EGF及其受体flt对荷瘤鼠喉癌细胞增殖及凋亡的影响

目的　探讨VEGF及其受体flt对荷瘤鼠喉癌细胞增殖及凋亡的影响。方法　首先建立荷瘤 (HEP 2喉癌细胞 )裸鼠动物模型 ,将抗VEGF抗体及抗VEGF受体flt抗体分别注射荷瘤鼠体内 ,通过活体标记BrdU的方法 ,观察抗VEGF抗体及抗flt抗体对喉癌HEP 2细胞增殖的影响 ;用Tunel法观察对喉癌细胞凋亡的影响。结果　抗VEGF及抗flt抗体组HEP 2细胞BrdU标记阳性率显著小于对照组 (P <0 .0 5 ) ,细胞凋亡指数则显著高于对照组 (P <0 .0 5 )。结论　以抗体中和VEGF及封闭VEGF的结合位点均可抑制喉癌细胞的增殖 ,促进喉癌细胞凋亡。VEGF与喉癌细胞的生长密切相关。     

VEGF及其受体flt对荷瘤鼠喉癌细胞增殖的影响

目的;观察血管内皮生长因子（ＶＥＧＦ）对喉癌细胞增殖动力学的影响。方法：首先建立荷瘤（ＨＥＰ－２喉癌细胞）裸鼠动物模型,将抗ＶＥＧＦ抗体及抗ＶＥＧＦ受体ｆｌｔ抗体分别注射荷瘤鼠体内,通过活栓标记ＢｒｄＵ的方法,观察抗ＶＥＧＦ抗体及ｆｌｔ抗体对喉癌ＨＥＰ－２细胞增殖的影响。结果：与对照组相比较,抗ＶＥＧＦ抗体治疗组ＨＥＰ－２细胞ｂｒｄＵ标记阳性率显著降低;     

抗VEGF抗体及抗flt抗体对人喉癌HEP-2细胞系的生长抑制作用

目的：观察抗ＶＥＧＦ抗体及抗ｆｌｔ抗体对ＨＥＰ－２人喉癌细胞生长的影响,探讨ＶＥＧＦ对ＨＥＰ－２细胞的作用及作用方式。方法：将不同浓度的抗ＶＥＧＦ及抗ｆｌｔ抗体分别加入ＨＥＰ－２细胞的培养液中,培养５天后测ＨＥＰ－２细胞的活性（ＭＴＴ法）。结果：不同浓度的ＶＥＧＦ抗体作用于ＨＥＰ－２细胞时,浓度从１０μｇ／ｍｌ至１μｇ／ｍｌ时,对ＨＥＰ－２细胞的生长具有显著的抑制作用（Ｐ〈０．０１）,当ＶＥＧＦ浓     

膜受体介导的细胞凋亡与细胞增殖

目的:观察膜受体介导的人类细胞凋亡与细胞增殖,阐明膜受体介导的细胞凋亡与细胞增殖的关系。方法:用肿瘤坏死因子-α、抗Fas抗体分别处理指数生长期的Molt-4、Jurkat以及健康人外周血淋巴细胞;应用SubG1法和Annexin V/PI等方法通过流式细胞仪检测细胞增殖和细胞凋亡。结果:处于静止期的外周血淋巴细胞对凋亡诱导剂不敏感,凋亡率是6%~8%。Molt-4、Jurkat细胞以及加入PHA刺激的外周血淋巴细胞经肿瘤坏死因子-α、抗Fas抗体诱导后出现了明显的细胞凋亡,凋亡率是15%~28%。经肿瘤坏死因子-α、抗Fas抗体诱导后,进入细胞周期增殖的细胞较静止期的细胞凋亡率有明显增加。结论:膜受体介导的细胞凋亡与细胞增殖具有协同作用。     

奈特液对荷瘤鼠寿命影响

奈特液对荷瘤鼠寿命影响贾光，白新生，孙克任（北京医科大学公共卫生学院劳动卫生教研室北京１０００８３山东新华制药厂淄博２５５００５山东医科大学卫生系毒理教研室济南２５００１２）本研究选择体重１８─２２ｇ健康昆明种小鼠６０只，随机分成六组，每组１０只。一...     

