酸敏脂质体的制备及其生物学活性

目的　对不同方法制备酸敏脂质体进行包封率测定及形态学观察 ,探讨酸敏脂质体包裹反义寡核苷酸的生物学效应。方法　测定四种不同方法制备包裹1 2 5I IL 8脂质体的包封率 ;在超高倍显微分析仪下观察形态结构。用包裹细胞磷脂酶A2 (cPLA2 )反义寡核苷酸的酸敏脂质体转染U937细胞 ,提取细胞RNA ,采用反转录 聚合酶链反应 (RT PCR) ;免疫印迹 (Westernblot)法检测cPLA2 蛋白的表达。结果　反相蒸发法与钙融合法联合制备的酸敏脂质体具有良好地包封率和形态结构 ,经酸敏脂质体包裹cPLA2 反义寡脱氧核苷酸可明显抑制cPLA2基因及蛋白的表达。结论　制备所得的酸敏脂质体可良好的发挥其生物学效应     

脂质体包裹的反义寡核苷酸逆转耐甲氧西林金黄色葡萄球菌耐药性的研究

目的：用人工合成磷脂二棕榈酰磷脂酰胆碱（dipalmitoyl phosphatidylcholine，DPPC），二肉豆蔻酰磷脂酰甘油（dimyristoyl phosphatidylglycerol，DMPG）制备反义寡核苷酸阴离子脂质体并研究脂质体包裹的抑制耐甲氧西林金黄色葡萄球菌（methicillin resistant staphylococcus aureus，MRSA）耐药基因表达信号传导通路中BlaRlmRNA表达的反义寡核苷酸（antisense phosphothioate oligodeoxynucleotides，AS-ODNs）对MRSA耐药性的影响。方法：设计合成AS-ODNs；薄膜分散冻干法制备其脂质体；透射电镜观察脂质体的形态；离心纯化脂质体并用紫外分光光度计测定包封率、渗漏率；振荡法检测体外释放度；平板克隆形成实验计数菌落数CFU；微量法测定细菌生长曲线。结果：反义寡核苷酸阴离子脂质体大小均匀，为圆球体，包封率为77．38％，冷冻条件下保存1月后渗漏率为0．18％，体外释放度实验表明24h后约60％的药物从脂质体中释放，反义寡核苷酸脂质体可显著抑制MRSA生长，脂质体包裹的不同剂量的AS-ODNs中MRSA的菌落形成单位（CFU）与空白对照组比较明显减少，具有剂量依赖性，且效果明显优于未被脂质体包裹的AS-ODNs。结论：采用薄膜分散冻干法制备反义寡核苷酸阴离子脂质体，包封率较高，质量稳定，反义寡核苷酸脂质体能逆转MRSA的耐药性，效果明显优于单用AS-ODNs，可作为反义寡核苷酸进入细菌的载体。     

mdr1反义寡核苷酸增强多药耐药肝癌细胞HepG2/ADM化疗敏感性的实验研究

目的：探讨在体外实验条件下mdr1反义寡核苷酸对多药耐药肝癌细胞株化疗敏感性的影响。方法以肝癌细胞HepG2/ADM为研究对象,设立mdr1反义寡核苷酸组和空白试剂组作对照,利用脂质体包载肿瘤耐药基因mdr1的反义寡核苷酸进行细胞转染,通过反转录聚合酶链反应（RT-PCR）、免疫印迹实验（Western blotting）分别检测mdr1基因mRNA和P-gp蛋白表达,通过MTT实验检测细胞转染前后对阿霉素（ADM）、顺铂（DDP）和5-氟尿嘧啶（5-FU）的化疗敏感性。结果 HepG2/AMD肝癌细胞经反义寡核苷酸处理后,mdr1 mRNA、P-gp蛋白表达水平均明显降低,对ADM、DDP 和5-FU的化疗敏感性明显增强。结论反义寡核苷酸能在体外有效增加肝癌细胞HepG2/ADM对化疗药物的敏感性。     

