凋亡素体外诱导表达Survivin膀胱癌细胞凋亡效果观察

目的:观察凋亡素对表达Survivin膀胱癌细胞株的作用效果,并结合相应机制探讨其临床意义.方法:利用免疫组织化学法检测膀胱癌细胞株BIU-87中Survivin的表达水平,并计算平均每高倍镜视野中阳性细胞比率.将肿瘤细胞分为单纯细胞组、转染空质粒pCDNA3组及转染pCDNA3/Apoptin组,利用脂质体转染法将真核表达栽体pCDNA3/Apoptin及pCDNA3空质粒转染入膀胱癌细胞中,并于转染后24h通过RT-PCR法检测细胞中Apoptin的表达情况.分别于转染后24、48、72h利用MTT法检测各组细胞存活率,并利用流式细胞仪TUNEL法检测各组细胞的凋亡率.结果:利用免疫组织化学法检测发现膀胱癌细胞株BIU-87中Survivin呈高表达状态,平均每个高倍镜视野中阳性细胞比例约70%,并有较多细胞呈现高表达状态.转染后24h通过RT-PCR法可见Apoptin在膀胱癌细胞中表达.分别于转染后24、48、72h利用MTT法检测各组细胞存活率,发现转染pCDNA3/Apoptin组肿瘤细胞存活率均明显低于对照组,差别有统计学意义(P<0.05);利用流式细胞仪TUNEL法检测各组细胞的凋亡率,发现转染pCDNA3/Apoptin组肿瘤细胞凋亡率均明显高于对照组,差别有统计学意义(P<0.05).结论:凋亡素可在体外诱导高表达Survivin的膀胱癌细胞株BIU-87高效凋亡,结合相关机制表明凋亡素可在一定程度上克服Survivin的抗凋亡作用,因此有可能成为Survivin拮抗的抗肿瘤药物的协同用药选择.     

凋亡抑制基因存活素在膀胱癌组织中的表达及意义

目的 探讨凋亡抑制基因存活素(Survivin)在膀胱癌组织中的表达及其意义.方法 应用免疫组化(SP)法,对60例膀胱移行细胞癌组织标本中存活素的表达进行检测,并分析其表达与膀胱移行细胞癌临床病理特征的关系.结果 60例膀胱癌组织中存活素阳性表达率为65.00%,10例正常膀胱组织存活素表达均为阴性.Ⅰ、Ⅱ、Ⅲ级膀胱癌组织中存活素阳性表达率分别为37.50%、68.96%、86.66%,Ⅰ级与Ⅱ、Ⅲ级比较均有显著性差异(P＜0.05),T1、T2、T3期膀胱癌组织中存活素阳性率分别为54.54%、70.00%、85.71%,呈逐渐升高趋势,但组间比较均无显著性差异(P＞0.05).复发性膀胱癌存活素阳性率为75.00%.结论 存活素与膀胱癌发生发展有关,检测膀胱癌组织中存活素表达,对判断膀胱癌预后有一定意义,可以作为判断膀胱癌发生发展、有无复发的指标之一.     

三氧化二砷对膀胱癌BIU-87细胞增殖及生存素表达的影响

目的：探讨三氧化二砷（arsenic trioxide,As2O3）对膀胱癌BIU-87细胞增殖及生存素表达的影响。方法：用As2O3处理膀胱移行细胞癌株BIU-87后,用四甲基偶氮唑蓝法（MTT）检测细胞增殖;用细胞流式仪检测细胞周期及凋亡,生存素（Survivin）表达的变化。结果：三氧化二砷作用BIU-87细胞后,细胞生长受到明显的抑制,Survivin的表达明显减少,凋亡细胞百分比较对照组明显增多,且均具有浓度依赖性。结论：As2O3对人膀胱癌细胞株BIU-87有抑制其生长、增殖和诱导凋亡的作用,且呈现浓度依赖性。     