喉癌组织中c-jun蛋白的表达及其对细胞增殖和凋亡作用的研究

背景与目的:c-jun是早期原癌基因,其蛋白产物为AP-1(activatingprotein-1)转录因子的成分之一,参与许多生长因子和细胞因子基因的转录。最近人们发现肿瘤细胞内c-jun蛋白的表达水平和活性异常升高,其起着调控细胞增生、生存和凋亡的作用。本研究通过检测c-jun蛋白在喉癌组织中的表达并计算表达指数,分析其与临床因素的关系,探讨c-jun蛋白对喉癌细胞的调控作用。方法:收集我院耳鼻喉科手术切除的52例喉鳞状细胞癌组织、15例声带息肉组织和10例喉正常组织标本进行免疫组化研究,观察c-jun蛋白在这3种标本中的定位与表达。结果:喉癌组织中c-jun蛋白的表达指数犤(56.41±24.8)%犦不但明显高于声带息肉犤(32.48±17.8)%犦和喉正常组织(无表达),而且与喉癌组织分化程度和颈淋巴结转移密切相关(P均<0.01),但是与临床分期无相关性(P>0.05)。结论:c-jun蛋白在喉癌细胞中表达水平明显增高,其表达指数与喉癌组织分化程度、颈淋巴结有无转移有关。     

BTG1对喉癌细胞增殖和凋亡的影响及其机制研究

背景与目的：在多种细胞中B细胞易位基因1(B-cell translocation gene 1,BTG1)能够抑制细胞增殖,促进细胞凋亡,调节细胞周期进程及分化。该研究通过体外实验探讨BTG1高表达对喉癌Hep-2细胞增殖、凋亡及细胞周期的影响及其相关作用机制。方法：构建pEGFP-N1-BTG1,培养并转染喉癌Hep-2细胞,分为实验组(转染pEGFP-N1-BTG1的Hep-2细胞)和对照组(转染pEGFP-N1空质粒的Hep-2细胞)。采用蛋白［质］印迹法(Western blot)检测两组细胞中BTG1蛋白的表达水平;应用MTT法检测细胞的增值活性;使用流式细胞术检测细胞周期分布和磷脂酰丝氨酸外翻分析(Annexin Ⅴ-FITC/PI)检测细胞凋亡;采用Western blot法检测细胞周期调控蛋白Cyclin D1、凋亡相关蛋白Bcl-2表达情况。结果：成功构建pEGFP-N1-BTG1,Western blot检测结果显示,实验组细胞中BTG1蛋白表达水平明显高于对照组细胞(0.921±0.091 vs 0.308±0.047,P<0.05)。实验组与对照组细胞相比,从第24 h实验组细胞生长速度减慢,细胞增值能力降低,两组比较差异有统计学意义(P<0.05);实验组细胞中Cyclin D1蛋白表达水平下降(0.436±0.023 vs 0.916±0.092,P<0.05),细胞周期的G₀/G₁期细胞比例升高［(85.1±5.2)% vs (63.8±3.1)%,P<0.05)］;S期细胞比例降低［(8.3±1.1)% vs(23.1±1.5)%,P<0.05］;实验组细胞Annexin Ⅴ增多,细胞早期凋亡率升高［(10.3±1.1)% vs (2.8±0.3)%,P<0.05］,抗凋亡蛋白Bcl-2表达水平降低(0.167±0.009 vs 0.834±0.084,P<0.05)。结论：BTG1高表达能明显抑制喉癌Hep-2细胞的生长增殖、诱导凋亡,其可能的机制与BTG1参与细胞周期调控、诱导细胞凋亡相关。     

Toll样受体8在人宫颈癌细胞株HeLa中的表达及其激动剂对HeLa细胞增殖和凋亡的影响

目的 观察Toll样受体8(TLR8)在人官颈癌细胞株HeLa中的表达,探讨TLR8激动剂CL075对HeLa细胞增殖和凋亡的影响。方法 采用实时荧光定量聚合酶链反应(PCR)法,检测13种肿瘤细胞系中TLR8 mRNA和经CL075作用后HeLa细胞中环氧化酶2(COX-2)、Bcl-2和血管内皮生长因子(VEGF) mRNA的表达水平;采用免疫荧光技术对HeLa细胞中TLR8蛋白的表达进行定位观察;采用流式细胞术检测不同浓度CL075作用下HeLa细胞的细胞周期和凋亡的变化;采用四甲基偶氮唑蓝(MTT)法检测HeLa细胞的增殖状态。结果 与其他肿瘤细胞株相比,HeLa细胞中TLR8mRNA的表达水平最高,达703.7±20.6。TLR8蛋白主要定位于HeLa细胞的胞浆中。0.1、0.5和1.0 μg/ml的CL075分别作用于HeLa细胞48 h后,G2/M+S期细胞所占的比例逐渐增高,其中1.0μg/ml CL075作用组G2/M+S期细胞所占的比例最高,达(57.67±1.73)％,明显高于空白对照组[(39.02±2.33)％,P＜0.01]。经不同浓度的CL075处理后,HeLa细胞的凋亡水平与空白对照组相比,无明显改变(P＞0.05);但经顺铂处理后,凋亡细胞明显增多(P＜0.01)。MTT法检测结果显示,与空白对照组比较,CL075作用于HeLa细胞48和72 h后,HeLa细胞的增殖能力明显增强(P＜0.01)。CI075作用于官颈癌HeLa细胞24和48 h后,COX-2、Bcl-2和VEGF mRNA的表达水平明显升高(P＜0.05)。结论宫颈癌HeLa细胞中TLR8的表达水平及其与配体相互作用的信号,可能是肿瘤发生和发展的重要因素之一。TLR8可能是宫颈癌治疗的一个潜在靶点。     