谷甾醇葡萄糖苷和卞泽双重修饰肝实质细胞靶向阳性脂质体介导的基因转染和抗乙肝病毒作用

目的研究载乙型肝炎病毒(HBV)反义寡核苷酸的双重表面修饰肝实质细胞靶向阳性脂质体的基因转染，抗乙肝病毒作用和其介导基因转染的机制。方法以3β-［N-(N′,N′-二甲氨基乙基)-氨甲酰基］胆固醇(DC-Chol)和二棕榈酰磷脂酰胆碱(DPPC)为脂材，分别以谷甾醇葡萄糖苷(sito-G)和卞泽(Brij 35)为膜表面修饰成分，制备载HBV反义寡核苷酸的阳性脂质体。采用大鼠原代肝实质细胞和人肝癌细胞HepG 2.2.15，通过流式细胞分析、荧光显微镜观察和酶联免疫吸附试验(ELISA)，考察脂质体对基因转染的促进作用及其病毒抑制作用；通过评价渥曼青霉素、尼日利亚菌素以及无涎胎球蛋白对其病毒抑制作用的影响，探讨其转染机制。结果以sito-G和Brij 35对脂质体进行双重表面修饰，显著提高了脂质体的转染率和病毒抑制作用；荧光显微镜下观察到较强转染，反义寡核苷酸的胞内分布以在细胞核中为主；渥曼青霉素、尼日利亚菌素和无涎胎球蛋白均不同程度地降低了载反义寡核苷酸脂质体的病毒抑制作用。结论Brij 35和sito-G双重修饰阳性脂质体显示出较高的基因转染效率和显著的病毒抑制作用，其基因转染过程以内吞和膜融合为主，并表现出肝实质细胞表面去唾液酸糖蛋白受体 (ASGPR)的靶向选择性。     

透明质酸磷脂酰衍生物修饰姜黄素脂质体的制备及细胞毒性

采用逆相蒸发法制备了姜黄素脂质体.由于透明质酸脂溶性较差,先将其制成磷脂酰衍生物(透明质酸-磷脂酰乙醇胺,HA-DOP E),再用以修饰载药脂质体.所得脂质体的平均包封率为89％,该衍生物与脂质体的结合率为72％.脂质体于4℃保存30d包封率无明显改变,但结合率有轻微下降.细胞毒性试验显示,游离姜黄素、姜黄素脂质体及修饰后脂质体对高表达CD44受体的人肺腺癌A549细胞的IC50值分别为0.054、0.032和0.021 μmol/ml,未修饰和修饰后的姜黄素脂质体对低表达CD44受体的人肝癌HepG2细胞作用相当.     

甘草次酸修饰多西紫杉醇脂质体的制备和体外抑瘤效果

目的:制备甘草次酸(GA)修饰的多西紫杉醇脂质体,并初步考察其体外抗肿瘤效果.方法:化学合成甘珀酸十八醇酯(18-GA-Suc)作为修饰材料,采用薄膜分散法制备甘草次酸修饰的多西紫杉醇脂质体,考察影响脂质体包封率的因素.采用MTT法评价脂质体对HepG2细胞的体外抑瘤效果.结果:18-GA-Suc修饰的DX脂质体的体外抑瘤效果强于未修饰的DX脂质体,并且抑瘤效果随着载体中18-GA-Suc的增加而增强.结论:甘草次酸修饰的脂质体有望成为新型肝靶向的抗肿瘤载体.     

血管内皮生长因子反义寡核苷核对前列腺癌细胞PC3生长特性的影响

目的探讨血管内皮生长因子(VEGF)反义寡核苷核对前列腺癌细胞PC3生长特性的影响。方法采用新型脂质体Oligofectamine携带VEGF反义寡核苷酸转染激素非依赖性前列腺癌细胞PC3,实验分为对照组、反义寡核苷核苷酸组和正义寡核苷酸组。Western Blot杂交的方法检测细胞VEGF蛋白的表达,四甲基偶氮唑蓝法(MTT)检测细胞增殖变化,流式细胞仪检测细胞凋亡情况。结果新型脂质体可以携带VEGF反义寡核苷酸转染前列腺癌细胞PC3,与对照组和正义寡核苷酸组比较,反义寡核苷酸组细胞VEGF蛋白的表达明显下降,增殖受到明显抑制,凋亡率增加。结论新型脂质体Oligofectamine可以携带VEGF反义寡核苷酸成功转染前列腺癌细胞PC3,抑制VEGF的表达,进而抑制肿瘤细胞的增殖,促进其凋亡。     

