赫赛汀对HER-2/neu基因高表达鼻咽癌细胞株生长的影响

目的：探讨赫赛汀(Herceptin)对HER-2／neu基因高表达鼻咽癌细胞株生长的影响及机制。方法：分别采用MTT法、^3H-TdR细胞掺入法检测Herceptin对鼻咽癌细胞株CNE-22和CNE-2体外生长的影响;用流式细胞仪检测Herceptin对CNE-2Z细胞生长、细胞周期和凋亡的影响。结果：Herceptin可抑制CNE-2Z细胞的生长,引起细胞凋亡指数的增加。结论：Herceptin可抑制高表达HER-2／neu基因鼻咽癌细胞株的生长,并诱导细胞凋亡的发生。     

赫赛汀对鼻咽癌荷瘤裸鼠肿瘤生长影响的实验研究

目的：探讨赫赛汀（Herceptin）对HER-2/neu基因蛋白高表达的鼻咽癌荷瘤裸鼠肿瘤生长的影响.方法：用鼻咽癌CNE-2Z细胞株接种于裸小鼠右腋下建立HER-2/neu基因蛋白高表达的裸鼠移植瘤模型.将已构建模型的裸鼠24只随机分为4组,每组6只,分别设阴性和阳性对照两组,Herceptin 10mg/kg和20mg/kg两实验组,以观察Herceptin对移植瘤体内生长的影响.结果：用Herceptin处理后的裸鼠移植瘤肿瘤体积均低于阴性对照组,其中10mg/kg剂量组与阴性对照组相比有统计学差异（P〈0.05）,但无量效关系;瘤重抑制率Herceptin 10mg/kg和20mg/kg分别为33.7%和23.3%,均未达到阳性标准;对裸小鼠生长无明显影响.结论：Herceptin对HER-2/neu基因过表达的鼻咽癌细胞株移植的荷瘤裸鼠肿瘤体内的生长具有一定的抑制作用,但效果并不满意.     

Herceptin联合顺铂对高表达HER-2／neu卵巢癌移植瘤鼠的治疗作用

目的：建立高表达HER-2／neu的人卵巢癌细胞株SKOV3裸鼠腹腔移植瘤模型，探讨Herceptin联合顺铂（cisplantin，cDDP）的治疗作用及其机制。方法：将人卵巢癌细胞株SKOV3接种于裸鼠腹腔，96h后随机分为4组进行腹腔内注射治疗：对照A组生理盐水，B组cDDP3mg／kg，C组Herceptin 1mg／kg，D组cDDP3mg／kg加Herceptin 1mg／kg，观察各组裸鼠成瘤率、生存期、生存延长率；采用流式细胞术检测不同治疗后肿瘤细胞中Fas、FasL的表达情况；分别取A、D组肿瘤组织予病理学检查及免疫组化染色，观察组织学变化。结果：SKOV3裸鼠腹腔移植瘤HER-2／neu免疫组化染色为＋＋，联合用药后HER-2／neu染色减弱，且凋亡现象明显。Herceptin与cDDP联合用药可明显延长荷瘤鼠的生存期，与单药相比有显著性差异（P〈0．05）。Herceptin、cDDP及联合用药后，Fas表达率均明显升高（P〈0．01），但以联合用药组表达率最高；而FasL表达率各治疗组较对照组无显著性差异。结论：Herceptin、cDDP能有效抑制高表达HER-2／neu SKOV3裸鼠腹腔移植瘤的生长，延长荷瘤裸鼠的生存期，联合用药组疗效最为显著，其机制与CD95调节有关。     

鼻咽癌患者HER-2/neu基因扩增和表达及其临床意义

目的：研究鼻咽癌HER-2／neu基因扩增、表达及其临床意义。方法：采用原位荧光杂交、逆转录多聚链式反应和免疫组化技术检测鼻咽癌组织HER-2／neu基因扩增、表达。结果：HER-2／neu基因在鼻咽癌中无扩增，但有过表达，其原因是由于mRNA高表达所致；这种过表达与鼻咽癌预后之间未显示有相关性。结论：HER-2／neu基因在鼻咽癌无扩增，有过表达，这种过表达未显示预后意义。     