分割放疗对鼻咽癌组织细胞增殖凋亡的影响

目的：探讨常规分割放疗和超分割放疗对鼻咽癌组织细胞增殖凋亡的影响。方法：采用ＴｄＴ酶介导的生物素化ｄｕｔｐ缺口末端标记技术（ＴＵＮＥＬ）和免疫组织化学Ｓ－Ｐ法,分别检测６０例鼻咽癌患者在放疗第２周和第４周时细胞凋亡率（ａｐｏｐｔｏｓｉｓ ｒａｔｅ,ＡＲ）和增殖细胞核抗原（ｐｒｏｌｉｆｅｒａｔｉｏｎ ｃｅｌｌ ｎｈｕｃｌｅａｒ ａｎｔｉｇｅｎ,ＰＣＮＡ）的表达。结果：常规分割放疗组放疗第４周后的ＡＲ     

缺氧对乳腺癌和宫颈癌裸鼠移植瘤EGFR表达及细胞凋亡的影响

目的 在乳腺癌、宫颈癌裸鼠成瘤模型中观察缺氧对表皮生长因子受体(EGFR)表达和细胞凋亡的影响。方法 以人乳腺癌MCF-7和宫颈癌HeLa移植裸鼠模型为研究对象,利用激光共聚焦显微镜观察缺氧区程度和EGFR表达情况;利用TUNEL染色观察EGFR表达对缺氧肿瘤细胞凋亡的影响。结果 在乳腺癌MCF-7和宫颈癌HeLa细胞缺氧程度高的区域,EGFR高表达和低表达均存在。此外,与EGFR的低表达肿瘤组织相比,凋亡程度在EGFR高表达肿瘤组织中降低。结论 乳腺癌、宫颈癌细胞缺氧对EGFR表达呈非均一性作用,缺氧诱导EGFR表达与细胞凋亡呈负相关。     

生酮饮食对人结肠癌裸鼠皮下移植瘤生长的影响

  目的  观察生酮饮食对人结肠癌裸鼠皮下移植瘤生长的影响; 探讨生酮饮食可能的作用机制。   方法  24只雄性BALB/C裸鼠皮下注射人结肠癌HCT116细胞系后随机分成2组, 分别给予正常饮食(standard diet, SD)及生酮饮食(ketogenic diet, KD), 两组饮食均不限制总量。当肿瘤体积达到600~700 mm3时实验终止, 并将接种当日至肿瘤达到目标体积的时间定义为肿瘤生长期。比较不同饮食对裸鼠重量、血糖、血酮体、血胰岛素水平以及肿瘤生长等的影响。   结果  与SD组相比, KD组裸鼠肿瘤生长明显延缓, KD组及SD组皮下移植瘤达到目标体积的时间分别为(33.8±6.7)天和(24.8±3.1)天。KD组裸鼠血酮体水平显著升高, 血胰岛素水平轻度下降, 而血糖水平无明显变化, KD组肿瘤组织坏死面积明显增大, 而血管密度则显著降低。   结论  不限制总量的生酮饮食可明显延缓裸鼠人结肠癌细胞皮下移植瘤模型的生长。生酮饮食抗肿瘤治疗的作用机制及其对肿瘤其他特性如侵袭性和转移性等的影响有待进一步研究证实。      

Effect of estrogens and antiestrogens on growth of human breast cancer cells in athymic nude mice

