8-硝基白杨素抑制HL-60细胞系增殖及促凋亡

目的 观察8-硝基白杨素(NOChR)抑制人急性髓性白血病细胞株 HL-60 细胞增殖和诱导凋亡作用。方法 用MTT法检测HL-60细胞增殖活性;胎盼蓝染色细胞计数法测定细胞存活率,绘制细胞生长曲线;DNA凝胶电泳观察HL-60细胞凋亡;流式细胞术检测HL-60细胞凋亡率。结果 NOChR对HL-60细胞增殖具有抑制作用,呈浓度依赖性;NOChR显著抑制HL-60细胞生长;NOChR能有效地诱导HL-60细胞凋亡,呈浓度依赖性。结论 NOChR具有抑制HL-60细胞增殖和诱导细胞凋亡作用。     

8-溴-7-甲氧基白杨素和索拉非尼对肝癌干细胞样细胞凋亡的影响

背景：拟尝试结合8-溴-7-甲氧基白杨素和索拉非尼的优点扬长避短,弥补肝细胞癌肿瘤干细胞对索拉非尼的耐药,综合提高肝细胞癌的治疗效果。 目的：观察8-溴-7-甲氧基白杨素和索拉非尼单用及二者联用对体外培养的SMMC-7721细胞系肝癌干细胞样细胞凋亡的影响,并分析其作用机制。 方法：8-溴-7-甲氧基白杨素、索拉非尼单用作为对照组,二者合用作为实验组,分别作用于SMMC-7721细胞系球形细胞24 h,流式细胞术检测细胞凋亡情况,Western blot分析NF-κB(p65)蛋白表达水平。 结果与结论：与8-溴-7-甲氧基白杨素单用和不同浓度索拉非尼单用对照组比较,8-溴-7-甲氧基白杨素与索拉非尼(2.5,5,10,50 μmol/L)二者合用实验组明显提高了SMMC-7721细胞系球形细胞的凋亡率,且明显下调了NF-κB(p65)蛋白的表达。结果表明8-溴-7-甲氧基白杨素可增强索拉非尼对肝癌干细胞样细胞凋亡的诱导作用,作用机制与NF-κB(p65)蛋白表达下调可能相关。 中国组织工程研究杂志出版内容重点：干细胞;骨髓干细胞;造血干细胞;脂肪干细胞;肿瘤干细胞;胚胎干细胞;脐带脐血干细胞;干细胞诱导;干细胞分化;组织工程     

白杨素对人骨肉瘤细胞MG-63增殖与凋亡的影响

目的探讨白杨素对人骨肉瘤细胞MG-63增殖与凋亡的影响及可能机制。方法采用不同浓度白杨素(0,2.5,5,10,20,40 mg/L)作用于MG-63细胞后,MTT法和流式细胞仪分别检测白杨素对细胞增殖与凋亡的影响。蛋白免疫印迹检测白杨素(20 mg/L)对骨肉瘤细胞Bax、Bcl-2、Caspase-3、Survivin蛋白表达的影响。Real-time PCR方法检测白杨素(20 mg/L)对骨肉瘤细胞Bax mRNA、Bcl-2 mRNA、Caspase-3 mRNA、Survivin mRNA表达水平的影响。结果白杨素抑制MG-63细胞增殖、促进MG-63细胞凋亡,并呈浓度依赖性。经20 mg/L白杨素处理后,MG-63细胞Bax与Caspase-3蛋白与mRNA表达水平显著升高,而Bcl-2与Survivin蛋白与mRNA表达水平显著降低。结论白杨素在体外抑制MG-63细胞增殖,促进MG-63细胞凋亡,与调控凋亡相关蛋白Bax、Bcl-2、Caspase-3、Survivin的表达相关。     

ARMCX1对宫颈癌SiHa细胞增殖和凋亡的影响

目的:通过RNA干扰技术敲减含Armadillo重复序列X连锁蛋白1(Armadillo repeat-containing X-linked protein 1,ARMCX1)的表达,观察ARMCX1对人宫颈癌SiHa细胞增殖和凋亡等生物学特性的影响,为研究ARMCX1在宫颈癌发病机制中的作用提供依据。方法:敲减ARMCX1表达后,通过流式细胞术检测SiHa细胞的周期分布和凋亡的变化,利用平板技术观察敲减10 d后宫颈癌SiHa细胞的集落形成情况。利用Western blot实验检测细胞增殖和凋亡相关蛋白水平的变化。结果:敲减ARMCX1表达后SiHa细胞的集落形成能力明显受抑制(P<0.05),细胞周期阻滞在S期,细胞中细胞周期蛋白E(cyclin E)和磷酸化细胞分裂周期蛋白25A(cell division cycle 25A,Cdc25A)的蛋白水平降低。同时,敲减ARMCX1表达促进SiHa细胞的凋亡,Bcl-2蛋白表达量显著减少,Bax和active caspase-3蛋白量显著增加(P<0.05)。结论:敲减ARMCX1的表达抑制SiHa细胞增殖,诱导细胞凋...     