Herceptin联合顺铂对高表达Her-2／neu卵巢癌体外作用的实验研究

 目的 筛选高表达Her-2／neu的卵巢癌细胞株，观察Herceptin联合顺铂（DDP）对该细胞株体外生长的抑制作用。方法 采用反转录-聚合酶链反应技术，检测人卵巢癌细胞株3AO，SKOV3，OVCAR Her-2／neu的表达情况，筛选出高表达Her-2／neu的人卵巢癌细胞株。采用四甲基偶氮唑蓝（MTT）法测定Herceptin，DDP以及联合用药对高表达Her-2／neu的人卵巢癌细胞株体外生长抑制率并进行比较。结果 人卵巢癌细胞株SKOV3 Her-2／neu呈高表达；Herceptin 可有效抑制SKOV3的生长，具有浓度依赖性（P = 0.001），联合用药组的生长抑制率为79.41 %，明显高于二者单独作用（P = 0.0001）。结论 人卵巢癌细胞株 SKOV3 Her-2／neu呈高表达。Herceptin，DDP均能有效抑制SKOV3的生长，联合用药组疗效更佳。     

亚砷酸诱导人鼻咽癌细胞中p16基因表达的研究

目的 研究亚砷酸( As2O3)诱导鼻咽癌(nasopharyngeal carcinoma,NPC)CNE- 2Z细胞株中抑癌基因p16的表达.方法 体外培养的CNE-2Z细胞分别加入不同浓度的亚砷酸,并作用于不同时间.用终浓度为2μmol/L、1 μmol/L、0.5μmol/L的As2O3加入鼻咽癌CNE-2...     

131I标记曲妥珠单抗对高表达HER-2/neu人卵巢癌的体内外实验研究

目的:观察131I 标记的曲妥珠单抗(trastuzumab,商品名为Herceptin)在体内外,对高表达人表皮生长因子受体-2(human epidermal growth factor receptor-2,HER-2)/neu的人卵巢癌细胞的靶向治疗作用.方法:采用Iodogen法用131I标记Herceptin,制备获得131I-Herceptin.实验设131I-Herceptin与顺铂(cisplatin,DDP)联用组、131I-Herceptin组、Herceptin组、DDP组及空白对照组.MTT法检测各实验组中不同药物处理对人卵巢癌SKOV3细胞的生长抑制作用.成功种植人卵巢癌SKOV3细胞移植瘤后,将荷瘤裸鼠随机分为5组,每周经尾静脉用药,连续6周,每周测量1次小鼠肿瘤的长径、短径及体质量,用药结束1周后处死小鼠,计算抑瘤率;并且采用HE染色法观察肿瘤细胞凋亡的情况,TUNEL法检测肿瘤细胞的凋亡指数;最后采用免疫组织化学法检测肿瘤组织中p53和HER-2蛋白的表达情况.结果:131I-Herceptin组的体外细胞生长抑制率及体内抑瘤率明显高于Herceptin组及DDP组;131I-Herceptin与DDP联用具有协同作用,131I-Herceptin和DDP联用组的体内肿瘤细胞凋亡指数显著高于其他各实验组,其p53和HER-2蛋白表达率则下降.结论:131I-Herceptin在体内外均能有效抑制高表达HER-2的人卵巢癌细胞生长,且与DDP联用具有协同作用.推测其可能的机制为,131I-Herceptin除影响体内HER-2蛋白表达外,还能通过内照射产生细胞毒作用,并参与p53基因调控.     

