvious minor damage to LO2 cells. Tanshinone A could arrest K562 cells in the G(0)/G(1) phase and induce apoptosis, decrease the mitochondrial transmembrane potential, decrease the expressions of bcl-2 and c-Myc proteins, increase the expression of bax protein and the activity of caspase-3. Accordingly, it was presumed that the apoptosis induction may be through the endogenous pathway. Subsequently, tanshinone A could be a promising candidate in the development of a novel antitumor agent.     

双氢青蒿素诱导人肿瘤细胞凋亡及其分子机制的研究

目的:研究双氢青蒿素诱导人白血病细胞凋亡作用及其机制。方法:用双氢青蒿素处理K562细胞,通过MTT比色法检测细胞增殖抑制的效果;荧光显微镜观察细胞的凋亡;逆转录-聚合酶链反应(RT-PCR)检测半胱氨酰-天冬氨酸蛋白酶-3(caspase-3)的表达,蛋白印迹技术(Western blot)检测线粒体、胞浆细胞色素C的表达。结果:双氢青蒿素处理K562细胞48h后MTT检测,570nm波长处测定半数细胞抑制浓度为8×10-5mol.L-1;Hoechst33342/PI双荧光染色可观察到明显核固缩、凝集等细胞增殖抑制表现;RT-PCR检测到caspase-3的表达,Western-blot检测线粒体细胞色素C表达下调,胞浆中细胞色素C表达阳性。结论:双氢青蒿素能够在体外抑制人白血病细胞增殖并诱导其凋亡,而促进细胞色素C的释放,激活caspase-3可能是其作用机制之一。     

青藤碱通过调控NF-kB信号通路抑制胰腺癌Capan-1细胞增殖并诱导细胞凋亡

目的 探讨青藤碱(Sinomenine, SIN)对胰腺癌Capan-1细胞增殖及凋亡的影响。方法 CCK8法检测细胞活力,光学显微镜下观察细胞凋亡形态,DAPI/TUNEL双染检测细胞凋亡,流式细胞术Annexin V-FITC/PI法检测细胞凋亡率,Western blot检测裂解型Caspase-3蛋白(cleaved caspase-3)、细胞核内NF-κB(Nuclear factor-kappa-binding)蛋白表达。结果 CCK8结果表明青藤碱抑制胰腺癌Capan-1细胞增殖,并具有时间浓度依赖性;光学显微镜观察结果表明,在青藤碱作用下,Capan-1细胞皱缩、变圆、脱落增多;DAPI/TUNEL染色结果表明,在青藤碱作用下,Capan-1细胞凋亡增多;流式细胞术Annexin V-FITC/PI结果表明,在青藤碱作用下,Capan-1细胞凋亡率增高;Western blot结果表明,在青藤碱作用下,Capan-1细胞cleaved caspase-3、核内NF-κB表达降低,NF-κB信号通路激活剂TNF-α逆转青藤碱对Capan-1细胞增殖的抑制作用、凋亡率的升高作用和cleaved caspase-3表达的下调作用。结论 青藤碱通过调控NF-κB信号通路抑制胰腺癌Capan-1细胞增殖并诱导细胞凋亡。     

雷公藤红素通过PI3K/AKT/mTOR信号通路对胃癌MFC细胞增殖和凋亡的影响

目的研究雷公藤红素对胃癌MFC细胞内PI3K/AKT/mTOR信号通路的影响及其对胃癌MFC细胞增殖的抑制和促凋亡作用机制。方法分别设置对照组和雷公藤红素低、中、高剂量(5、10、20 mmol/L)组,采用MTT法检测雷公藤红素对胃癌MFC细胞增殖的影响;采用流式细胞技术检测雷公藤红素对胃癌MFC细胞周期的影响及MFC细胞凋亡的情况;Western blotting检测雷公藤红素对胃癌MFC细胞内凋亡相关蛋白Bax、Bcl-2和PI3K、AKT和mTOR蛋白表达的影响;实时定量PCR检测雷公藤红素对胃癌MFC细胞内PI3K、AKT和mTORm RNA表达的影响。结果与对照组比较,雷公藤红素能够呈剂量相关性地显著抑制胃癌MFC细胞的增殖(P0.05);与对照组比较,雷公藤红素能够呈剂量相关性地显著引起MFC细胞G2/M期阻滞(P0.05);与对照组比较,雷公藤红素能够呈剂量相关性地显著促进MFC细胞的凋亡(P0.05);与对照组比较,雷公藤红素能够呈剂量相关性地显著抑制MFC细胞内抗凋亡蛋白Bcl-2蛋白和PI3K、AKT、mTOR蛋白的表达(P0.05),显著促进MFC细胞内促凋亡蛋白Bax的表达(P0.05);与对照组比较,雷公藤红素能够呈剂量相关性地显著抑制MFC细胞内PI3K、AKT和mTORm RNA的表达(P0.05)。结论雷公藤红素能够抑制胃癌MFC细胞的增殖并促进胃癌MFC细胞凋亡,其作用机制可能是通过抑制胃癌MFC细胞内PI3K/AKT/mTOR信号通路中PI3K、AKT、mTOR等蛋白的表达而抑制胃癌MFC细胞的增殖,进而促进胃癌MFC细胞内促凋亡蛋白Bax的表达,同时抑制抗凋亡蛋白Bcl-2的表达,从而促进胃癌MFC细胞凋亡。     

金粉蕨素体外诱导人宫颈癌HeLa 细胞凋亡的机制

目的：探讨金粉蕨素（ONY）体外诱导人宫颈癌HeLa细胞凋亡的作用及其分子机制。方法体外培养人宫颈癌HeLa、CaSki、SiHa、ME180细胞,采用MTT法检测ONY对人宫颈癌HeLa、CaSki、SiHa、ME180细胞生长抑制率的影响;Hoechst33258染色检测ONY对人宫颈癌HeLa细胞凋亡的形态学变化;流式细胞术（FCM）检测ONY对HeLa细胞凋亡率的影响;DNA琼脂糖电泳检测HeLa细胞凋亡的DNA条带;Western blot分析凋亡相关蛋白表达变化。结果 ONY对人宫颈癌HeLa、CaSki、SiHa、ME180细胞生长有较强的抑制作用,呈剂量和时间依赖性,以HeLa细胞对ONY最为敏感,作用24 h的IC50值为10.48μg·mL-1。经ONY作用于HeLa细胞24 h后,细胞出现典型的凋亡形态学改变,表现出典型的凋亡特征的亚二倍体峰,且呈剂量依赖性,同时出现典型的凋亡DNA条带;同时,cytochrome c、caspase-9和caspase-3蛋白表达增加,bcl-2蛋白表达下调,而bax蛋白表达不变, Bcl-2/Bax比值下调（P〈0.05）。结论 ONY可能通过调控线粒体途径相关蛋白抑制宫颈癌HeLa细胞增殖并诱导其凋亡。