Aurora A激酶抑制剂VX-680对人子宫内膜癌细胞化疗敏感性的影响

目的 探讨抑制子宫内膜癌细胞中Aurora A表达与细胞对紫杉醇敏感性的相关性,明确Aurora A低表达影响紫杉醇对子宫内膜癌的治疗效果.方法 以Aurora激酶抑制剂VX-680抑制子宫内膜癌细胞株Ishikawa中的Aurora A表达,再将细胞用不同浓度紫杉醇处理,Western blot检测Aurora A蛋白受抑制情况,用流式细胞仪分析Aurora A表达下调后细胞生长抑制率和细胞凋亡率的变化.结果 Aurora A表达下调使紫杉醇时细胞的生长抑制率和细胞凋亡率均升高(P＜0.01).结论 人子宫内膜癌细胞中Aurora A表达与细胞对紫杉醇敏感性具有相关性,抑制Aurora A可望提高紫杉醇对子宫内膜癌的治疗效果.     

华蟾素对子宫内膜癌Ishikawa细胞生物学行为的影响

目的 探讨华蟾素对体外培养的子宫内膜癌Ishikawa细胞形态、增殖、凋亡、周期分布及侵袭能力的影响.方法 体外培养Ishikawa细胞,不问浓度的华蟾素干预后,光镜观察细胞形态改变,采用MTT法检测细胞的增殖,流式细胞术分析细胞凋亡及其周期,用Transwell小室检测细胞体外侵袭能力.结果 华蟾素作用后细胞由梭形、多边形变成圆形,浮起.华蟾素能有效地抑制Ishikawa细胞的增殖,降低侵袭能力,诱导细胞凋亡,并使细胞周期阻滞于G1期,减少S和G2期的细胞.华蟾素终浓度为3.0 mg/ml抑制作用最明显.结论 华蟾素抑制Ishikawa细胞增殖,降低侵袭能力,诱导细胞凋亡并阻断细胞周期.     

壳聚糖对人子宫内膜癌细胞增殖及细胞周期的影响

目的研究壳聚糖对人子宫内膜癌细胞Ishikawa的影响。方法用不同质量浓度的壳聚糖处理Ishikawa细胞48 h后,通过加入MTT和BrdU测定细胞的增殖和DNA合成情况,用流式细胞仪检测细胞周期的变化。结果壳聚糖质量浓度在不大于10μg/mL时,对Ishikawa细胞的增殖、DNA合成以及细胞周期没有明显影响。结论低质量浓度的壳聚糖（≤10μg/mL）对Ishikawa细胞的生长没有影响,可作为节育器的生物材料。     

全硫代反义hTR对人子宫内膜癌细胞生长的抑制作用

目的将互补于hTR模板区的全硫代修饰型反义寡核苷酸（PS-ASODN）作用于人子宫内膜癌HEC-1A细胞,探讨端粒酶PS-ASODN对肺癌的治疗价值。方法本实验将互补于hTR模板区的PS-ASODN转染作用于人子宫内膜癌HEC-1A细胞,然后分别采用四甲基偶氮唑蓝（MTT）比色实验、酶联免疫吸附试验（ELISA）检测人子宫内膜癌HEC-1A细胞的体外增殖、端粒酶活性、细胞凋亡的改变。结果端粒酶PS-ASODN对HEC-1A细胞生长具有明显的抑制作用。0.5～1.5μM的端粒酶PS-ASODN对HEC-1A细胞作用72h后端粒酶皆为阴性,对照组端粒酶皆为阳性。结论端粒酶PS-ASODN对体外培养的子宫内膜癌HEC-1A细胞的增殖有明显的抑制作用,其抑制作用具有浓度、时间依赖性。     

子宫内膜癌动物模型的构建

目的研究建立人子宫内膜癌的移植动物模型方法。方法将生长状态良好的Ishikawa细胞浓度调整至5×107个/0.2ml,接种于BABL/C小鼠背部皮下,观察其生长情况,留取标本行病理检查。结果成功建立了人子宫内膜癌Ishikawa细胞的BABL/C小鼠皮下移植瘤模型,形成的肿瘤具备人肿瘤生物学特点。结论建立的人子宫内膜癌Ishikawa细胞的BABL/C小鼠皮下移植瘤模型具有人子宫内膜癌特征,为研究子宫内膜癌的发病机制和药物治疗提供了实验动物模型。     

