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EXPERIMENTAL RESEARCH OF EFFECTIVENESS OF siRNA-her-2/neu ON DRUG SENSITIVITY OF her-2/neu-OVER-EXPRESSING LUNG ADENOCARCINOMA CELL LINE

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Chinese Journal of Cancer Research 2006年第3期18卷 173-176页

作者：任淑华王井伟曲平刘义张伟张林 Department of Thoracic Surgery The First Affiliated Hospital China Medical University Shenyang 110001 Tangshan Worker Hospital Tangshan 063000 Tumor Marker Research Center Cancer Hospital & Cancer Institute Chinese Academy of Medical Sciences Beijing 100021

Objective： her-2/neu protein overexpression has been demonstrated in lung adenocarcinoma. Its overexpression often indicates a poor prognosis and resistance to chemotherapeutic agents. The objectives of this paper is to explore the effectiveness of double-stranded short inhibitory RNAs （siRNA） targeting her-2/neu oncogene on the drug sensitivity of her-2/neu-overexpressing lung adenocarcinoma cells. Methods： Lung adenocarcinoma cell line calu-3 was transfected with siRNAs formulated LipofectAMINE 2000, and her-2/neu protein and P-gp were determined by flow cytometry （FCM）. The chemosensitivity of transfected cells to cisplatin （CDDP） was measured by MTT. Cell apoptosis detection kit （Annexin V method） was used to examine the drug induced apoptosis rate. Results： siRNA targeting her-2/neu greatly reduced the cell surface expression of her-2/neu protein and had no effect on P-gp. Consequently the inhibitory rate of CDDP in combination with siRNA targeting her-2/neu was （67.1±2.3）%, while the inhibitory rates were （48.1±3.5）%, （46.3±5.9）% and （50.2±2.9）% in untreated control, empty vector and unrelated siRNA groups, respectively. The FCM results showed that the apoptosis rate of cells treated with CDDP combined with siRNAs-her-2/neu was elevated when compared with unrelated siRNA group and empty vector group. Conclusion： Sequence specific siRNA targeting her-2/neu was capable of enhancing the chemosensitivity of calu-3 cell to cisplatin.

关键词： her-2/neu siRNA Lung adenocarcinoma cells Cisplatin

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Downregulation of wild-type p53 protein by her-2/neu mediated PI3K pathway activation in human breast cancer cells: its effect on cell proliferation and implication for therapy

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Cell Research 2004年第6期14卷 497-506页

作者： Li ZHENG, Jia Qiang REN, Hua LI, Zhao Lu KONG, Hong Guang ZHU Department of Pathology, Shanghai Medical College, Fudan University, Shanghai, 200032, China. Institute of Irradiation Medicine, Fudan University, Shanghai, 200032, China Department of Pathology Shanghai Medical College Fudan University Shanghai China Institute of Irradiation Medicine Fudan University Shanghai China

Overexpression and activation of her-2/neu (also known as c-erbB-2), a proto-oncogene, was found in about 30% of human breast cancers, promoting cancer growth and making cancer cells resistant to chemo- and radio-therapy. Wild-type p53 is crucial in regulating cell growth and apoptosis and is found to be mutated or deleted in 60-70% of human cancers. And some cancers with a wild-type p53 do not have normal p53 function, suggesting that it is implicated in a complex process regulated by many factors. In the present study, we showed that the overexpression of her-2/neu could decrease the amount of wild-type p53 protein via activating PI3K pathway, as well as inducing MDM2 nuclear translocation in MCF7 human breast cancer cells. Blockage of PI3K pathway with its specific inhibitor LY294002 caused Gl-S phase arrest, decreased cell growth rate and increased chemo- and radio-therapeutic sensitivity in MCF7 cells expressing wild-type p53. However, it did not increase the sensitivity to adriamycin in MDA-MB-453

关键词： her-2/neu PI3K breast cancer p53 MDM2

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THE EFFECT OF RNAI-MEDIATED GENE SILENCING ON her-2/neu GENE EXPRESSION IN LUNG ADENOCARCINOMA CELLS

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Chinese Journal of Cancer Research 2005年第2期17卷 84-89页

作者：任淑华王井伟曲平刘义张伟张林 Department of Thoracic Surgery The First Affiliated Hospital China Medical University Shenyang 110001 Tang Shan Worker Hospital Tangshan 063000 Tumor Marker Research Center Cancer Hospital & Cancer Institute Chinese Academy of Medical Sciences Beijing 100021

Objective: her-2/neu protein overexpression has been demonstrated in Non-small cell lung cancer, especially in lung adenocarcinoma. Its overexpression often indicates a poor prognosis. Therapeutic agents against her-2/neu have been intensively sought over the past decades. The objective of this paper is to investigate the effect of chemically synthesized siRNA targeting her-2/neu on her-2/neu dysregulated human lung adenocarcinoma cells. Methods: Calu-3 cells were transfected with siRNAs formulated LipofectAMINE 2000. The her-2/neu mRNA and protein levels were detected by RT-PCR and flow cytometry (FCM). The biological morphology and growth inhibition of calu-3 cells were observed with light microscopy and MTT assay, respectively. The cell cycle and apoptosis rate were analyzed by FCM. Results: siRNA targeting her-2/neu down-regulated the transcription of her-2/neu oncogene and protein level. Slowed cell proliferation and cell cycle arrest at G0/1 stage could be also observed, accompanied with enhanced cell apoptosis. Conclusion: Specific siRNA targeting her-2/neu can effectively inhibit her-2/neu expression in her-2/neu overexpressing calu-3 cell lines. siRNA-mediated gene silencing may be a useful therapeutic strategy for cancer.