三氧化二砷对膀胱癌BIU-87细胞增殖及生存素表达的影响

目的:探讨三氧化二砷(arsenic trioxide,As2O3)对膀胱癌BIU-87细胞增殖及生存素表达的影响。方法:用As2O3处理膀胱移行细胞癌株BIU-87后,用四甲基偶氮唑蓝法(MTT)检测细胞增殖;用细胞流式仪检测细胞周期及凋亡,生存素(Survivin)表达的变化。结果:三氧化二砷作用BIU-87细胞后,细胞生长受到明显的抑制,Survivin的表达明显减少,凋亡细胞百分比较对照组明显增多,且均具有浓度依赖性。结论:As2O3对人膀胱癌细胞株BIU-87有抑制其生长、增殖和诱导凋亡的作用,且呈现浓度依赖性。     

改良型TAT-凋亡素体外抗人膀胱肿瘤活性的研究

目的：探讨改良型TAT-凋亡素 （TAT-VP3） 融合基因在不同人膀胱癌细胞中的表达及诱导凋亡的效应。方法： 利用改良型TAT-VP3原核表达载体pGEX-6p-1-TAT-VP3构建真核表达载体PcDNA3-TAT-VP3, 经BamHⅠ/XhoⅠ双酶切鉴定和基因测序无误后采用脂质体介导的基因转染法转染人膀胱癌BIU-87及EJ细胞, 利用RT-PCR技术检测改良型TAT-VP3基因在细胞中的表达状况。通过透射电镜扫描观察转染后不同时段肿瘤细胞的超微结构变化, MTT法检测转染重组质粒后对人膀胱癌细胞的增殖活性的影响, 流式细胞术TUNAL法检测转染重组质粒24、 48、 72h后BIU-87及EJ细胞的凋亡率, 对实验结果进行统计学分析。结果： 成功构建重组质粒PcDNA3-TAT-VP3并经测序无误, 转染人膀胱癌细胞后的RT-PCR结果证实改良型TAT-VP3基因在人膀胱癌细胞中获得表达。透射电镜观察到BIU-87及EJ细胞凋亡早期染色质聚集, 线粒体固缩, 凋亡中晚期核固缩及凋亡小体等超微形态学改变。MTT法检测结果显示转染重组质粒PcDNA3-TAT-VP3后的BIU-87及EJ细胞增殖活性明显下降 （P<0.01）。流式细胞术TUNAL法检测显示BIU-87及EJ细胞发生早期凋亡, 其凋亡率随时间的推移呈逐渐上升趋势, 转染72 h后BIU-87和EJ细胞的凋亡率分别为 （30.44±1.47） %、（42.27±2.66） %, 与对照组比较差异有统计学意义 （P<0.01）。结论： 改良型TAT-VP3基因的表达能早期、 高效地诱导不同人膀胱癌细胞发生凋亡, 为纯化改良型TAT-VP3融合蛋白以及将其应用于膀胱灌注治疗膀胱转移肿瘤的研究奠定实验基础。     

环氧化酶-2mRNA在膀胱癌中的表达及意义

目的：探讨环氧化酶-2(COX-2)mRNA在膀胱移行细胞癌(TCCB)组织中的表达及其意义。方法：采用RT-PCR方法检测78例TCCB组织中COX-2的表达，16例正常黏膜作为对照。结果：16例正常组织无COX-2表达。COX-2表达阳性率为35．9％(28／78)，COX-2表达与TCCB分级、分期之间呈正相关，P<0．05。COX-2的表达在复发者中的表达率高于未复发者，P<0．05。COX-2的表达在转移者中的表达率高于未转移者．P<0．05。结论：COX-2在TCCB的发生发展中起重要的正性调节因子的作用。COX-2与TCCB的恶性程度和进展恶化有关。COX-2可作为判断TCCB预后的指标。     

凋亡素及其诱导肿瘤细胞凋亡的作用

凋亡素是一种来源于鸡贫血病毒的小分子蛋白,能够选择性地诱导肿瘤细胞和转化细胞的凋亡,而对正常细胞无作用.凋亡素的肿瘤细胞特异性与其在细胞中的核定位密切相关.它诱导的细胞凋亡既不依赖于p53,也不会被Bcl-2的过表达所抑制,但涉及Caspase-3的激活.因此,凋亡素可作为一种新型的肿瘤诊断工具和候选抗肿瘤治疗制剂.     