子宫内膜癌细胞中RNA干扰Bmi-1基因的实验研究及意义

目的检测子宫内膜癌细胞Ishikawa中RNA干扰Bmi-1基因的实验效果,为进一步研究奠定基础。方法培养子宫内膜癌细胞Ishikawa,采用siRNA干扰技术沉默Ishikawa细胞中Bmi-1基因,采用RT-PCR方法检测干扰后Bmi-1mRNA的表达情况,t检验用于统计学分析。结果 siRNA干扰Bmi-1后,Ishikawa细胞中Bmi-1mRNA的RT-PCR电泳条带灰度比为(0.115±0.011),阴性对照组为(0.287±0.014),两组差异具有统计学意义(P<0.05)。结论 siRNA干扰技术可以降低子宫内膜癌细胞中Bmi-1基因的表达,为进一步研究Bmi-1基因在子宫内膜癌发生发展中的作用奠定基础。     

沉默CDK7通过下调YAP表达诱导子宫内膜癌细胞凋亡

目的 研究沉默细胞周期蛋白依赖性激酶7(CDK7)诱导Yes相关蛋白（YAP）表达下调对子宫内膜癌HEC-1-A细胞凋亡的影响。方法 利用siRNA干扰技术沉默HEC-1-A细胞中CDK7的表达,采用qPCR和Western blotting检测siRNA干扰后CDK7的表达水平,CCK-8实验检测沉默CDK7对HEC-1-A细胞活力的影响,流式细胞术检测沉默CDK7对HEC-1-A细胞凋亡的影响,Western blotting检测沉默CDK7对YAP和磷酸化YAP蛋白及其下游蛋白Cyr16、CTGF表达的影响,凋亡实验检测共转染si-CDK7和pcDNA3.1-YAP对细胞凋亡的影响。结果 转染si-CDK7后,HEC-1-A细胞中CDK7 mRNA和蛋白表达水平均显著下降（P<0.01）,细胞活性显著降低（P<0.01）,细胞凋亡率显著升高（P<0.01）,YAP和磷酸化YAP蛋白及其下游蛋白Cyr16、CTGF表达水平明显下降（P<0.01）;共转染pcDNA3.1-YAP可逆转沉默CDK7导致的细胞凋亡（P<0.01）。结论 沉默CDK7可诱导Y...     

小分子干扰RNA抑制黏着斑激酶基因对子宫内膜癌细胞生物学特征的影响

目的 探讨小分子干扰RNA(small interference RNA,siRNA)抑制黏着斑激酶(focal adhesion kinase,FAK)基因对子宫内膜癌细胞生物学特征的影响。方法 培养人子宫内膜癌HEC-1A细胞,分为siRNA-FAK组、siRNA-阴性对照组和空白对照组,实时荧光定量PCR检测各组细胞中FAK基因表达,细胞增殖能力、迁移和侵袭能力检测分别采用MTT法和Transwell法,蛋白质印迹法(Western blot)检测各组细胞中FAK、磷酸肌醇3激酶(phosphoinositol 3 kinase,PI3K)、磷酸化-Akt(phosphorylate-Akt,p-Akt)、磷酸化-哺乳动物雷帕霉素(phosphorylation-mammalian rapamycin,p-mTOR)蛋白表达。结果 siRNA-FAK组细胞中FAK mRNA和蛋白相对表达量分别为(1.31±0.15)、(0.26±0.08),均低于siRNA-阴性对照组[(2.06±0.18)、(0.62±0.11)]和空白对照组[(2.11±0.20)、(0.64±0.13)],差异有统计学意义(P<0.05);siRNA-FAK组24 h、48 h、72 h、96 h时细胞吸光度A值均低于siRNA-阴性对照组和空白对照组,均差异有统计学意义(P<0.05);与siRNA-阴性对照组和空白对照组比较,siRNA-FAK组迁移细胞数和侵袭细胞数均降低,siRNA-FAK组细胞中FAK、PI3K、p-Akt、p-mTOR蛋白表达相对表达量均降低,而Akt和mTOR蛋白相对表达量均升高,均差异有统计学意义(P<0.05)。结论 沉默FAK基因表达可抑制HEC-1A细胞增殖、迁移及侵袭能力,可能与抑制PI3K/Akt/mTOR信号有关。     

