子宫内膜癌组织中内分泌分化细胞的生物学特性及意义

观察子宫内膜癌组织中内分泌分化细胞的增殖与凋亡状况、雌激素受体(ER)与孕激素受体(PR)表达,探讨其生物学及临床意义。手术切除或活检的子宫内膜癌标本50例。采用嗜铬素A(CgA)作为内分泌分化的标记,进行CgA／PCNA及CgA/TUNEL双重染色,观察内分泌分化细胞的增殖与凋亡状况;并进行CgA与生存素(Survivin)双重免疫组化染色,探讨生存素与内分泌分化细胞凋亡的关系。通过CgA/ER及CgA/PR双重免疫组化染色观察内分泌分化细胞的ER、PR表达。结果显示CgA阳性细胞呈PCNA及TUNEL染色阴性。CgA阳性细胞多表达Survivin。CgA阳性细胞密集的区域ER或PR减少明显。大多数CgA阳性细胞不表达ER或PR。结果提示子宫内膜癌组织的内分泌分化细胞属暂不增殖细胞群,亦极少发生凋亡,是较稳定的细胞群。内分泌分化细胞高表达Survivin可能是其逃避凋亡的分子学基础。子宫内膜癌组织的内分泌分化与ER和PR的减少有关。     

Ki67反义多肽核酸抑制肾癌生长的体外及动物实验研究

为研究肿瘤增殖基因Ki67反义多肽核酸(AS-PNAs)对人肾癌细胞的体内外抑制作用.将AS-PNAs(10.0μmol/L)转染人肾癌786-0细胞,采用免疫组化、Western印迹技术检测Ki67表达;3H-TdR掺入试验检测细胞增殖;免疫组化TUNEL法检测细胞凋亡。裸鼠移植瘤内注射AS-PNAs(10.0μmol/L)连续4d后,第3、6、12天处死小鼠,取瘤组织检测肿瘤体积、Ki67表达、细胞凋亡。结果发现,AS-PNAs处理组786-0细胞Ki67表达明显降低,3H-TdR掺入率明显降低,细胞凋亡率明显增加,与随机PNAs对照组比较差异均有显著性(P<0.01)。动物实验AS-PNAs处理组小鼠肿瘤体积明显缩小,Ki-67抗原表达明显降低,细胞凋亡明显增加,与对照组比较差异均有显著性(P<0.01)。Ki67基囚AS-PNAs在体外及动物体内均有抑制增殖、促进凋亡作用,是一种有前途的反义治疗药物。     

胎儿及育龄妇女子宫内膜内分泌细胞的免疫组化研究

目的：探讨胎儿子宫内膜内分泌细胞的形态特征、意义及可能的起源。方法：12-40周胎儿子宫内膜组织20例，育龄妇女正常子宫内膜20例。采用嗜铬素A(Chromogmnin A，CgA)免疫组织化学染色标记内分泌细胞，并进行CgA与上皮性起源标记物——细胞角蛋白(Cytokeratin，CK)双重免疫组织化学染色。     

柴胡皂苷D对人肝癌HepG2细胞系增殖和裸鼠肝癌形成的抑制作用

目的：探索柴胡皂苷D(Saikosaponin-d,SSd)对人肝癌HepG2细胞系增殖和裸鼠肝癌形成的影响。方法：CCK-8检测细胞增殖;流式细胞术分析细胞凋亡;蛋白印记检测细胞Ki67和cleaved caspase-3表达;脱氧核糖核苷酸末端转移酶介导的缺口末端标记(TUNEL)染色分析肿瘤组织细胞凋亡;免疫组化检测肿瘤组织Ki67和cleaved caspase-3表达。结果：柴胡皂苷D组HepG2细胞增殖能力明显低于对照组,细胞凋亡明显高于对照组(P<0.05)。与对照组相比,柴胡皂苷D组HepG2细胞Ki67表达明显减弱,cleaved caspase-3的表达明显增强(P<0.05)。而且,柴胡皂苷D组裸鼠肿瘤体积明显小于对照组(P<0.05)。柴胡皂苷D处理可提高小鼠存活率。另外,柴胡皂苷D组裸鼠肿瘤组织细胞凋亡远远高于对照组(P<0.001)。与对照组相比,柴胡皂苷D组裸鼠肿瘤组织Ki67表达显著降低,cleaved caspase-3表达显著增强(P<0.01)。结论：柴胡皂苷D可抑制人肝癌HepG2细胞系增殖及裸鼠肝癌形成。     