细胞增殖凋亡及血管生成在肾盂输尿管移行细胞癌中的作用

我们通过检测肾盂输尿管移行细胞癌中Ｋｉ 6 7、细胞凋亡、ＣＤ31的表达 ,探讨了细胞增殖、凋亡及血管生成在肾盂输尿管移行细胞癌中的作用。一、资料与方法1 研究对象 :肾盂输尿管移行细胞癌 4 2例中 ,Ｔ1期 14例 ,Ｔ2期 17例 ,Ｔ3～ 4期 11例。细胞分级为 :1级 8例 ,2级2 2例 ,3级 12例。2 细胞增殖、微血管密度和原位凋亡细胞的检测 :用免疫组化ＳＰ法检测Ｋｉ 6 7与ＣＤ31的表达 ,用ＴＵＮＥＬ法检测凋亡细胞。在每张切片阳性细胞最密集的区域中 ,计数 10 0 0个肿瘤细胞 ,并按Ｋｉ 6 7阳性细胞所占的比率计算增殖指数(ＰＩ) ;按ＴＵＮ…     

72例膀胱移行细胞癌中Fas和FaslL的表达

目的：探讨细胞凋亡基因Ｆａｓ和ＦａｓＬ在膀胱癌保作用及与肿瘤生物学行为和预后之间关系。方法：应用免疫组化方法检测１６例正常膀胱粘膜和７２例膀胱移行细胞癌标本中Ｆａｓ和ＦａｓＬ表达。结果：膀胱癌中Ｆａｓ和ＦａｓＬ表达均显著低于正常膀胱粘 中表达；Ｆａｓ表达与肿瘤分级、分期和复发之间均密切相关，但ＦａｓＬ表达与三者之间均无相关性。结论：Ｆａｓ和ＦａｓＬ系统异常可能是膀胱癌发生发展过程中免疫逃避的重要因     

PTS(抑瘤仙)体外抗瘤作用研究

目的　研究抗瘤新药PTS(抑瘤仙 )体外对肺癌细胞、耐药肺癌细胞及对正常支气管上皮细胞的作用 ,为其临床应用提供理论依据。方法　以大细胞肺癌株NCI H 460、耐药大细胞肺癌株H 460 /cDDP及正常支气管上皮细胞株 16HBE为实验对象 ,采用 2 4h活细胞跟踪法、MTT法及台盼蓝拒染记录细胞生长曲线法 ,进行PTS的体外抗瘤作用研究 ,同时以DDP(顺铂 )为阳性对照 ,PBS(平衡盐液 )为阴性对照。结果　PTS对受到直接损伤的肺癌细胞及耐药肺癌细胞均有杀伤作用 ,具有抗瘤作用和一定的抗耐药性。与DDP比较 ,PTS有更强的抗瘤作用 ,对于H 460细胞在第 48h 5 0 0mmol的DDP杀伤率为 62 .1% ,1/2 0 0的PTS为 81.3 % ;而PTS对于正常人支气管上皮 16HBE细胞的影响较小 (DDP为 3 9.5 % ,PTS为 2 1.6% ) ,具有较高的选择性。PTS应用后立即起效并达高峰 ,作用持续时间短。结论　PTS体外抗瘤作用明显 ,并具有选择性和一定的抗耐药性     

Phytochemical Analysis and Anti-cancer Investigation of Bswellia serrata Bioactive Constituents In Vitro