Endocrine therapy with estrogen deprivation or with antiestrogens results in tumor regression in a subset of patients with advanced breast cancer. To better understand the mechanisms by which estrogens and antiestrogens modulate breast cancer growth in vivo, we have studied the effects of endocrine manipulation on the development and growth of tumors derived from cultured human breast cancer cells in the athymic nude mouse. MCF-7 breast cancer cells were inoculated into 6-week-old female BALB/c athymic nude mice. Tumor growth did not occur in ovariectomized mice. Cells remained viable, however, since estrogen supplementation more than 30 days later resulted in tumor formation. Minimal tumor growth was observed in intact female nude mice which have low circulating estrogen levels. Tumor development and growth in ovariectomized or intact mice supplemented with 17 beta-estradiol in the form of a s.c. pellet were dose dependent; growth rates increased with estrogen doses ranging from 0.01 to 0.5 mg. Antiestrogen treatment with either tamoxifen or LY156758 caused transient stimulation of tumor growth, followed by a prolonged stationary phase. Growth resumed with estrogen supplementation. Treatment of mice bearing established MCF-7 tumors with estrogen withdrawal (removal of estrogen pellet) resulted in cessation of tumor growth, but not in tumor regression. Growth inhibition was also observed with antiestrogens and was dose dependent. However, tumor regression did not occur, even in mice treated with high doses of tamoxifen (serum concentration of 1.0 microM) for as long as 60 days. Tumor growth was restored in these mice with estrogen replenishment. Tumor cells also remained viable histologically despite prolonged (1 month) estrogen deprivation or antiestrogen therapy, although the mitotic index was markedly reduced. Similar observations were made with mice inoculated with the hormone-responsive ZR75-1 human breast cancer cells, but not with hormone-independent MDA-231 cells which were not influenced by estrogen or antiestrogen treatment. In summary, development and growth of MCF-7 and ZR75-1 tumors in nude mice are estrogen dependent. Endocrine therapy by estrogen deprivation or antiestrogen treatment inhibits tumor cell proliferation in nude mice, but does not cause tumor regression or loss of cell viability.     

白桦脂酸对人宫颈癌裸鼠移植瘤的抑制作用及机制

目的 探讨白桦脂酸（BA）对人宫颈癌HeLa细胞株裸鼠皮下移植瘤生长的抑制作用及其机制。方法 将4×10⁶个HeLa细胞接种于裸鼠右肩背部皮下,建立人宫颈癌裸鼠皮下移植瘤模型,24只荷瘤裸鼠随机分为BA高剂量（80mg／kg）、中剂量（40mg／kg）、低剂量（20mg／kg）组及空白对照组（含溶剂）,每组6只,隔日腹腔注射连续给药21d,停药24 h后处死裸鼠,测量裸鼠体重、移植瘤体积、瘤重,计算抑瘤率;光镜下观察移植瘤组织形态学变化;TUNEL法检测肿瘤细胞凋亡指数;免疫组化法检测瘤组织内caspase-3、CytoC蛋白表达。结果 裸鼠处死后,BA处理组的移植瘤体积均明显小于空白对照组（P＜0.01）,其中高、中、低剂量组抑瘤率分别为56.2％、40.4％和24.6％ （P＜0.01）;HE染色法显示BA处理组小鼠的移植瘤组织出现明显凋亡及坏死改变;TUNEL检测对照组和BA低、中、高剂量组的凋亡指数分别为（8.36±2.78）％、（20.98±3.01）％、（28.74±4.77）％和（39.34±6.15）％（P＜0.05）;免疫组化法示BA处理组的肿瘤组织caspase-3、CytoC蛋白表达水平较空白对照组明显升高（P＜0.01）。结论 BA对人宫颈癌HeLa细胞移植瘤的生长具有明显的抑制作用,其机制可能与上调caspase-3、CytoC蛋白的表达及诱导细胞凋亡有关。     

Effect of antisense VEGF cDNA transfection on the growth of chronic myeloid leukemia K562 cells in vitro and in nude mice

To further elucidate the role of vascular endothelial growth factor (VEGF) in the pathogenesis of chronic myeloid leukemia (CML), we transfected K562 cells with a VEGF(121)cDNA sense vector (S), an antisense (AS) vector or vector (V) alone. The growth of transfected cells was investigated by MTT and colony-formation assays, and apoptosis was measured by flow cytometry (FCM) of Annexin-V-FITC/PI dual labeled cells. Transfected cells were subcutaneously transplanted into nude mice and the microvessel density (MVD) of tumor masses was determined by vWF immunohistochemistry staining. We tested the supernatant of different transfected K562 cells against human bone marrow endothelial cells (BMECs), and examined the synergic effects of antisense VEGF(121)cDNA and IFNalpha or STI571 on the proliferation and apoptosis of K562 cells. We found that K562/AS transfectants exhibited a 49% reduction in VEGF secretion, whereas K562/S transfectants exhibited a 3-fold increase in VEGF secretion, all in comparison to the vector controls. K562 cells transfected with antisense VEGF(121)cDNA showed growth retardation in vitro. In transplanted nude mice in vivo, transfection of implanted cells with antisense VEGF(121)cDNA resulted in decreased tumor MVD, and increased apoptosis in the presence of IFNalpha. Taken together, these results suggest that VEGF may be involved in the pathogenesis of CML through autocrine and paracrine mechanisms, and that anti-VEGF therapy alone or in combination with conventional treatment may be beneficial for CML patients.     