白杨素对离体培养致炎大鼠软骨细胞凋亡及内质网应激GRP78/PERK/CHOP通路的影响

目的：观察白杨素（CHR）对大鼠软骨细胞凋亡的影响,并探究其可能分子机制。方法：提取大鼠软骨细胞,使用脂多糖（LPS）诱导体外骨关节炎模型,将细胞随机分为对照（control）组、LPS组及CHR处理组（处理浓度分别为1和5 mg/L）;采用CCK-8法测定不同条件下软骨细胞的活力;采用流式细胞术检测细胞凋亡情况;Hoechst 33342染色法观察凋亡细胞核变化;Western blot检测各组细胞凋亡相关蛋白及内质网应激相关蛋白[葡萄糖调节蛋白78(GRP78)、转录激活因子6(ATF-6)、CCAAT/增强子结合蛋白同源蛋白（CHOP）、蛋白激酶R样内质网激酶（PERK）、p-PERK、真核起始因子2α(eIF2α)和p-eIF2α]的表达水平。结果：CHR浓度为1和5 mg/L时可显著提高LPS致炎软骨细胞的活力（P<0.05）,逆转细胞凋亡情况,上调Bcl-2的蛋白表达（P<0.05）,抑制Bax、cleaved caspase-3、caspase-9、GRP78、ATF-6和CHOP的蛋白表达（P<0.05）,降低p-PERK/PERK和p-eIF2α/e...     

3-溴丙酮酸对人卵巢癌细胞增殖和凋亡的影响

目的 观察3-溴丙酮酸对人卵巢癌SK-OV-3细胞株增殖和凋亡的影响,探讨3-溴丙酮酸对人卵巢癌细胞的体外抗肿瘤效应.方法 MTT比色法和集落形成试验检测3-溴丙酮酸对SK-OV-3细胞的增殖抑制作用;采用投射电镜观察3-溴丙酮酸作用后的SK-OV-3细胞形态学改变;流式细胞术检测3-溴丙酮酸作用后SK-OV-3细胞的凋亡,并对细胞周期的变化进行分析.结果 3-溴丙酮酸对SK-OV-3细胞的增殖有明显的抑制作用,具有时间（一段时间内）和剂量依赖性（P＜0.05）.3-溴丙酮酸主要将细胞阻滞于G0/G1,S期细胞明显减少.结论 3-溴丙酮酸对SK-OV-3细胞增殖的抑制效应具有时间依赖性（一段时间内）和剂量依赖性,并可诱导其凋亡.     

槲皮素诱导人宫颈癌Hela细胞凋亡的作用

目的:检测槲皮素体外对人宫颈癌Hela细胞的增殖抑制及凋亡诱导作用,并探讨线粒体在诱导凋亡机制中的作用.方法:采用酸性磷酸酶法检测细胞增殖抑制率;丫啶橙(AO)/溴化乙锭(EB)荧光染色观测细胞形态结构变化;流式细胞仪检测线粒体膜电位（△ψm）.结果:槲皮素对人宫颈癌Hela细胞有明显的增殖抑制作用,且呈浓度和时间依赖性.经槲皮素作用48 h后,AO/EB染色可见细胞呈凋亡形态学改变.100 μmol/L槲皮素作用Hela细胞24、48 h后,与对照组相比线粒体膜电位下降.结论:槲皮素体外能抑制人宫颈癌Hela细胞生长,引起线粒体膜电位下降,诱导细胞凋亡.     