良性前列腺增生组织中基质细胞的增殖和凋亡

目的：探讨良性前列腺基质增生组织中细胞增殖及凋亡状态的改变。方法：采用双抗体免疫组化方法，对6例良性前列腺增生和4例正常前列腺组织标本中平滑肌细胞的增殖状态进行研究；采用免疫组化和TUNEL相结合的方法，在连续的组织切片上观察基质细胞凋亡状态的变化。结果：在增生的前列腺组织中，处于增殖状态的平滑肌细胞比例远远高于正常前列腺组织，处于凋亡状态的基质细胞比例低于正常前列腺组织。结论：良性前列腺增生组织中，平滑肌细胞的增殖增加和基质细胞的凋亡减少同时发生。     

原代人宫颈癌细胞的培养方法

背景：在体外分离、培养宫颈癌上皮细胞, 从细胞水平及分子水平研究肿瘤病因及治疗的基础,是宫颈癌组织工程研究的最基本环节。 目的：探讨适用于组织工程宫颈癌研究的人宫颈癌原代细胞培养方法以及传代后细胞形态变化、增殖的特性。 方法：0.25%胰蛋白酶和2%Ⅰ型胶原酶联合消化法体外培养宫颈癌原代细胞,荧光倒置显微镜下观察肿瘤细胞形态、体外生长情况,免疫组化法检测体外培养宫颈癌细胞表面标记物CK17和肿瘤细胞增殖抗原Ki67表达。 结果与结论：采用胰蛋白酶与Ⅰ型胶原酶联合消化法体外分离培养人宫颈癌细胞,该法相对简单且重复性较好,所得到的原代细胞纯度较高。经体外传代培养的宫颈癌细胞仍可保持稳定的细胞表型。宫颈癌细胞表面标记物CK17阳性表达,证实细胞为上皮源性,肿瘤细胞增殖抗原Ki67表达阳性提示所培养细胞具有肿瘤细胞恶性增殖的特性。     

死亡相关蛋白激酶在卵巢上皮性癌的表达及临床意义

目的检测卵巢上皮性癌中死亡相关蛋白激酶的表达及与病理分期、细胞分化程度、淋巴转移的相关性。方法用免疫组化SP法检测正常卵巢组织、良性卵巢上皮性肿瘤及卵巢上皮性癌中DAPK的表达水平。结果卵巢上皮性癌中DAPK的阳性表达率51．61％，明显低于正常卵巢组织（100％）和良性卵巢上皮肿瘤（80．96％），P〈0．05。研究还发现DAPK低表达与卵巢上皮性癌的病理分级、细胞分化程度及淋巴转移有关。结论DAPK作为一种促细胞凋亡基因的表达产物，其表达下调与卵巢上皮性癌的发生、发展过程密切相关；DAPK可能通过促进细胞凋亡和细胞分化对卵巢上皮性癌的侵袭与转移起抑制作用。     