姜黄素对人鼻咽癌CNE-2Z细胞增殖及 凋亡的影响

 目的探讨姜黄素对人鼻咽癌CNE-2Z细胞增殖及凋亡的影响及其相关的分子机制。方法体外培养的 CNE-2Z细胞经不同剂量姜黄素处理后,用MTT法观察对细胞增殖的影响,流式细胞术检测细胞周期和凋亡 率的改变,Western blot检测姜黄素处理后相关蛋白Bcl-2、Bax表达水平的变化。结果姜黄素对CNE-2Z细 胞的生长和增殖都具有一定程度的抑制作用,阻断细胞于S期和G2/M期。姜黄素处理后CNE-2Z细胞凋亡抑 制基因Bcl-2蛋白表达减少,同时促凋亡基因Bax蛋白表达增加。 结论姜黄素可抑制CNE-2Z细胞增殖并促进 其凋亡。     

腺病毒E1A下调HER2/neu原癌基因表达及其抗瘤效应研究

目的　研究腺病毒E1A基因对HER2 /neu高表达肿瘤细胞生长的抑制作用及增强其化疗敏感性的作用。方法　以腺病毒Ⅴ型为载体 ,经体、内外对HER2 /neu高表达及低表达的肿瘤细胞株转染腺病毒E1A基因后 ,观察E1A基因对肿瘤细胞生长抑制作用 ;用MTT法检测E1A增强肿瘤细胞化疗敏感性作用。结果　E1A能显著抑制HER2 /neu高表达的肿瘤细胞在体内外的生长 ,延长荷瘤裸鼠的生存期。免疫印迹及免疫组织化学分析显示 ,转染E1A的HER2 /neu高表达肿瘤细胞 ,其HER2 /neu基因产物p185蛋白表达明显降低 ;经AdE1A 处理的HER2 /neu高表达的乳腺癌细胞株能明显增强对TaxotereTM的化疗敏感性。结论　E1A通过下调HER2 /neu原癌基因的表达 ,抑制HER2 /neu高表达的肿瘤细胞生长 ,增强肿瘤细胞对化疗药的敏感性。     

HER-2/neu基因在鼻咽癌中的表达及其临床意义

目的：探讨HER－2／neu基因在鼻咽癌中的表达及其临床意义。方法：采用免疫组化法测定HER－2／neu基因蛋白的表达。结果：69例鼻咽癌病人中52％（36／69）有HER－2／neu基因蛋白表达。这种过表达与肿瘤分期、局部淋巴结和远处脏器转移以及5年总生存期、3年无病生存期均无相关性。同一病例原发病灶和复发或转移灶HER－2/neu基因蛋白的表达较一致。HER－2／neu基因蛋白过表达对鼻咽癌化疗的反应性不产生影响。结论：在鼻咽癌,HER－2／neu基因蛋白过表达的临床意义不前,HER-2/neu基因蛋白能否成为治疗新的靶点还有待进一步研究。     

Anti-HER-2 anti-CD3 bi-specific antibodies inhibit growth of HCT-116 colorectal carcinoma cells in vitro and in vivo

Objective: This study is conducted to evaluate the effects of anti-HER-2×anti-CD3 bi-specific antibodies(BsAb)on HER-2/neuover-expressing human colorectal carcinoma cells. Methods: Growth was assessed by MTT assaysafter exposure of HCT-116 cells to Herceptin, anti-CD3 and BsAb antibodies. Immunocytochemistry was appliedto test the HER-2 level of HCT-116. In a nude mouse model, HER-2×CD3 BsAb was combined with effectorcells (peripheral blood lymph cells from normal human being) for observations on in Vivo growth of tumors.Results: Compared with the control group, using effector cells combined with anti-CD3 McAb, Herceptin orHER2×CD3 BsAb, tumor cell growth in vitro and in vivo was significantly inhibited (P<0.05), most remarkablyin the HER2×CD3 BsAb case. The growth of xenografts with HER2×CD3 BsAb combined with effector cells wasalso significantly inhibited when compared with the anti-CD3 McAb or Herceptin groups (P<0.05). Conclusion:HER-2/neu might be a useful target for immunotherapy in colorectal carcinoma, anti-HER2×anti-CD3 BsAbexerting clear anti-tumor effects.     