Cancer is a major health obstacle around the world, with hepatocellular carcinoma (HCC) and colorectalcancer (CRC) as major causes of morbidity and mortality. Nowadays, there isgrowing interest in the therapeuticuse of natural products for HCC and CRC, owing to the anticancer activity of their bioactive constituents.Boswellia serrata oleo gum resin has long been used in Ayurvedic and traditional Chinese medicine to alleviatea variety of health problems such as inflammatory and arthritic diseases. The current study aimed to identifyand explore the in vitro anticancer effect of B. Serrata bioactive constituents on HepG2 and HCT 116 cell lines.Phytochemical analysis of volatile oils of B. Serrata oleo gum resin was carried out using gas chromatographymassspectrometry (GC/MS). Oleo-gum-resin of B. Serrata was then successively extracted with petroleumether (extract 1) and methanol (extract 2). Gas-liquid chromatography (GLC) analysis of the lipoidal matterwas also performed. In addition, a methanol extract of B. Serrata oleo gum resin was phytochemically studiedusing column chromatography (CC) and thin layer chromatography (TLC) to obtain four fractions (I, II, IIIand IV). Sephadex columns were used to isolate β-boswellic acid and identification of the pure compound wasdone using UV, mass spectra, 1H NMR and 13C NMR analysis. Total extracts, fractions and volatile oils of B.Serrata oleo-gum resin were subsequently applied to HCC cells (HepG2 cell line) and CRC cells (HCT 116 cellline) to assess their cytotoxic effects. GLC analysis of the lipoidal matter resulted in identification of tricosane(75.32%) as a major compound with the presence of cholesterol, stigmasterol and β-sitosterol. Twenty two fattyacids were identified of which saturated fatty acids represented 25.6% and unsaturated fatty acids 74.4% of thetotal saponifiable fraction. GC/MS analysis of three chromatographic fractions (I,II and III) of B. Serrata oleogum resin revealed the presence of pent-2-ene-1,4-dione, 2-methyl- levulinic acid methyl ester, 3,5- dimethyl- 1-hexane, methyl-1-methylpentadecanoate, 1,1- dimethoxy cyclohexane, 1-methoxy-4-(1-propenyl)benzene and17a-hydroxy-17a-cyano, preg-4-en-3-one. GC/MS analysis of volatile oils of B. Serrata oleo gum resin revealedthe presence of sabinene (19.11%), terpinen-4-ol (14.64%) and terpinyl acetate (13.01%) as major constituents.The anti-cancer effect of two extracts (1 and 2) and four fractions (I, II, III and IV) as well as volatile oils of B.Serrata oleo gum resin on HepG2 and HCT 116 cell lines was investigated using SRB assay. Regarding HepG2cell line, extracts 1 and 2 elicited the most pronounced cytotoxic activity with IC50 values equal 1.58 and 5.82μg/mL at 48 h, respectively which were comparable to doxorubicin with an IC50 equal 4.68 μg/mL at 48 h. Withrespect to HCT 116 cells, extracts 1 and 2 exhibited the most obvious cytotoxic effect; with IC50 values equal0.12 and 6.59 μg/mL at 48 h, respectively which were comparable to 5-fluorouracil with an IC50 equal 3.43 μg/mL at 48 h. In conclusion, total extracts, fractions and volatile oils of B. Serrata oleo gum resin proved theirusefulness as cytotoxic mediators against HepG2 and HCT 116 cell lines with different potentiality (extracts >fractions > volatile oil). In the two studied cell lines the cytotoxic acivity of each of extract 1 and 2 was comparableto doxorubicin and 5-fluorouracil, respectively. Extensive in vivo research is warranted to explore the precisemolecular mechanisms of these bioactive natural products in cytotoxicity against HCC and CRC cells.     

藤黄抗癌作用研究的回顾与展望

综述了中药藤黄 (Gamboge)及提取的有效成分在抑制肿瘤细胞、药效学和临床方面的研究资料 ,展示藤黄是一种纯天然和独创的抗癌药物 ,从而孕育着新药开发与综合使用的前景 ,值得重视     

三种药物对人乳腺癌细胞体外杀伤作用比较

目的 :比较吡柔比星(THP)和阿霉素(ADM)、表阿霉素(EPI)对人乳腺癌细胞的体外抑制及凋亡诱导作用,以及药物杀伤的时效关系。 方法 :采用ATP生物荧光肿瘤体外药敏检测技术(ATP-TCA),比较吡柔比星和阿霉素、表阿霉素对人乳腺癌细胞系MCF-7、T-47D和Bcap-37的生长抑制作用,并绘制细胞存活率-时间曲线;用Annexin V凋亡检测技术检测药物诱导细胞凋亡的情况。 结果 :3种药物对乳腺癌细胞系MCF-7、T-47D和Bcap-37的生长抑制作用呈现明显的剂量依赖性;吡柔比星对其生长抑制作用明显高于阿霉素和表阿霉素,其IC₅₀仅相当于阿霉素和表阿霉素的1/3-1/6,与后两者的差异均具有显著意义(P<0.01);阿霉素和表阿霉素的抑制作用相当,两者IC₅₀无显著性差异(P>0.05)。三种药物对乳腺癌细胞的抑制作用均呈时间依赖关系,低浓度的吡柔比星在短时间内(12h)即具有生长抑制作用,而阿霉素和表阿霉素的起效时间较为迟缓(24-36h),72h后肿瘤细胞的存活率较吡柔比星高10%-20%。3种乳腺癌细胞分别经低浓度的吡柔比星、阿霉素和表阿霉素处理后均可诱导凋亡,其中吡柔比星组的细胞凋亡比例较其它两组为高。 结论 :吡柔比星对乳腺癌细胞具有较好的生长抑制和凋亡诱导作用,其起效时间明显早于阿霉素和表阿霉素。该药良好的抑瘤效果和迅速的起效时间对于提高乳腺癌临床疗效,减轻不良反应有重要的参考和临床应用价值。     