口腔扁平苔藓中肿瘤坏死因子α和肿瘤坏死因子受体Ⅰ表达及与细胞凋亡的关系

目的：探讨口腔扁平苔藓肿瘤坏死因子-α、肿瘤坏死因子受体Ⅰ的表达意义及其与细胞凋亡的关系。方法：采用脱氧核糖核苷酸末端转移酶介导的原位缺口末端标记法(TUNEL法)、免疫组化技术检测50例口腔扁平苔藓及10例正常口腔粘膜中肿瘤坏死因子α(TNF-α)、肿瘤坏死因子受体Ⅰ(TNFRⅠ)的表达及细胞凋亡情况。结果：口腔扁平苔藓的上皮角质细胞TNFRⅠ及固有层TNF-α表达均明显高于正常对照组，而上皮TNF-α及固有层TNFRⅠ的表达则明显少于正常对照组。口腔扁平苔藓的上皮层可见大量凋亡细胞，而固有层单个核细胞的凋亡指数则低于正常对照组。结论：口腔扁平苔藓中同时存在角质细胞的凋亡亢进及单个核细胞凋亡的减少，这种凋亡的异常可能与口腔扁平苔藓的发生发展密切相关。     

胎儿子宫内膜内分泌细胞的免疫组化观察

目的：观察胎儿子宫内膜内分泌细胞的发育规律、形态特征及含物，探讨其生物学意义。方法：12～39w胎儿子宫内膜20例，采用嗜铬素A免疫组化染色标记内分泌细胞；血清素，生长抑素及胃泌素释放肽的免疫组化染色，观察胎儿子宫内膜内分泌细胞的内含物。结果：最早在16w子宫膜内膜上皮出现CgA阳性细胞，分布在子宫上皮及腺上皮，以靠近基底处较多。各胎龄组子宫内膜均未见血清素、生长抑素及胃泌素释放肽阳性细胞。结论：胎儿子宫内膜与成人子宫内膜的内分泌细胞在分布、数量和类型上存在差异。内分泌细胞在胚胎早期出现，提示内分泌细胞对子宫内膜的发育具有特殊意义。     

Ki67在正常妊娠和胎儿生长受限患者胎盘组织中的表达和意义

目的 通过检测增殖蛋白Ki67在正常妊娠各期和FGR患者胎盘组织中的表达，探讨其在正常妊娠中的作用及与FGR的关系。方法 采取免疫组化法取正常妊娠各期和FGR患者胎盘组织共42例，检测Ki67蛋白的表达。结果 （1）Ki67主要在细胞滋养细胞中表达强，在合体滋养细胞中呈阴性。（2）Ki67在早、中、晚期胎盘组织阳性表达率无统计意义（P〉0．05），表达强度早、中期及中、晚期无显著差别（P〉0．05）；早、晚期有显著差异（P〈0．05）：FGR组与正常妊娠晚期组胎盘中Ki67表达阳性率、表达强度之间无显著差异（P〉0．05）。结论 （1）Ki67在妊娠早期、中期表达强度高，说明滋养细胞处于高增殖状态。（2）Ki67随妊娠进展表达下调，提示与胎盘老化和分娩发动有关。（3）FGR组Ki67表达阳性率上调说明细胞滋养细胞的增生。     

基因干扰CXCL4对GBC-SD细胞恶性增殖和上皮间质转化的影响

目的 探究基因干扰CXCL4对GBC-SD细胞恶性增殖和上皮间质转化的影响.方法 构建shR-NACXCL4载体转染至GBC-SD细胞,将细胞随机分为3组:Control组、shRNA-NC组、CXCL4-shRNA1组.EDU染色检测细胞增殖;流式细胞仪检测细胞凋亡;荧光显微镜观测细胞上皮间质转化形态学变化;West...     

