维生素E琥珀酸酯上调Fas表达诱导乳腺癌Bcap-37细胞凋亡的研究

目的检测维生素E琥珀酸酯(VES)对荷乳腺癌裸鼠的体内凋亡诱导作用。方法裸鼠皮下接种Bcap-37乳腺癌细胞建立荷瘤裸鼠模型,以150mg/kg体重剂量给予VES治疗共5周,测量肿瘤大小,以流式细胞仪检测细胞周期变化和细胞表面Fas表达,以免疫组织化学法检测肿瘤组织Fas表达并以TUNEL法检测肿瘤细胞凋亡指数的变化。结果VES对荷乳腺癌裸鼠具有显著的增殖抑制作用。细胞周期分析显示,VES治疗后肿瘤细胞表现为G_0/G_1期阻滞,凋亡率升高。乳腺癌组织Fas表达值由对照组1.40±0.55升高至治疗组3.00±0.84(P＜0.05),Fas平均荧光强度由3.28升高为5.62。肿瘤细胞的凋亡指数由5.6±1.7升高为16.6±4.6(P＜0.05)。结论VES对于荷乳腺癌裸鼠具有强效的体内生长抑制作用,VES治疗能够上调肿瘤细胞Fas表达,促进细胞凋亡。     

H2O2诱导ECV304细胞凋亡与Cyclophilin-D蛋白表达的研究

目的 探讨人脐静脉内皮细胞凋亡模型中Cyclophilin-D蛋白的表达及意义.方法 分别应用100、300、500 μmol/L H2O2按时间依从处理人脐静脉ECV3O4细胞,对照组单纯磷酸盐缓冲液(PBS)处理.检测细胞凋亡率、Cyclophilin-D蛋白及其编码基因ppif的表达量的变化.结果 500 pLmol/L H2O2处理组的ECV304细胞凋亡率、Cyclophilin-D蛋白表达量和Cyelophilin-D编码基因ppff的表达量均高于对照组(P<0.01).Cyclophilin-D表达量与H2O2作用时间和凋亡率呈显著正相关,r分别为0.967(P<0.01)和0.971(P<0.01).结论 H2O2可成功诱导血管内皮细胞氧化损伤与凋亡,Cyclophilin-D蛋白可作为细胞凋亡严重程度的预测因子.     

H2O2诱导的肝星状细胞凋亡后胶原的变化

目的 观察Ⅰ型胶原(COL Ⅰ)和Ⅲ型胶原(COLⅢ)在H2O2诱导的肝星状细胞(HSC)凋亡后的变化.方法 用流式细胞术检测100 nmol/L的H2O2诱导HSC不同程度的凋亡;应用半定量逆转录-聚合酶链反应(RT-PCR)检测细胞内COL Ⅰ、COLⅢ mRNA的表达及变化.结果 用100 nmol/L H2O2诱导HSC凋亡(凋亡率分别5.86%、58.55%、71.98%),COL Ⅰ、COL Ⅲ在活化的HSC-T6细胞内均高表达,且伴随凋亡的增加,表达急剧下降,COL Ⅰ的变化更明显.结论 诱导HSC凋亡可减少胶原基因的表达.     

H2O2诱导的肝星状细胞凋亡后胶原的变化

Objective To explore the changes of collagen (COL) Ⅰ and COLⅢ after apoptosis of HSC induced by H2O2. Methods Different apoptosis rate of HSC-T6 induced by H2O2 ( 100 nmol/L) was evaluated by flow cytometry. RT-PCR was used to detect the gene expression levels of COL Ⅰ and COL Ⅲ. Results Apoptosis of HSC was induced by 100 nmol/L H2O2 with the poptosis rate being 5.86%, 58.55% ,and 71.98% ,respectively. The COL Ⅰ and COLⅢ were highly expressed in activated HSC,and decreased sharply as apoptosis increased. The changes in gene expression of COL Ⅰ were much more obvi-ous than those of COL Ⅲ. Conclusion Inducing apoptosis of HSC may decrease the gene expression of collagens.     

H2O2诱导的肝星状细胞凋亡后胶原的变化

Objective To explore the changes of collagen (COL) Ⅰ and COLⅢ after apoptosis of HSC induced by H2O2. Methods Different apoptosis rate of HSC-T6 induced by H2O2 ( 100 nmol/L) was evaluated by flow cytometry. RT-PCR was used to detect the gene expression levels of COL Ⅰ and COL Ⅲ. Results Apoptosis of HSC was induced by 100 nmol/L H2O2 with the poptosis rate being 5.86%, 58.55% ,and 71.98% ,respectively. The COL Ⅰ and COLⅢ were highly expressed in activated HSC,and decreased sharply as apoptosis increased. The changes in gene expression of COL Ⅰ were much more obvi-ous than those of COL Ⅲ. Conclusion Inducing apoptosis of HSC may decrease the gene expression of collagens.     