VEGF receptor antisense therapy inhibits angiogenesis and peritoneal dissemination of human gastric cancer in nude mice

The efficacy of a phosphorothioate antisense oligonucleotide (ASO) for KDR/Flk-1 (KDR/Flk-1-ASO), an endothelial cell-specific vascular endothelial growth factor (VEGF) receptor, was investigated on the peritoneal dissemination and angiogenesis of a human gastric cancer cell line in nude mice. Green fluorescent protein (GFP)-transduced NUGC-4 (NUGC-4-GFP) human gastric cancer cells were implanted into the peritoneal cavity of nude mice. KDR/Flk-1-ASO, -SO, or phosphate-buffered saline was administrated from days 7 to 14, 200 microg/mouse, once a day. The mice were sacrificed on day 28. Disseminated peritoneal tumor nodules expressing GFP were visualized by fluorescence microscopy. KDR/Flk-1-ASO significantly decreased the extent of peritoneal dissemination of the tumors. The number of cells undergoing apoptosis was significantly increased in the KDR/Flk-1-ASO-treated tumors. Microvessel density was significantly reduced in the KDR/Flk-1-ASO-treated tumor nodules. The KDR/Flk-1 antisense strategy, therefore, decreases tumor dissemination apparently by inhibiting angiogenesis.     

Lewis y高表达对人卵巢癌RMG-I细胞裸鼠体内致瘤性及移植瘤VEGF和VEGFR表达的影响

目的：比较α1,2岩藻糖转移酶基因转染前后卵巢癌细胞株RMG-I的裸鼠体内致瘤性及移植瘤组织血管内皮生长因子及其受体VEGFR1和VEGFR2表达的变化。方法：利用已建立的Lewisy抗原稳定高表达卵巢癌细胞株RMG-I-H及转染前细胞株RMG-I为细胞模型,将转染前后细胞株种植裸鼠皮下建立人卵巢癌裸鼠移植瘤模型,观察两组肿瘤生长情况。第5周处死动物,测量移植瘤重量,免疫组织化学方法检测肿瘤组织VEGF及VEGFR1、VEGFR2的表达。结果：转染组及未转染组裸鼠均有荷瘤形成,但转染组的成瘤时间（5.2±0.8d）早于未转染组（8.8±1.3d）,且转染组的瘤重和体积与未转染组相比均明显增加（P〈0.05）。转染组裸鼠移植瘤组织VEGF及VEGFR2表达量明显高于未转染组（P均〈0.05）。结论：Lewisy抗原高表达能增加卵巢癌RMG-I细胞的体内致瘤性,上调裸鼠移植瘤VEGF及VEGFR2的表达。提示Lewisy抗原能提高癌细胞的恶性程度,且该作用与VEGF及VEGFR2表达增高关系密切。     

Lidamycin induces apoptosis of B-cell lymphoma cells and inhibits xenograft growth in nude mice

OBJECTIVE To study the cytotoxicity of Lidamycin （LDM） and its induction of apoptosis in Raji and Daudi cells of B-cell lymphoma, and the inhibition of growth of the lymphoma Raji xenograft in nude mice. METHODS MTT assay was used to observe the inhibition by LDM on the proliferation of the Raji and Daudi ceils. Annexin V-FITC/PI double-stain, in combination with flow cytometry （FCM）, was used to determine the induction of apoptosis by LDM in Raji cells. The B-cell lymphoma Raji xenograft model in nude mice was set up to detect the in vivo antitumor activity of LDM. RESULTS LDM markedly inhibited the proliferation of the Raji and Daudi cells in vitro, with IC50 values of 7.13×10^-11 mol/L and 2.91×10^-10 mol/L, respectively. The apoptotic rates of Raji cells were respectively 77.98% and 67.63% at 0.5 nmol/L and 0.25 nmol/L of LDM, indicating an obvious induction of apoptosis in Raji cells. LDM inhibited the formation and growth of human B-cell lymphoma Raji xenograft in nude mice. The inhibition rates of tumor growth were respectively 74.9% and 65.2% in LDM at dosage group of 0.05 mg/kg and 0.025 mg/kg, suggesting an apparent prolongation of survival time in the nude mouse bearing lymphoma. CONCLUSION LDM can effectively induce apoptosis of the B-cell lymphoma cells and inhibit the xenograft growth in nude mice.