尖锐湿疣角质形成细胞凋亡及Ki-67的表达

目的：研究尖锐湿疣（ＣＡ）角质形成细胞的凋亡及Ｋｉ－６７的表达及其相关性。方法：对４８例ＣＡ采用ＴＵＮＥＬ技术原位检测其细胞的凋亡,用免疫组化方法,观察Ｋｉ－６７阳性的表达。结果：表皮中见到典型的凋亡细胞,表皮各层中有较多Ｋｉ－６７阳性细胞。病理严重度与凋亡指数无显著差异性（Ｐ〉０．０５）;与Ｋｉ－６７阳性细胞指数比较则有显著差异性（Ｐ〈０．００５）。凋亡细胞与Ｋｉ－６７阳性细胞之间呈负相关（ｒ＝     

靶向沉默CD105和Ki67的表达对卵巢癌OVCAR3细胞生物学行为的影响

目的 探讨沉默CD105和Ki67基因表达对人卵巢上皮癌OVCAR3细胞系生物学行为的影响。 方法 CD105-siRNA、Ki67-siRNA、CD105-siRNA+Ki67-siRNA、阴性对照组分别转染卵巢癌OVCAR3细胞,用MTT法、划痕实验、Transwell小室及流式细胞术检测CD105、Ki67单基因沉默及联合基因沉默对人卵巢癌细胞增殖、迁移、侵袭及凋亡的影响。 结果 与各自的空白对照组及空脂质体对照组相比,单基因CD105-siRNA组、单基因Ki67-siRNA组和双基因联合干预组基因沉默后人卵巢癌OVCAR3细胞的增殖能力、迁移能力及侵袭能力均明显下调,其中联合干预组下降最为明显,癌细胞凋亡率明显增加,其中联合干预组增加最为明显,差异存在统计学意义（P<0.01,P<0.05）。 结论 CD105-siRNA和Ki67-siRNA表达载体均可抑制人卵巢癌OVCAR3细胞的增殖,并降低其细胞迁移和侵袭能力,诱导其肿瘤细胞凋亡。双基因联合沉默,效果更加显著。     

Apoptosis and proliferation in lungs of human fetuses exposed to chorioamnionitis

AIMS: To determine whether chorioamnionitis has an impact on the extent of apoptosis and proliferation in fetal lungs. Fetuses exposed to chorioamnionitis have an increased risk of aquiring lung tissue damage in utero. METHODS AND RESULTS: Lung tissue sections from 35 stillborn fetuses were used in this study. Chorioamnionitis-exposed fetuses were subdivided depending on whether pneumonia was diagnosed (n = 13) or not (n = 10); 12 unaffected fetuses served as controls. Apoptotic and proliferating cells were determined by in-situ terminal deoxytransferase-mediated dUTP nick end labelling (TUNEL) assay and by anti-Ki67 immunohistochemistry, and quantified. The median apoptotic index in lungs of chorioamnionitis-exposed fetuses increased 2.4-fold compared with chorioamnionitis-negative stillborn controls (P = 0.043) and rose 21.6-fold when chorioamnionitis-exposed fetuses additionally developed pneumonia (P < 0.001). Compared with the proliferation index of the control group (PI = 2.3), the median percentage of proliferating cells in the lungs of chorioamnionitis-exposed fetuses decreased (PI = 1.4) (P = 0.036), but increased 1.8-fold (P = 0.036) in fetal lungs of the chorioamnionitis/pneumonia group. By double labellings combining the TUNEL assay or the Ki67 antigen with cell marker proteins, we identified distal airway epithelial cells as the cell type undergoing apoptosis in chorioamnionitis-exposed fetal lungs, while epithelial, endothelial and smooth muscle cells proliferated. Immunolabellings of cleaved caspases -8 and -9 revealed that apoptosis is mediated via initiator caspase-8. CONCLUSION: Chorioamnionitis induces apoptosis of distal airway epithelial cells via the caspase-8 pathway and interferes with the normal proliferative activity of epithelial, endothelial, and smooth muscle cells in fetal lungs. Thus, apoptosis and proliferation are an important feature of chorioamnionitis-associated lung injury in utero.     