3-甲基腺嘌呤对喜树碱诱导的宫颈癌Hela 细胞凋亡的影响

目的:本研究探讨自噬抑制剂3-甲基腺嘌呤(3-methyladenine,3-MA)对喜树碱(Camptothecin,CPT)诱导的Hela 细胞凋亡的影响。方法:用四甲基偶氮唑盐比色法(MTT 方法)检测CPT 对Hela 细胞作用的最佳药物浓度和时间,以及不同药物对Hela 细胞增殖活性的影响;用免疫印迹及免疫荧光检测不同药物作用于Hela 细胞后,Hela 细胞自噬标志蛋白微管相关蛋白1 轻链3(microtubule-associated protein 1 light chain 3,LC3)、p62 及凋亡相关蛋白的变化;DAPI 核染色观察细胞凋亡。结果:CPT 作用于Hela 细胞后,Hela 细胞增殖活性明显下降,并且可诱导自噬现象的发生。CPT 和3-MA 联合作用较单独CPT 作用Hela 细胞的增殖活性降低,细胞自噬水平下降,凋亡率明显升高。结论:CPT 在诱导Hela 细胞凋亡的同时可诱导自噬,通过3-MA 抑制自噬可增强Hela 细胞对CPT 作用的敏感性。     

人脐带间充质干细胞对宫颈癌Hela细胞增殖特性的影响

背景：间充质干细胞具有向炎症损伤部位和肿瘤组织特异性趋化的特性,深入研究间充质干细胞对肿瘤细胞生长增殖的影响成为亟需解决的问题。 目的：探讨人脐带间充质干细胞对宫颈癌Hela细胞增殖特性的影响。 方法：将不同细胞数的人脐带间充质干细胞或不同质量浓度的人脐带间充质干细胞条件培养基与宫颈癌Hela细胞共培养3 d,CCK-8检测Hela细胞的增殖抑制情况。 结果与结论：Hela细胞与人脐带间充质干细胞比例分别为1︰3、1︰2、1︰1、2︰1、4︰1、8︰1的情况下,相对应的细胞增殖抑制率分别为67.12%、47.18%、31.15%、27.61%、15.55%、15.95%;利用5,10,20,40,60,100 mg/L人脐带间充质干细胞条件培养基对Hela细胞进行培养,相对应的细胞增殖抑制率分别为0.61%、40.1%、63.47%、80.61%、93.56%、90.65%,说明人脐带间充质干细胞和条件培养基呈一定的浓度依赖性抑制Hela细胞的增殖。 中国组织工程研究杂志出版内容重点：干细胞;骨髓干细胞;造血干细胞;脂肪干细胞;肿瘤干细胞;胚胎干细胞;脐带脐血干细胞;干细胞诱导;干细胞分化;组织工程     

白杨素对人肝癌BEL-7402细胞表面超微结构和蛋白质磷酸化及相关通路的影响

背景：白杨素是保健食品和中草药中常见的具有防癌抗癌活性的食源黄酮类天然产物。白杨素是否有助于防治肝癌？其作用机制为何？是否促进肝癌干细胞的分化？对此尚缺乏系统研究。 目的：以人肝癌细胞为对象,揭示白杨素的亚细胞和分子作用机制。 方法：应用酸性磷酸酶实验、扫描电镜、免疫印迹技术探讨白杨素对BEL-7402细胞的作用及其机制。 结果与结论：白杨素显著抑制BEL-7402细胞的体外增殖,半数抑制浓度IC50为24.9 mol/L或6.3 mg/L。白杨素与PI3K-AKT途径特异性抑制剂LY294002联用具有显著的协同抗癌效应。但白杨素本身并不改变AKT、磷酸化pAKT、AKT下游分子GSK-3和磷酸化pGSK-3的表达水平,虽然高剂量下β-catenin略有下调。白杨素导致细胞表面微绒毛样突起密度明显增加并形成纳米膜粒;细胞回缩,间隙加宽;出现形态上不同于凋亡的死亡细胞。白杨素显著改变多种蛋白质的丝/苏氨酸磷酸化水平;对酪氨酸磷酸化亦有广谱下调作用,接近于表柔比星对照。白杨素对CDK1、磷酸化pCDK1和pCDK2、CDC25A、CDC25B和CD133均无明显影响,但可剂量依赖性诱导PARP-1降解。总之,白杨素对BEL-7402细胞的抑制作用并非起因于PI3K-AKT和Wnt/β-catenin通路阻断或细胞周期核心调控因子阻断,而更可能起因于生物膜功能障碍所致细胞表面超微结构和蛋白质磷酸化的大范围改变,并以非经典方式诱导肝癌细胞死亡。     