钙离子调节蛋白依赖激酶在子宫内膜癌细胞中表达的研究

目的 探讨钙离子调节蛋白依赖激酶（CaMKⅣ）在子宫内膜癌发病中的作用。方法 采用免疫组化方法,检测31例子宫内膜癌患者和20例正常子宫内膜标本中CaMKⅣ的表达强度,同时分析CaMKⅣ染色细胞的百分数与患者临床特征的关系（临床分期、组织分级和临床预后）。结果 正常子宫内膜组织细胞无CaMKⅣ的表达,CaMKⅣ表达主要在子宫内膜癌细胞核内,表达的强弱与临床分期和预后有关。结论 CaMKⅣ在子宫内膜癌组织中的表达与子宫内膜癌的恶性强度有关。     

雌激素及拮抗剂三苯氧胺对子宫内膜癌细胞生长的影响

目的 研究雌激素及拮抗剂三苯氧胺对子宫内膜癌细胞HEC-1B生长的影响.方法 体外培养中分化子宫内膜癌细胞HEC-1B,按实验目的分别加入17β-雌二醇（E2,10-8M）、三苯氧胺（TAM,10-7 M）刺激24、48、72、96 h后,采用四甲基偶氮唑蓝（MTT）比色方法比较各个条件下的细胞数目.用流式细胞术方法检测加入E2、TAM后的细胞周期变化,比较各个条件下的S期细胞比率（SPF）.结果 MTT结果：E2、TAM在作用72 h和96 h后,可显著刺激HEC-1B细胞生长（P〈0.05）.HEC-1B细胞加入E2刺激后,SPF在加药72 h和96 h均增高.TAM作用细胞后,SPF值在药物作用48 h起均高于对照组.结论 雌激素、三苯氧胺均对子宫内膜癌HEC-1B细胞有促生长作用.     

Aurora激酶及其抑制剂研究状态综述

Aurora激酶家族是苏氨酸/丝氨酸激酶,在有丝分裂的染色体排列,分离和胞质分裂中起重要作用。最近的研究发现,Aurora激酶在大量人类实体瘤和血液恶性肿瘤中过表达,表明其是开发抗肿瘤药物的重要靶点。本文就近几年国内外对Aurora激酶的研究,对Aurora激酶的生物学功能、与肿瘤的关系及其抑制剂的研究进展进行综述。     

An update on the pharmacokinetics and pharmacodynamics of alisertib,a selective Aurora kinase A inhibitor

Human Aurora kinases, including Aurora kinase A (AURKA), B (AURKB), and C (AURKC), play an essential role in mitotic events such as monitoring of the mitotic checkpoint, creation of bipolar mitotic spindle and alignment of centrosomes on it, also regulating centrosome separation, bio‐orientation of chromosomes and cytokinesis. AURKA and AURKB are key regulators of mitosis and centrosome via polymerizing microfilaments and controlling chromatid segregation. In particular, AURKA plays critical roles in the regulation of mitotic entry, centrosome function, bipolar spindle assembly, and chromosome segregation. AURKA has been found to be overexpressed in various solid and haematological cancers and has been linked with poor prognosis. Its important role in cancer initiation, growth, and metastasis has brought the focus to search for potent and selective AURKA inhibitors for cancer treatment. MLN8237, also known as alisertib, is one selective AURKA inhibitor that has shown remarkable anticancer effects in preclinical studies. Alisertib exhibits favourable pharmacokinetic properties. Alisertib has generally showed good partial response rates of 4–52% and good safety profiles in Phase I and II trials when it is solely administered as well as combined with cytotoxic chemotherapeutic drugs. Recently, the multicentre, randomized Phase III study of alisertib in patients with relapsed or refractory peripheral T‐cell lymphoma has been discontinued due to unsatisfactory efficacy. The low risk of side effects, accessibility, and effectiveness of alisertib makes it a new promising anticancer therapy and further mechanistic and clinical studies are warranted.     