H2O2诱导的肝星状细胞凋亡后胶原的变化

Objective To explore the changes of collagen (COL) Ⅰ and COLⅢ after apoptosis of HSC induced by H2O2. Methods Different apoptosis rate of HSC-T6 induced by H2O2 ( 100 nmol/L) was evaluated by flow cytometry. RT-PCR was used to detect the gene expression levels of COL Ⅰ and COL Ⅲ. Results Apoptosis of HSC was induced by 100 nmol/L H2O2 with the poptosis rate being 5.86%, 58.55% ,and 71.98% ,respectively. The COL Ⅰ and COLⅢ were highly expressed in activated HSC,and decreased sharply as apoptosis increased. The changes in gene expression of COL Ⅰ were much more obvi-ous than those of COL Ⅲ. Conclusion Inducing apoptosis of HSC may decrease the gene expression of collagens.     

H2O2诱导的肝星状细胞凋亡后胶原的变化

Objective To explore the changes of collagen (COL) Ⅰ and COLⅢ after apoptosis of HSC induced by H2O2. Methods Different apoptosis rate of HSC-T6 induced by H2O2 ( 100 nmol/L) was evaluated by flow cytometry. RT-PCR was used to detect the gene expression levels of COL Ⅰ and COL Ⅲ. Results Apoptosis of HSC was induced by 100 nmol/L H2O2 with the poptosis rate being 5.86%, 58.55% ,and 71.98% ,respectively. The COL Ⅰ and COLⅢ were highly expressed in activated HSC,and decreased sharply as apoptosis increased. The changes in gene expression of COL Ⅰ were much more obvi-ous than those of COL Ⅲ. Conclusion Inducing apoptosis of HSC may decrease the gene expression of collagens.     

H2O2诱导的肝星状细胞凋亡后胶原的变化

Objective To explore the changes of collagen (COL) Ⅰ and COLⅢ after apoptosis of HSC induced by H2O2. Methods Different apoptosis rate of HSC-T6 induced by H2O2 ( 100 nmol/L) was evaluated by flow cytometry. RT-PCR was used to detect the gene expression levels of COL Ⅰ and COL Ⅲ. Results Apoptosis of HSC was induced by 100 nmol/L H2O2 with the poptosis rate being 5.86%, 58.55% ,and 71.98% ,respectively. The COL Ⅰ and COLⅢ were highly expressed in activated HSC,and decreased sharply as apoptosis increased. The changes in gene expression of COL Ⅰ were much more obvi-ous than those of COL Ⅲ. Conclusion Inducing apoptosis of HSC may decrease the gene expression of collagens.     

H2O2诱导的肝星状细胞凋亡后胶原的变化

Objective To explore the changes of collagen (COL) Ⅰ and COLⅢ after apoptosis of HSC induced by H2O2. Methods Different apoptosis rate of HSC-T6 induced by H2O2 ( 100 nmol/L) was evaluated by flow cytometry. RT-PCR was used to detect the gene expression levels of COL Ⅰ and COL Ⅲ. Results Apoptosis of HSC was induced by 100 nmol/L H2O2 with the poptosis rate being 5.86%, 58.55% ,and 71.98% ,respectively. The COL Ⅰ and COLⅢ were highly expressed in activated HSC,and decreased sharply as apoptosis increased. The changes in gene expression of COL Ⅰ were much more obvi-ous than those of COL Ⅲ. Conclusion Inducing apoptosis of HSC may decrease the gene expression of collagens.     

E2F1蛋白在病理性瘢痕组织中的表达

目的增生性瘢痕和瘢痕疙瘩是临床上常见的病理性瘢痕,是创伤后过度愈合反应的结果,以成纤维细胞的异常增殖及合成分泌大量细胞外基质为特征,其形成机理尚不清楚,研究表明基因失调是其中的关键.E2F基因是细胞周期G1向S期过渡的重要调控因子,在调节细胞周期进程和调节细胞增殖过程中起着关键作用.本实验的目的是检测E2F1基因在病理性瘢痕组织中的表达,以正常皮肤组织做对照,初步探讨E2F1在病理性瘢痕形成中的生物学作用.方法利用免疫组化ABC法检测正常皮肤、成熟瘢痕、增生性瘢痕和瘢痕疙瘩组织中E2F1蛋白的表达,并进行统计学分析.结果增生性瘢痕和瘢痕疙瘩组织中E2F1蛋白表达两组间无明显差异,与正常皮肤、成熟瘢痕对照组比较均有显著性差异(P＜0.01).结论 E2F1蛋白表达在病理性瘢痕组织中增高,促进瘢痕组织中细胞的增生,对病理性瘢痕的形成可能起着重要作用.     