榄香烯临床研究进展

榄香烯是温莪术抗肿瘤作用的主要活性成分,具有抗瘤谱广泛,疗效确切,毒副作用轻微,能明显提高肿瘤患者生存质量等突出优点.本文就榄香烯临床应用的现状及其亟待解决的问题作一综述.     

色素上皮衍生因子抗肿瘤治疗的研究进展

色素上皮衍生因子(pigment epithelium-derived factor,PEDF)是一种分子量大小为50kDa的分泌型糖蛋白,其具有多种生物学活性,如神经营养、神经保护、促细胞分化、抑制肿瘤生长、抑制新生血管形成等.PEDF作为一种潜在的新型抗肿瘤药物,已在不同类型肿瘤细胞的体内外实验得到证实.本文主要就PEDF的结构和功能、抗肿瘤治疗的机制及其相关的实验研究做一综述.     

全反式维甲酸抗肿瘤作用的研究进展

 维甲酸类药物包括维生素A的天然及人工合成的衍生物。维生素A（视黄醇）进入人体后转变成视黄醛，再经氧化变成维甲酸。维甲酸是维持生长发育不可缺少的物质，尤其在促进上皮组织分化生长及维持其正常功能方面起重要作用。     

In Vitro Lethal Effect of Photodynamic Therapy on Human Pancreatic Cancer Cells and Its Major Influencing Factors

OBJECTIVE To investigate the in vitro lethal effect of photodynamic therapy (PDT) using the photosensitizer hematoporphyrin on the human pancreatic cancer cell line Panc-1, the major influencing factors and the mechanisms of treatment. METHODS Three factors-the time needed for photosensitizer and cell incubation, the photosensitizer concentration (PhoC) and the exposure dose (ExpD)-were examined with different levels of these factors. Optical density (OD) was used as a measure of CCK-8 in the experiment, and was converted to the rate of cell survival. The separate effect of each factor on the photodynamic action was studied, and the interactions were investigated. The effects of different incubation times and PhoC levels on the fluorescence intensity (FI) of the intracellular photosensitizer were determined,and the mechanisms of these factors leading to the therapeutic effects of PDT discussed.RESULTS An increase in the photosensitizer and cell incubation time, an increase of PhoC, and enhancement of the ExpD, produced a corresponding decrease in the rate of Panc-1 cell survival after PDT (P , 0.05). PDT achieved its maximum lethal effects 16 h after starting the incubation, with a PhoC of 10 mg/L and an ExpD of 20 J/cm2; at these levels a synergistic interaction between PhoC and the ExpD occurred, decreasing the cell survival rate (P , 0.05). Neither simple administration of photosensitizer without ExpD (0 J/cm2) or illumination in the absence of PhoC (0 mg/L) affected the rate of cell survival (P.0.05). With an increase of PhoC and lengthening of the incubation time, the FI of the intracellular photosensitizer accordingly increased (P.0.05), and attained its maximum value at a PhoC of 10 mg/L and 36 h after the incubation. With an increase of PhoC,the FI of the photosensitizer, hematoporphyrin, in the solution increased progressively at first and then decreased (fluorescence quenching).CONCLUSION PDT with the photosensitizer hematoporphyrin has clear lethal effects on the human pancreatic cancer cell line Panc-1,but the presence of a photosensitizer and laser irradiation by themselves do not have independent lethal effects. The three influencing factors-the time for photosensitizer and cell incubation,PhoC and ExpD-correlate positively with the PDT response,within certain limits. Beyond these limits, the PDT response does not significantly increase. The main mechanism of the PDT response lies in the effect of these factors on the level of the intracellular photosensitizer and the fluorescence quenching of the photosensitizer.A synergistic effect exists between PhoC and ExpD.