阳离子脂质材料和聚乙二醇对脂质体细胞转染率及膜流动性的影响阳离子脂质材料和聚乙二醇对脂质体细胞转染率及膜流动性的影响

目的制备包封荧光素钠（FS）的脂质体，考察阳离子脂质材料（DC-chol）和聚乙二醇（PEG）对脂质体包封率、细胞转染率及膜流动性的影响。方法以FS作为模型物质，制备并分离脂质体，测定脂质体包封率；通过观察荧光光谱的变化考察FS与脂质体膜之间的相互作用；以HepG₂ 2.2.15为细胞模型观察脂质体对FS细胞转染率的影响；通过荧光偏振技术考察阳离子脂质材料和PEG对脂质体膜流动性的影响。结果阳离子脂质材料和PEG能提高脂质体包封率（0.64％～86.57％）、细胞转染率（2.18％～48.46％）及脂质体膜流动性，PEG分子质量的增大有利于包封率、转染率的提高，并增加脂质体膜的流动性。结论在脂质体处方中加入阳离子脂质材料和高分子量的PEG有利于提高包封率、细胞转染率及增加脂质体膜的流动性。     

反义白介素-1受体相关激酶-1寡核苷酸抑制白介素-1诱导的NF-κB活化

目的 :研究反义白介素 1受体相关激酶 1(IRAK 1)寡核苷酸对核因子 κB(NF κB)活化的影响。方法 :Lipofectin介导反义IRAK 1寡核苷酸转染HepG2细胞 ,以逆转录PCR法检测IRAK 1mRNA表达水平 ,用SandwichELISA法检测NF κB的活化。结果 :反义IRAK 1寡核苷酸抑制IRAK 1mRNA表达 ;反义IRAK 1寡核苷酸呈剂量 (1～ 8μg)和时间 (5～ 2 4h)依赖性地抑制NF κB活化 ,反义IRAK 1寡核苷酸 4μg与细胞共孵育 8h时抑制效果最好。结论 :反义IRAK 1寡核苷酸通过阻断IRAK 1表达抑制NF κB活化 ,预示反义IRAK 1寡核苷酸可潜在抑制IL 1的炎症活性。     

载基因纳米阳离子脂质体前体制剂的制备及性质研究

目的:制备稳定的载反义寡核苷酸的阳离子脂质体前体制剂。方法:以磷脂-二油酰磷脂酰乙醇胺(dioleoylphophatidylethanolamine,DOPE)-十八胺-胆固醇为类脂成分,采用薄膜超声-挤压制备空白阳离子脂质体,吸附-冷冻干燥法制备载反义寡核苷酸阳离子脂质体前体。激光粒度仪测定冷冻干燥前后脂质体Zeta电位及粒径,透射电镜观察其形态,葡聚糖凝胶柱分离未包封的反义寡核苷酸,紫外法测定冻干前后的载药率。结果:海藻糖与甘露醇及甘氨酸为较好的冻干保护剂,制得的阳离子脂质体前体带正电荷,规则球形,大小较均匀,海藻糖作为保护剂复溶前后平均粒径为175和320 nm左右,复融前后Zeta电位值在+32和+40 mV左右,脂质体的载药率复溶前后分别为87.6%与83.21%。结论:海藻糖作为冻干保护剂,薄膜超声挤压法与冷冻干燥法结合,可成功制备反义寡核苷酸阳离子脂质体前体制剂,稳定性大大改善。     

Delivery of different length poly(L-lysine)-conjugated ODN to HepG2 cells using N-stearyllactobionamide-modified liposomes and their enhanced cellular biological effects

Short (14-20-mer range) synthetic oligodeoxynucleotides (ODNs) allow specific modulation of cellular gene expression at various stages, thus providing a versatile tool for fundamental studies and a rational approach to anticancer chemotherapy. However, several problems, such as metabolic stability, efficient cell internalization of ODNs and their efficient entrapment into liposomes continue to markedly limit this approach. To improve the target specificity and biological activity of ODN, three different length of poly(L-lysine) (PLL) were conjugated to ODN and these conjugates were encapsulated in N-stearyllactobionamide (N-SLBA)-modified liposomes, N-SLBA is a ligand for the asialoglycoprotein receptor. Then, we investigated their effects on cell cycle and survivin protein levels of HepG2 cells. The results showed that the encapsulation efficiency was improved because the polycationic charges of PLL neutralized the polyanionic charges of ODN. Among them, PLL (M(W) 2000 and 10,000)-conjugated ODN encapsulated in N-SLBA liposomes induced apoptosis of HepG2 cells and highly inhibited survivin gene expression.     