转录因子E2F1在非梗阻性无精症睾丸组织中的表达

目的旨在探讨E2F1核转录因子在人类睾丸组织中的表达与生精功能的关系。方法选取32例人类非梗阻性无精子症睾丸组织石蜡标本，采用HE染色观察曲细精管病理变化，同时用免疫组化SP法检测睾丸组织中核转录因子E2F1的表达;对照组选取13例人类正常睾丸组织。通过分析病理组HE染色，比较E2F1阳性染色范围及程度，评价E2F1核转录因子与生精功能的关系。结果非梗阻性无精子症睾丸组织HE染色显示精曲小管中无或仅少量精子，精曲小管的生精上皮脱落、排列紊乱，精曲小管壁部分有玻变、纤维变，生精细胞萎缩。32例非梗阻性无精子症睾丸组织中E2F1阳性表达8例，阴性表达24例，阳性率25%;13例正常睾丸组织中E2F1的阳性表达9例，阴性表达4例，阳性率69.23%;x2=7.694，差异有显著性（P〈0.01）。结论（1）E2F1核转录因子表达缺失与睾丸生精功能障碍密切相关;（2）非梗阻性无精子症睾丸组织精曲小管的生精上皮脱落、排列紊乱，生精阻滞。     

E2F1在肾透明细胞癌中的表达及意义

目的：探讨E2F1在肾透明细胞癌中的表达及意义。方法：通过实时定量PCR和Western blot方法检测E2F1在肾透明细胞癌及对应瘤旁组织中mRNA和蛋白的表达情况,并分析E2F1的mRNA水平与临床病理资料的关系及表达相关性。结果：与对应瘤旁组织相比,E2FJ的mRNA水平在肾透明细胞癌手术标本中表达明显升高（P=0．0002）,相应的蛋白水平对比与mRNA变化一致;E2F1的mRNA水平在不同年龄、性别组间差异无统计学意义（P〉0．05）,而在组织学分级、肿瘤直径大小、T分期、临床分期和大血管浸润与否差异有统计学意义（P〈0．05）。结论：E2F1表达上调可能在肾透明细胞癌的肿瘤形成中发挥作用,且E2F1表达升高可能促进肾透明细胞癌的恶性进展。     

E2F1蛋白在病理性瘢痕组织中的表达

目的　增生性瘢痕和瘢痕疙瘩是临床上常见的病理性瘢痕 ,是创伤后过度愈合反应的结果 ,以成纤维细胞的异常增殖及合成分泌大量细胞外基质为特征 ,其形成机理尚不清楚 ,研究表明基因失调是其中的关键。E2F基因是细胞周期G1向S期过渡的重要调控因子 ,在调节细胞周期进程和调节细胞增殖过程中起着关键作用。本实验的目的是检测E2F1基因在病理性瘢痕组织中的表达 ,以正常皮肤组织做对照 ,初步探讨E2F1在病理性瘢痕形成中的生物学作用。方法　利用免疫组化ABC法检测正常皮肤、成熟瘢痕、增生性瘢痕和瘢痕疙瘩组织中E2F1蛋白的表达 ,并进行统计学分析。结果　增生性瘢痕和瘢痕疙瘩组织中E2F1蛋白表达两组间无明显差异 ,与正常皮肤、成熟瘢痕对照组比较均有显著性差异 (P <0 .0 1)。结论　E2F1蛋白表达在病理性瘢痕组织中增高 ,促进瘢痕组织中细胞的增生 ,对病理性瘢痕的形成可能起着重要作用     

E2F-1蛋白在胃癌中的表达及意义

目的探讨E2F-1蛋白在胃癌及正常胃黏膜组织中的表达与意义。方法采用免疫组化技术对80例原发性胃癌癌组织与癌旁黏膜中E2F-1的表达进行检测，同时对40例正常胃黏膜组织进行对照性研究，并进行统计学分析。结果E2F-1在癌组织与癌旁黏膜中的阳性表达率为72．5％（58／80）和30．0％（24／80），在正常胃黏膜组织中的阳性表达率为22．5％（9／40），差异有非常显著性（P〈0．001）。癌组织、癌旁黏膜中E2F-1的表达在不同的临床病理特征之间差异无显著性（P〉0．05）。结论E2F-1蛋白在胃癌组织中超表达，E2F-1的反常表达与胃癌的发生有关。     