子宫内膜癌52例手术的远期疗效观察

目的:探讨子宫内膜癌手术切除的临床远期疗效。方法:对1998年1月—2006年12月施行手术切除52例的临床资料和随访结果进行分析。结果:1,3,5,10年总生存率分别为100%,96.2%,90.3%,79.2%。死亡共6例,死于复发转移3例,其他病死亡3例。临床分期生存率比较差异有统计学意义(P<0.01)。结论:子宫内膜癌手术治疗远期疗效满意。     

Bisphenol A affects human endometrial endothelial cell angiogenic activity in vitro

The widespread Bisphenol A (BPA) is classified as an endocrine-disrupting chemical (EDC) with estrogenic properties. Human endometrial endothelial cells (HEECs) play a key role in the endometrial angiogenesis that is under the control of estradiol. The hypothesis was that BPA may affect endometrial angiogenesis by disturbing some functional properties of the HEEC.To study this, primary HEECs were exposed to environmentally relevant doses of BPA. The HEECs were co-cultured with primary endometrial stromal cells to create conditions as similar to the in vivo situation as possible. The effects of BPA were evaluated by proliferation and viability assays, tube-formation assays, quantitative PCRs, Western blots and ELISAs.BPA slightly increased HEEC tube formation and VEGF-D protein expression compared with vehicle, without affecting HEEC viability or proliferation.Bisphenol A thus caused changes in HEEC activities in vitro, and may therefore have disturbing effects on endometrial angiogenesis.     

Aurora激酶及其抑制剂的研究进展

恶性肿瘤是威胁人类生命健康的重大难治性疾病,人类在恶性肿瘤的治疗过程中经历了漫长的历史变迁。Aurora激酶家族是苏氨酸/丝氨酸激酶,是细胞有丝分裂期重要的调节因子,可影响细胞周期进程,是抗肿瘤药物的新靶点。本文简要对Aurora激酶的生物学功能、与肿瘤的关系及其抑制剂的研究进展进行综述。     

Targeting cell cycle kinases and kinesins in anticancer drug development

The cell cycle is regulated by kinases such as the cyclin-dependent kinases (CDKs) and non-CDKs, which include Aurora and polo-like kinases, as well as checkpoint proteins. Mitotic kinesins are involved in the establishment of the mitotic spindle formation and function, and also play a role in cell cycle control. The disruption of the cell cycle is a hallmark of malignancy. Genetic or epigenetic events result in the upregulation of these kinases and mitotic kinesins in a myriad of tumour types, suggesting that their inhibition could result in preferential targeting of malignant cells. Such findings make the development of these inhibitors a rational and attractive new area for cancer therapeutics. Although challenges of potency and non-specificity have hampered their progress through the clinic, several novel compounds are presently in various phases of clinical trial evaluation.     

裂解型腺病毒Ad-E1A的构建以及体外对肝癌细胞的作用

目的 构建Ad-E1A裂解型腺病毒载体,研究其对肝癌细胞的裂解作用.方法 以QBI-293A细胞基因组DNA为模板,用特异性引物PCR法扩增E1A基因,酶切后连接到pAdTrack-CMV转移质粒上,用PmeI酶对pAdTrack-CMy-E1A线性化后,与pAdEasy-1共转化在BJ5183大肠杆菌中进行同源重组,筛选重组腺病毒质粒pAdEasy-1-pAdTrack-CMV-E1A;再用PacI酶切对重组腺病毒质粒线性化,脂质体转染QBI-293A细胞,收获Ad-E1A腺病毒子.在QBI-293A细胞中多轮感染后获得高效价的病毒.将Ad-EIA腺病毒感染SMMC-7721,并用MTT和流式细胞仪检测Ad-E1A对人肝癌细胞生长和细胞周期的影响.结果 获得高滴度的重组裂解型腺病毒Ad-E1A.Ad-E1A感染SMMC-7721后,出现明显的细胞病变.Ad-E1A对人肝癌细胞SMMC-7721的生长有明显抑制,48 h后效果明显.Ad-E1A可直接诱导SMMC-7721细胞的凋亡.结论 裂解型腺病毒Ad-E1A构建成功并可以显著抑制人肝癌细胞SMMC-7721的生长,并在肝癌细胞中复制造成细胞裂解,诱导肝癌细胞的凋亡.