Evaluation of Generation 2 and 3 Poly(Propylenimine) Dendrimers for the Potential Cellular Delivery of Antisense Oligonucleotides Targeting the Epidermal Growth Factor Receptor

PURPOSE: To evaluate low generation, G2 and G3, poly(propylenimine) dendrimers for the potential cellular delivery of antisense oligonucleotides (ODNs) targeting the epidermal growth factor receptor (EGFR) in A431 epidermoid carcinoma cells. METHODS: Cell cytotoxicity of the dendrimers was evaluated using trypan blue exclusion assays. Cellular uptake studies of fluorescently labeled ODNs were performed using fluorescence-activated cell sorting analysis. Intracellular fate of dendrimer-delivered ODNs was assessed in both fixed and live cells using fluorescent microscopy. Antisense ODN activity was assessed in terms of cancer cell growth, inhibition of target EGFR protein, and reduction in mRNA levels. RESULTS: G2 dendrimer (DAB-8) was less toxic than G3 (DAB-16) dendrimer in A431 cells, with IC50 of >175 and approximately 30 microg/ml, respectively. Uptake of fluorescently labeled ODN:dendrimer complexes was increased by up to 100-fold compared to a marker of fluid-phase endocytosis and up to 9-fold over free ODN at the optimal dendrimer:ODN (w/w) ratio of 5:1. Uptake of dendrimer:ODN complexes was significantly reduced at 4 degrees C (p < 0.05). Live cell fluorescent microscopy resulted in an intracellular distribution of dendrimer:ODN complexes that was suggestive of endocytic uptake; in contrast, cell fixation resulted in an artefactual nuclear localization. Treatment of A431 cells with anti-EGFR antisense ODN:dendrimer complexes inhibited cell growth, protein, and mRNA expression to levels comparable to Oligofectamine-mediated delivery. CONCLUSIONS: G2 and G3 poly(propylenimine) dendrimers markedly improved the delivery and activity of ODNs and thus may represent general reagents for the delivery of ODNs to cells in culture.     

吡非尼酮对肝癌HepG2细胞增殖和凋亡的影响(“豪森杯”优秀论文三等奖)

目的：研究吡非尼酮（pirfenidone,PF）对人肝癌细胞系HepG2增殖和凋亡的影响。方法：CCK-8法测定不同浓度PF对HepG2细胞增殖活性的影响;Hoechst 33258荧光染色法观察PF处理后HepG2细胞形态的变化;流式细胞仪检测细胞凋亡率。结果：PF对HepG2细胞具有显著增殖抑制作用,并呈浓度和时间依赖性;Hoechst 33258染色可见PF处理后细胞出现典型的凋亡形态学变化;流式细胞仪检测结果显示,与空白组比较,PF处理后的HepG2细胞凋亡率显著增加（P﹤0.01）。结论：PF对人肝癌细胞系HepG2细胞增殖具有抑制作用,且与诱导HepG2细胞凋亡有关。     

雷公藤甲素对肝癌细胞株HepG2顺铂化疗敏感性的影响

目的 观察雷公藤甲素对肝癌HepG2细胞的生长抑制作用,以及对顺铂化疗敏感性的影响.方法 应用不同浓度雷公藤甲素和顺铂单独或联合作用于体外培养的肝癌HepG2细胞不同时间,MTT方法 观察药物对细胞生长抑制作用;Western blot方法 分析细胞胞核中核转录因子κB(NF-κB)p65蛋白表达变化;流式细胞术检测细胞凋亡;Caspase-3活性检测试剂盒分析其凋亡机制.结果 雷公藤甲素对HepG2细胞的增值有明显生长抑制作用,呈剂量和时间依赖性;联用顺铂与单用药比较,明显提高细胞生长抑制率和凋亡率(P<0.05);HepG2细胞胞核可见NF-κB p65蛋白少量表达,单用顺铂后能使其表达增强,联用雷公藤甲素后可逆转此现象;各药物组细胞中Caspase-3活性显著增高,且联合用药组高于单用药组(P<0.05).结论 雷公藤甲素对肝癌HepG2细胞有明显的生长抑制作用,呈剂量和时间依赖性,并能增强顺铂化疗敏感性.其机制可能与抑制胞核内NF-κB p65蛋白表达和增加Caspase-3活性有关.     