宫颈癌周边区淋巴管形态学观察及其意义

目的:观察人宫颈癌周边区淋巴管的形态、分布特点,探讨癌淋巴道转移机制。方法:将临床手术切除的宫颈癌组织,按癌组织中心区、周边区及正常区取材,采用LYVE-1免疫组化法、半薄切片光镜观察法,对宫颈癌淋巴管的形态分布和癌组织进行了形态学观察。结果:光镜下可见癌细胞已突破基底膜并向间质浸润。宫颈正常区粘膜层缺少淋巴管,癌周组织有较多的淋巴管,并可见淋巴管扩张,管壁模糊破损。统计癌周边区微淋巴管的面密度(0.25±0.06)、数密度(7.97±1.88)的结果表明均高于正常区(0.13±0.04)和(3.50±0.52()P<0.01)。结论:宫颈癌周边区淋巴管数量和形态的改变,为癌细胞进入淋巴管提供了形态学基础。     

Apoptosis in woman uterine cervix in pathologies associated with human papillomavirus

The level of apoptosis in uterine cervical tissue was evaluated in healthy women and in patients with various pathologies. No signs of apoptosis were found in unchanged stratified epithelium, condyloma latum, and condyloma acuminatum. The level of apoptosis decreased with progression of neoplastic epithelial transformations, usually no apoptosis was observed in samples of stage III cervical intraepithelial neoplasms. The development of preinvasive carcinoma was accompanied by activation of apoptotic processes most pronounced in the upper third of the epithelium. In some stage I and stage I-II cervical intraepithelial neoplasms, apoptosis and elimination of the basal layer cells caused rejection of the epithelium which can explain regression of this pathology at the initial stages. The prevalence of human papillomavirus infection directly correlated with neoplastic changes in the cervical epithelium.     

左炔诺孕酮下调survivin基因促进人子宫肌瘤细胞凋亡

目的探讨左炔诺孕酮(LNG)诱导人子宫肌瘤细胞(UtLMC)凋亡过程中survivin的表达变化。方法原代培养人子宫肌瘤细胞传代后,加入不同浓度LNG,以AO/EB双染法区分早、晚期凋亡细胞和坏死细胞;RT-PCR检测抑凋亡基因survivin mRNA的表达,Western blot测定survivin蛋白的表达。结果 10 mg/L LNG作用UtLMC后早期凋亡细胞多于阴性对照组,随着LNG剂量的增加,晚期凋亡细胞也逐渐增多,核浓聚、偏位,被染成桔红色;在10 mg/L以上LNG诱导的UtLMC凋亡中,survivin mRNA表达显著下降,蛋白表达由对照组的33.82±0.02下降至10.37±0.03(P<0.05)。结论一定浓度的左炔诺孕酮所诱导的人子宫肌瘤细胞凋亡,可能与抑制survivin抗凋亡活性相关。     

罗格列酮对人子宫肌瘤细胞凋亡和雌激素受体表达的影响

目的:探讨罗格列酮对人子宫肌瘤细胞增殖和凋亡的影响及可能机制.方法:不同浓度的罗格列酮(10-8,10-7和10-6 mol/L)处理原代培养的人子宫肌瘤细胞24,48和72 h.用四甲基偶氮唑盐(methyl thiazolyl tetrazolium,MTT)法测定细胞生长曲线,用流式细胞术测定细胞凋亡率,采用放射...     

Bak基因对Hela细胞的细胞周期和细胞凋亡的调控作用

目的 研究Ｂｃ１－２基因家族促凋亡蛋白Ｂａｋ在细胞凋亡细胞周期调控中的作用,评价它们作为肿瘤治疗的潜在靶基因的可能性。方法 采用一种可诱导性表达系统,（ＭＴ－Ⅱ调节系统）,在外加锌离子（ＺｎＳＯ４,１００μｍｏｌ／Ｌ）的条件下诱导Ｂａｋ基因表达。以Ｈｅｌａ细胞系作为靶细胞,获得表达Ｂａｋ基因的稳定转染子。结果 可诱导性Ｂａｋ基因过表达的Ｈｅｌａ细胞出现广泛死亡。ＴＵＮＥＬ染色证实细胞核碎片化,提示     

Smear cytology findings of large cell neuroendocrine carcinoma of the uterine cervix

Large cell neuroendocrine carcinoma (LCNEC) of the uterine cervix is a rare tumor. Moreover, there are only three reports to date that have focused on the cytologic findings of cervical LCNEC. We report the case of a 59‐year‐old Japanese woman with cervical LCNEC combined with small cell carcinoma (SmCC). Cytologic specimens from the uterine cervix demonstrated large cells with coarse chromatin and prominent nucleoli. Frequent mitotic figures were also observed. Curettage of the uterine endometrium revealed an endometrioid adenocarcinoma with squamous differentiation; i.e., an adenoacanthoma. Histologic examination of surgically resected uterine cervical tissue revealed LCNEC with minor foci of SmCC. Neuroendocrine differentiation in LCNEC was confirmed by immunohistochemistry for synaptophysin and CD56. Cytotechnologists or pathologists need to consider a differential diagnosis of LCNEC while examining cervical cytologic specimens; therefore, it is important to correctly identify the cytologic characteristics of this tumor. Diagn. Cytopathol. 2013. © 2011 Wiley Periodicals, Inc.     