ApoG2, a novel inhibitor of antiapoptotic Bcl-2 family proteins, induces apoptosis and suppresses tumor growth in nasopharyngeal carcinoma xenografts

Nasopharyngeal carcinoma (NPC) is a common malignant tumor in South China. It has been reported that overexpression of antiapoptotic Bcl-2 family proteins in NPC has caused the lack of long-term efficacy of conventional therapies. Apogossypolone (ApoG2), a novel small-molecule inhibitor of antiapoptotic Bcl-2 family proteins, has been discovered as the optimized derivative of gossypol. In this study, we found that in NPC cells, ApoG2 totally blocked the antiapoptotic function of Bcl-2 family proteins without affecting the expression levels of these proteins. ApoG2 selectively inhibited proliferation of 3 NPC cell lines (C666-1, CNE-1 and CNE-2) that highly expressed the antiapoptotic Bcl-2 proteins. This inhibitory activity was associated with release of cytochrome c, activation of caspase-9 and caspase-3 and apoptosis of sensitive NPC cells. However, ApoG2 had no obvious inhibitory effect on NPC cell line HONE-1, which expressed antiapoptotic Bcl-2 and Bcl-xL at a low level. We further found that ApoG2 effectively suppressed tumor growth of NPC xenografts in nude mice and enhanced the antitumor effect of CDDP (cisplatin) on NPC cells in vitro and in vivo. Immunohistochemical results showed that the expression of CD31 decreased after ApoG2 treatment, which suggested inhibition of angiogenesis in NPC xenografts. Our findings strongly suggest that ApoG2 may serve as a novel inhibitor of Bcl-2 family proteins and, by targeting these proteins, may become a promising drug for the treatment of NPC.     

Induction of cell cycle arrest and apoptosis in human nasopharyngeal carcinoma cells by ZD6474, an inhibitor of VEGFR tyrosine kinase with additional activity against EGFR tyrosine kinase

ZD6474 is a vascular endothelial growth factor receptor (VEGFR) and epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor. The present study was undertaken to investigate the direct antiproliferative effect of ZD6474 on human nasopharyngeal carcinoma (NPC) in vitro and the antitumor activity on NPC xenografts in vivo. Results indicated that ZD6474 treatment inhibited EGFR phosphorylation and led to a dose- and time-dependent decrease in NPC cell (CNE-1, CNE-2 and C666-1) proliferation. Further investigation demonstrated G0/G1 cell cycle arrest in all 3 cell lines, which was associated with an upregulation of p21 and/or p27, and downregulation of CDK4, CDK6 and CDK2. ZD6474 treatment also induced apoptosis in CNE-1 and CNE-2 cells. The apoptosis mechanisms involved reduction of Bcl-2 and/or Bcl-XL, induction of Bak and/or Bax, and activation of caspases-3, -9 and/or -8. The in vivo antitumor activity was evaluated in CNE-2 and C666-1 xenografted nude mice. Administration of ZD6474 (25-100 mg/kg/day, once-daily, p.o.) produced a dose-dependent inhibition of tumor growth and prolonged survival in both models. This study suggests that ZD6474 exerts direct antiproliferative effects on NPC cell lines in vitro by inducing G0/G1 arrest and apoptosis, and potent antitumor effects on NPC xenografts in vivo. It indicates that ZD6474 may offer a new and effective treatment for human NPC.     

p53基因对鼻咽癌细胞株CNE－3裸鼠致瘤抑制作用的研究

分别将野生型和突变型ｐ５３基因导入鼻咽癌的体外培养细胞系ＣＮＥ－３．裸鼠致瘤试验表明，导入野生型ｐ５３基因的细胞系，肿瘤生长速度明显低于对照细胞系及导入突变型ｐ５３基因的细胞系；导入突变型ｐ５３基因的细胞系肿瘤生长速度最快；导入野生型ｐ５３基因的细胞系肿瘤生长速度最慢，而且肿瘤出现时间也较导入突变型ｐ５３的细胞及对照细胞晚１～２周。这说明野生型ｐ５３基因能有效地抑制肿瘤的生长，而实变型ｐ５３基因则可以促进肿瘤生长。这是首次有关ｐ５３基因对鼻咽癌细胞作用的报道。这一研究更进一步说明了ｐ５３基因的功能，对于鼻咽癌发生机理的探讨以及肿瘤的防治具有重要意义。     