Studies on uptake, sub-cellular trafficking and efflux of antisense oligodeoxynucleotides in glioma cells using self-assembling cationic lipoplexes as delivery systems

The cellular uptake of antisense oligodeoxynucleotides (ODNs) may be enhanced by the use of carriers such as cationic liposomes or lipoplexes, but little is known about the intracellular fate and subcellular trafficking of these systems in target cells. In this study, we report on the cellular uptake and biodistribution of ODNs in the presence and absence of optimised self-assembled cationic lipoplexes using the C6 glioma cell line as an in vitro model. Biotin or radiolabelled 15-mer phosphorothioate (PS) ODNs were synthesised and their cellular uptake and subcellular biodistribution characterised in the presence and absence of an optimised cationic lipoplex delivery system using studies ranging from cellular association, cellular efflux and transmission electron microscopy (TEM). Ultrastructural studies clearly showed PS ODNs in the absence of liposomal delivery to be sequestered within endosomal and lysosomal vesicular bodies indicative of endocytic uptake. ODNs were also visible, to a lesser extent, in the nucleus and cytoplasm. By employing DOSPA (2'-(1",2"-dioleoyloxypropyldimethyl-ammonium bromide)-N-ethyl-6-amidospermine tetra trifluoroacetic acid) and DOPE (dioleoylphosphatidylethanolamine) complex in a 3 : 1 ratio, as a delivery system for ODNs at a optimal lipid/DNA charge ratio of 1 : 1, the level of ODN cellular association was significantly increased by approximately 10-12 fold with a concomitant change in subcellular distribution of PS ODN. TEM studies indicated enhanced penetration of ODN within the cytosol and the cell nucleus with reduced presence in vesicular compartments. Efflux studies confirmed that cationic lipoplexes promoted entry of ODNs into 'deeper' cellular compartments, consistent with endosomal release. Optimised cationic lipoplexes improved cellular delivery of ODNs by enhancing cell association, uptake and by favourably modulating the intracellular trafficking and distribution of ODNs into non-vesicular compartments including the cytosol and nucleus.     

Hepatocellular carcinoma targeting effect of PEGylated liposomes modified with lactoferrin

A hepatocellular carcinoma targeting lactoferrin (Lf) modified PEGylated liposome system was developed for improving drug efficacies to hepatic cancer cells. In this present work, PEGylated liposomes (PLS) were successfully prepared by the thin film hydration method combined with peglipid post insertion. Lf was covalently conjugated to the distal end of DSPE-PEG2000-COOH lipid by amide bound and loaded onto PEGylated liposomes surface as the targeting ligand. To confirm the targeting efficacies to hepatic cancer, coumarin-6 and DiR were encapsulated as fluorescent probes. The confocal microscopy and flow cytometry demonstrated that Lf conjugated PEGylated liposomes (Lf-PLS) were efficiently associated by HepG2 cells, while limited interaction was found for liposomes modified with a negative control protein. A similar pharmacokinetic behavior was observed in pharmacokinetics study of the liposomal formulations. Meanwhile, the in vivo imaging of liposomes in HepG2 tumor bearing mice indicated that Lf-PLS achieved more accumulation in tumor compared with PLS without Lf conjugated. The significant in vitro and in vivo results suggested that Lf-PLS might be a promising drug delivery system for hepatocellular carcinoma therapy with low toxicity.     

Gel and solid matrix systems for the controlled delivery of drug carrier-associated nucleic acids

In order to achieve a sustained pharmacological activity of oligonucleotides (ODNs) and avoid repeated administrations, we have developed a new concept of delivery system that combine sustained release and improved intracellular penetration. These systems are designed for the intravitreal delivery of antisense ODNs. The first concept consisted in using liposomes dispersed in a thermosensitive gel (poloxamer 407). After intravitreal administration in a rabbit model, liposomes and liposomes-gel formulations provided, 1-day postinjection, significantly higher drug levels than the control solution of the oligothymidilate pdT16. In addition, there was no significant difference in the amounts of pdT16 found in the vitreous humor between the liposomes and liposomes-gel. Nevertheless, because of their better stability in the absence of poloxamer, liposomes alone allowed to a larger extent to control the delivery of ODNs as compared to liposome-gel formulations since 37% of the ODNs were still found in the vitreous 15 days after administration. In addition, the ODNs found in the vitreous humor were protected against degradation by their encapsulation within liposomes. The second approach consisted in designing microspheres allowing to release in a controlled fashion pdT16. The ODN was encapsulated within poly(lactide-co-glycolide) microspheres alone or associated with polyethylenimine (PEI) at different nitrogen/phosphate (N/P) ratios. The introduction of PEI in the internal aqueous phase resulted in a strong increase of the ODN encapsulation efficiency. PEI affected microsphere morphology inducing the formation of very porous particles yielding to an accelerated release of pdT16. Porosity and controlled delivery was prevented by introducing sodium chloride in the external preparation medium. When incubated with HeLa cells, microspheres encapsulating pdT16/PEI complexes allowed an improvement of the intracellular penetration of the released ODN. Both liposomes and microspheres are suitable for local delivery of ODNs.     