Homeostatic mass control in gastric non-neoplastic epithelia under infection of Helicobacter pylori: an immunohistochemical analysis of cell growth, stem cells and programmed cell death

We evaluated homeostatic mass control in non-neoplastic gastric epithelia under Helicobacter pylori (HP) infection in the macroscopically normal-appearing mucosa resected from the stomach with gastric cancer, immunohistochemically analyzing the proliferation, kinetics of stem cells and programmed cell death occurring in them. Ki67 antigen-positive proliferating cells were found dominantly in the elongated neck portion, sparsely in the fundic areas and sporadically in the stroma with chronic infiltrates. CD117 could monitor the kinetics of gastric stem cells and showed its expression in two stages of gastric epithelial differentiation, namely, in transient cells from the gastric epithelial stem cells to the foveolar and glandular cells in the neck portion and in what are apparently progenitor cells from the gastric stem cells in the stroma among the infiltrates. Most of the nuclei were positive for ssDNA in the almost normal mucosa, suggesting DNA damage. Cleaved caspase-3-positive foveolar cells were noted under the surface, suggesting the suppression of apoptosis in the surface foveolar cells. Besides such apoptosis of the foveolar cells, in the severely inflamed mucosa apoptotic cells were found in the neck portion where most of the cells were Ki67 antigen-positive proliferating cells. Beclin-1 was recognized in the cytoplasm and in a few nuclei of the fundic glandular cells, suggesting their autophagic cell death and mutated beclin-1 in the nuclei. Taken together, the direct and indirect effects of HP infection on the gastric epithelial proliferation, differentiation and programmed cell death suggested the in-situ occurrence of gastric cancer under HP infection.     

Absence of primary cilia in cell cycle‐arrested human breast cancer cells

Previous studies using cultured cells showed that primary cilia are present in quiescent cells, but are absent in proliferating cells. We studied here the relationship between the presence or absence of primary cilia and the cell cycle arrest of normal epithelial cells and cancer cells in the human normal breast and breast cancer tissues. In normal breast tissues, although most epithelial cells were nonproliferating as estimated by the immunofluorescence staining of the proliferation marker Ki‐67, primary cilia were present only in 20–40% of the epithelial cells. In breast cancer tissues, primary cilia were not observed in any of the breast cancer cells. Furthermore, primary cilia were hardly observed in the nonproliferating cancer cells in the orthotopic and metastatic human breast cancer xenograft tumors in mice. These results indicate that the absence of primary cilia does not necessarily represent the proliferating phases of normal epithelial cells and cancer cells.     

洛伐他汀对胶质瘤干细胞增殖和凋亡的影响

BACKGROUND:Increasing evidence has shown that lovastatin with less toxicity to normal cells has crucial effects on proliferation, apoptosis and differentiation of various cancer cells. However, its roles in glioma stem cells remain unclear. OBJECTIVE:To explore the effect of lovastatin on proliferation and apoptosis of glioma stem cells. METHODS:Flow cytometric sorting was used to separate glioma stem cells from human glioblastoma cell line U87. Effects of lovastatin on the proliferation and apoptosis of glioma stem cells were determined by MTT and flow cytometry, respectively. Furthermore, expression levels of Ki67, Bax and Bcl-2 in glioma stem cells treated with lovastatin were detected using western blot analysis. RESULTS AND CONCLUSION:The CD133-positive glioma stem cells were sorted from human glioblastoma cell line U87 with a positive percentage of 85%. MTT assay showed that lovastatin inhibited the proliferation of glioma stem cells in dose (5, 10, 20 μmol/L)- and time (24, 48, 72, 96 hours)-dependent manners. Flow cytometry analysis showed that 10 μmol/L lovastatin (48 hours) induced apoptosis in glioma stem cells. In addition, the expression level of Ki67 was decreased by lovastatin treatment in a dose-dependent manner, and the Bcl-2 and Bax expression levels were reduced and increased by 10 μmol/L lovastatin treatment, respectively. In conclusion, lovastatin can inhibit cell proliferation and induce apoptosis of glioma stem cells, and lovastatin may be a potential drug for treatment of brain tumors.