Perivascular epithelioid cell tumor of the uterine cervix identified on a conventional cervical smear

Perivascular epithelioid cell tumors (PEComas) most frequently involve the uterus, particularly the uterine corpus and very occasionally the cervix. One case of PEComa identified using a conventional cervical smear has previously been documented. Herein, we present the second such case. The patient was a 51‐year‐old woman with abnormal genital tract bleeding. Samples collected for conventional cervical smears were submitted for cytopathological examination, which revealed discohesive monotonous tumor cells showing epithelioid morphology, ample cytoplasm that was pale to weakly eosinophilic, and mildly enlarged nuclei. The cytopathological features were well correlated with histopathological findings. Upon immunohistochemistry, the tumor cells were positive for both melanocytic and smooth muscle markers. Based on these findings, PEComa was diagnosed. Subsequently, a total hysterectomy with bilateral salpingo‐oophorectomy was performed, revealing that the tumor (28 × 22 × 12 mm) was located at the superficial part of the endocervix. We propose that the cytopathological findings described herein can guide the diagnosis of PEComa, even though this tumor is rare. Diagn. Cytopathol. 2015;43:1011–1016. © 2015 Wiley Periodicals, Inc.     

Cyclic mechanical stretch augments both interleukin-8 and monocyte chemotactic protein-3 production in the cultured human uterine cervical fibroblast cells

Intensive local leukocyte infiltration in the uterine cervix is a characteristic feature in the process of cervical ripening. The infiltrated leukocytes include neutrophils, macrophages and monocytes, which are believed to play important roles in cervical ripening by secreting elastase, matrix metalloproteinase and interleukin-1 (IL-1). Interleukin-8 (IL-8) and monocyte chemotactic protein-3 (MCP-3) belong to the CXC and CC chemokine families, and mediate the chemotaxis of neutrophils and monocytes/macrophages respectively. The aim of the present study was to investigate the possible involvement of IL-8 and MCP-3 in leukocyte chemotaxis in cervical ripening. Immunohistochemistry and RT-PCR detected both IL-8 and MCP-3 expression in human pregnant uterine cervices. Labour-like cyclic mechanical stretch for 48 h significantly elevated both IL-8 (555%) and MCP-3 (360%) secretion from cultured human uterine cervical fibroblast (CxF) cells (P<0.05 for both). Cyclic mechanical stretch for 24, 36 and 48 h significantly increased both IL-8 and MCP-3 mRNA expression in CxF cells (P<0.05 for all). The stretch-induced augmentation of both IL-8 and MCP-3 expression was significantly suppressed by an activator protein-1 (AP-1) inhibitor, curcumin. These data suggest that cyclic mechanical stretch of the uterine cervix by the presenting part of the fetus during labour may augment both IL-8 and MCP-3 production in the uterine cervix via AP-1 activation.     

Mixed papillary adenocarcinoma and transitional cell carcinoma of the uterine cervix

A mixed papillary adenocarcinoma and transitional cell carcinoma (MAcTcc) was discovered in the uterine cervix of a 38-year-old woman. A condylomatous papillary lesion was found in the uterine cervix during a colposcopic study and histopathological examination showed that the tumor was composed of two different neoplastic subtypes. One was an adenocarcinoma (AC) component showing papillary and tubular structure with endocervical and intestinal differentiation; the other was a transitional cell carcinoma (TCC) component showing papillary excrescence mimicking papillary TCC of urothelial origin. To characterize the tumor, an immunohistochemical study of cytokeratins (CK) was performed. The AC component showed immunoreactivities similar to conventional adenocarcinomas: positive immunoreactivity of low-molecular-weight cytokeratins 7, 8 and 19, and negative immunoreactivity of CK20 and high-molecular-weight cytokeratin (34βE12). The lower epithelial layer of the TCC component showed different immunoreactivity, but the superficial epithelial layer had similar immunohistochemical findings to the AC component. These findings indicate that the TCC component had the cellular character of AC rather than that of TCC or squamous cell carcinomas. This is thought to be the first report of a MAcTcc of the uterine cervix.