油茶皂苷通过内质网应激途径诱导人肝癌细胞HepG2凋亡的研究

目的探讨油茶皂苷诱导人肝癌细胞HepG2凋亡的作用及其作用机制。方法采用MTT法考察了油茶皂苷对肿瘤细胞增殖的影响;用Western blot方法分析油茶皂苷对caspase-3、PARP、Bip、ATF6、IRE1、Perk、eIF2a和eEF2蛋白质表达的影响。结果油茶皂苷(10～30 mg.L-1)明显抑制HepG2细胞的增殖。同时可激活Caspase信号通路,诱发PARP底物的断裂;进一步上调IRE1表达量及活化Perk、eIF2a和eEF2蛋白。结论油茶皂苷通过内质网应激途径诱导人肝癌细胞HepG2发生凋亡。     

四嗪二甲酰胺抑制肝癌细胞株HepG2增殖并诱导细胞凋亡的研究

目的观察四嗪二甲酰胺(ZGDHu-1)体外抑制肝癌细胞株HepG2增殖并诱导细胞凋亡作用。方法将不同浓度的ZGDHu-1与HepG2细胞在体外培养,用台盼蓝染色、MTT法、5′-溴-2′脱氧尿苷(B rdu)-ELISA法观察ZGDHu-1对HepG2细胞增殖的抑制作用;用细胞形态学、DNA凝胶电泳、DNA含量及细胞周期分析、Annexin-V/PI双标记、Ho-echst33258荧光染色和ELISA法测定DNA片段等技术检测细胞凋亡。结果ZGDHu-1能抑制HepG2细胞增殖和活力,呈现作用时间和剂量的量效关系。HepG2细胞与ZG-DHu-1作用后,大部分细胞阻滞于G2-M期;出现典型的细胞形态改变,DNA片断化,亚G1峰检出并增加,Annexin V+/PI-表达升高,细胞内DNA片段含量增加,Hoechst33258荧光染色后出现凋亡细胞的特征性改变等均证实ZGDHu-1能诱导HepG2细胞凋亡。结论ZGDHu-1能抑制HepG2细胞增殖,并可诱导其细胞凋亡。     

败酱草单萜环烯醚酯类对HepG2、MCF7细胞增殖及凋亡的影响

目的 探讨败酱草单萜环烯醚酯类（patrinia monoterpene iridoid ether esters,PMIEE）对HepG2和MCF7细胞增殖抑制和凋亡的影响。方法 HepG2和MCF7细胞经PMIEE作用后,采用CCK8法检测细胞增殖抑制情况;Annexin V-FITC/PI双标记流式细胞术检测细胞凋亡及周期情况;细胞划痕实验检测细胞迁移状况;Western blot法检测Bcl-2、Bax、caspase3、cdc2和CyclinB1的表达情况。结果 CCK8、划痕实验和Annexin V-FITC/PI流式细胞术检测显示,PMIEE对HepG2和MCF7细胞均有显著的增殖抑制、促凋亡率和降低迁移率作用（P<0.05）,呈一定量效关系,且PMIEE对HepG2细胞的周期阻滞以G2/M期为主,MCF7细胞以G0/G1期为主;Western blot结果显示,PMIEE可显著下调2种细胞Bcl-2、cdc2、CyclinB1的表达,上调Bax和caspase3的表达水平。结论 PMIEE可诱导HepG2和MCF7细胞增殖抑制和凋亡,下调Bcl-2、cdc2和CyclinB1表达及上调Bax和caspase3表达,其抗癌的潜在机制可能与此有关。