E2F1 expression in LNCaP prostate cancer cells deregulates androgen dependent growth, suppresses differentiation, and enhances apoptosis

INTRODUCTION AND OBJECTIVES: To investigate the role of E2F/RB in androgen independent proliferation, differentiation, and sensitivity to apoptotic stimuli of LNCaP prostate cancer cells. METHODS: The effects of E2F1 overexpression on androgen independent proliferation, differentiation, and apoptotic responses was assessed by flow cytometry, Western blot analysis and staining of nuclei. RESULTS: Overexpression of E2F1 in LNCaP cells confers resistance to an androgen withdrawal-mediated growth arrest, prevents differentiation, and modifies apoptotic responses. Androgen independent proliferation is associated with a dose dependent elevation of cyclin E. Cells expressing high levels of E2F1 continue to express androgen receptor and have a diminished expression of neuronal specific enolase when cultured in androgen-depleted media. Additionally, E2F1-expressing cells are more sensitive to etoposide-induced apoptosis. Western blot analysis revealed that LNCaP-E2F1 cells have elevated expression of p73, Apaf-1, caspase-3, caspase-7, but expression of caspase-8 and -9, p14(ARF), and Mcl-1, is unaltered. CONCLUSION: This is the first study that describes E2F1-dependent modifications of androgen dependence, differentiation, and sensitivity to apoptotic stimuli in LNCaP cells. Our analysis also identifies a subset of E2F1 targets that are instrumental in altering proliferative, differentiation, and apoptotic properties. Deregulation of the E2F/RB pathway and subsequent modification of key regulatory proteins may promote the development of hormone-refractory prostate tumors.     

E2F1在病理性瘢痕的表达与CD105标记的微血管密度的关系

目的探讨E2F1和CD105对病理性瘢痕血管生成的作用。方法采用免疫组化的方法,检测E2F1和CD105在40例病理性瘢痕,20例非病理性瘢痕以及20例正常皮肤的表达。通过计数和统计学方法探讨E2F1和CD105标记的微血管密度的关系。结果E2F1和CD105标记的微血管计数在病理性瘢痕和非病理性瘢痕、正常皮肤的表达有显著性差异。在病理性瘢痕中,E2F1的表达和CD105标记的微血管密度呈正相关。结论E2F1可能对病理性瘢痕形成过程中血管的形成起到重要的作用。     

IGF-1 protects intestinal epithelial cells from oxidative stress-induced apoptosis

BACKGROUND: Reactive oxygen species (ROS) are involved in the pathogenesis of necrotizing enterocolitis (NEC) in premature infants. We have recently found that activation of multiple cellular signaling transduction pathways occurs during ROS-induced intestinal cell apoptosis; the phosphatidylinositol 3-kinase (PI3-K) pathway plays an anti-apoptotic role during this process. Insulin-like growth factor (IGF)-1 activates PI3-K pathway to promote cell survival; however, the effects of IGF-1 treatment during gut injury are not clearly defined. The purpose of this study was to determine whether IGF-1 protects intestinal cells from ROS-induced apoptosis. MATERIALS AND METHODS: Rat intestinal epithelial (RIE)-1 cells were treated with either IGF-1 (100 nm), hydrogen peroxide (H2O2; 500 microm), or combination. Western blotting was performed to assess phosphorylation of Akt, a downstream effector of PI3-K. Cell Death Detection ELISA, DCHF, and JC-1 assays were performed to demonstrate protective effects of IGF-1. Wortmannin, an inhibitor of PI3-K, was used to show PI3-K-dependent mechanism of action for IGF-1. RESULTS: H2O2 treatment resulted in increased intestinal epithelial cell apoptosis with intracellular ROS generation and mitochondrial membrane depolarization; IGF-1 pre-treatment attenuated this response without affecting ROS production. H2O2-induced phosphorylation of Akt was further increased with IGF-1 treatment; wortmannin abolished these effects in RIE-1 cells. CONCLUSIONS: PI3-K pathway is activated during ROS-induced intestinal epithelial cell injury; IGF-1 exerted an anti-apoptotic effect during this response by PI3-K activation. A better understanding of the exact role of IGF-1-mediated activation of PI3-K may allow us to facilitate the development of novel therapy against NEC.