转染野生型p53基因对人肺腺癌细胞作用的研究

目的研究外源性野生型p53基因对人肺腺癌细胞系GLC-82的生物学作用.方法将含有野生型p53基因的pDOR-neo逆转录病毒载体,通过Lipofectin转染GLC-82细胞,采用流式细胞分析、电镜下观察、3H-TdR掺入试验、软琼脂培养集落形成率测试及裸小鼠成瘤性实验,观察野生型p53基因对GLC-82细胞的生物学效应.结果电镜下观察可见,转染野生型p53基因可使GLC-82细胞出现一些病理现象,如细胞质中有明显的空泡及线粒体肿胀;流式细胞仪分析结果显示,野生型p53基因可阻止GLC-82细胞周期停止于G1-S区域;3H-TdR掺入试验结果提示野生型p53基因能够抑制GLC-82细胞的DNA合成,软琼脂培养集落形成率减少及裸小鼠成瘤减慢.结论转染野生型p53基因可抑制GLC-82细胞恶性增殖.     

姜黄素体内外对鼻咽癌的抗癌作用

目的研究姜黄素对鼻咽癌CNE-2Z-H5细胞株的体内外抗癌作用,探讨姜黄素在鼻咽癌治疗中的潜在应用价值。方法通过MTT法、台盼蓝拒染法绘制生长曲线观察姜黄素对CNE-2Z-H5细胞体外增殖的影响;复制CNE-2Z-H5细胞裸鼠移植瘤模型,观察姜黄素对移植瘤的体积、瘤重的影响,计算抑瘤率。结果姜黄素可以有效抑制CNE-2Z-H5细胞的增殖,并呈剂量效应关系。姜黄素可以抑制CNE-2Z-H5细胞裸鼠移植瘤的生长,低、高剂量姜黄素组的抑瘤率分别为26.19%和59.75%,且不良反应小。结论姜黄素在体内、外均可抑制CNE-2Z-H5细胞增殖,在鼻咽癌的治疗中具有一定的价值,值得进一步研究。     

Tyrosine kinase inhibitor emodin suppresses growth of HER-2/neu-overexpressing breast cancer cells in athymic mice and sensitizes these cells to the inhibitory effect of paclitaxel.

Overexpression of the HER-2/neu proto-oncogene, which encodes the tyrosine kinase receptor p185neu, has been observed in tumors from breast cancer patients. We demonstrated previously that emodin, a tyrosine kinase inhibitor, suppresses tyrosine kinase activity in HER-2/neu-overexpressing breast cancer cells and preferentially represses transformation phenotypes of these cells in vitro. In the present study, we examined whether emodin can inhibit the growth of HER-2/neu-overexpressing tumors in mice and whether emodin can sensitize these tumors to paclitaxel, a commonly used chemotherapeutic agent for breast cancer patients. We found that emodin significantly inhibited tumor growth and prolonged survival in mice bearing HER-2/neu-overexpressing human breast cancer cells. Furthermore, the combination of emodin and paclitaxel synergistically inhibited the anchorage-dependent and -independent growth of HER-2/neu-overexpressing breast cancer cells in vitro and synergistically inhibited tumor growth and prolonged survival in athymic mice bearing s.c. xenografts of human tumor cells expressing high levels of p185neu. Both immunohistochemical staining and Western blot analysis showed that emodin decreases tyrosine phosphorylation of HER-2/neu in tumor tissue. Taken together, our results suggest that the tyrosine kinase activity of HER-2/neu is required for tumor growth and chemoresistance and that tyrosine kinase inhibitors such as emodin can inhibit the growth of HER-2/neu-overexpressing tumors in mice and also sensitize these tumors to paclitaxel. The results may have important implications in chemotherapy for HER-2/neu-overexpressing breast tumors.