关键词： RNAi her-2/neu Lung adenocarcinoma cells siRNA

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GENISTEIN INHIBITS EXPRESSION OF VASCULAR ENDOTHELIAL GROWTH FACTOR IN her-2/neu TRANSFECTED HUMAN BREAST CANCER MCF-7 CELLS

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Chinese Journal of Cancer Research 2006年第2期18卷 83-87页

作者：朱俊东余小平糜漫天 Department of Nutrition and Food Hygiene The Third Military Medical University Chongqing 400038

Objective： our previous studies have demonstrated that her-2/neu gene expression in human breast cancer MCF-7 cells promotes angiogenesis in MCF-7 cells xenograft tumors, and genistein inhibits angiogenesis in MCF-7 cells with her-2/neu expression xenograft tumors. here, the effects of genistein on the expression of vascular endothelial growth factor （VEGF） in MCR-7 cells with her-2/neu expression were further studied for exploring the molecular mechanism of anti-angiogenesis in her-2/neu-overexpressing breast cancer by genistein. Methods： her-2/neu-overexpressing MCF-7 cells （MCF-7/her-2） were established by transfecting HEg-2/neu gene into her-2/neu negative expression breast cancer MCF-7 cells. Immunocytochemical staining, western blot and reverse transcription-polymerase chain reaction （RT-PCR） were adopted to measure the expression of VEGF in MCF-7/her-2 cells treated by genistein for 24, 48 and 72h. Results： her-2/neu expression up-regulated VEGF mRNA and protein in MCF-7 cells, genistein decreased VEGF mRNA and protein level in MCF-7/her-2 cells in a time-dependent manner. Conclusion： These results suggest that VEGF plays an important role in her-2/neu gene expression promoted antiogenesis in breast cancer and genistein induced down-regulation of the expression of VEGF may be one of the molecular mechanisms of its anti-angiogenesis in her-2/neu-overexpressing breast cancer.

关键词： Genistein Vascular endothelial growth factor her-2/neu Breast cancer

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Correlations of β-catenin,Ki67 and her-2/neu with gastric cancer

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Asian Pacific Journal of Tropical Medicine 2014年第4期7卷 257-261页

作者： Hong-Wen Wu Cheng-Yong Qin Ji-Lai Huang Xian-Yi Kong Wen-Ji Wang Wen-Kun Bai School of Medicine Shandong University Digestive Internal Medicine The First Hospital of Zibo City Digestive Internal Medicine Shandong Provincial Hospital Digestive Internal Medicine Shandong Qianfoshan Hospital

Objective:To study the clinical pathologic characteristics ofβ-catenin,Ki67 and her-2/neu in gastric cancer and the correlation ofβ-catenin and Ki67 to the protein expression and gene conditions of her-2/***:The protein expression ofβ-catenin,Ki67 and her-2/neu was detected by immunohistochemistry in 101 cases of gastric cancer and the gene conditions of her-2/neu by fluorescence in situ hybridization(FISH).Results:The protein expression ofβ-catenin,Ki67 and her-2/neu had close relationship with the clinical pathologic characteristics of gastric ***β-catenin and Ki67 had obvious correlation to the differentiation,infiltration and lymphatic metastasis of the gastric cancer(Pher-2/neu had close relationship with the differentiation and tumor-node-metastasis staging of gastric cancer(Pher-2/neu(r=0.567,Pher-2/neu can be used as a reliable method to help the observation of biological behavior,diagnosis and prognosis of gastric cancer,and Ki67 can be used to serve the preliminary screening of her-2/neu gene state.

关键词： Gastric cancer β-catenin Ki67 her-2/neu Immunohistochemistry Fluorescence in situ hybridization Tumor marker

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her-2/neu配体结合区2蛋白的甘露糖化修饰

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细胞与分子免疫学杂志 2004年第1期20卷 95-98页

作者：徐明郭宁王红霞李爱玲胡美茹沈倍奋军事医学科学院基础医学研究所军事医学科学院仪器测试分析中心北京100850

目的 :建立一种蛋白质甘露糖化修饰的方法。方法 :用离子交换层析和钴亲和层析法 ,纯化重组her 2 /neu配体结合区2 (LBD2 )蛋白 ,并将纯化后的LBD2蛋白进行甘露糖化修饰。新合成的LBD2拟糖蛋白经质谱法检测后 ,用间苯二酚硫酸法进一步... 详细信息

目的 :建立一种蛋白质甘露糖化修饰的方法。方法 :用离子交换层析和钴亲和层析法 ,纯化重组her 2 /neu配体结合区2 (LBD2 )蛋白 ,并将纯化后的LBD2蛋白进行甘露糖化修饰。新合成的LBD2拟糖蛋白经质谱法检测后 ,用间苯二酚硫酸法进一步鉴定。结果 :纯化后LBD2蛋白的纯度可达 90 %。新合成的LBD2拟糖蛋白在质谱图上呈现相应于甘露糖修饰后蛋白质分子量的预期峰形。经间苯二酚硫酸法检测 ,结合于LBD2蛋白上的化学基团为糖分子。结论 :获得了LBD2拟糖蛋白。

关键词： her-2/neu 配体结合区甘露糖化肿瘤疫苗

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AMPLIFICATION OF THE C-erbB-2(her-2/neu) PROTO-ONCOGENE IN OVARIAN CARCINOMAS

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Chinese Medical Journal 1994年第8期 31-35页

作者：樊庆泊卞美璐刘文淑黄尚志马佳郎景和许秀英

C-erbB-2(her-2 / neu) proto-oncogene is mainly expressed in epithelial tissue and activated due to its amplification. Amplification of the C-erbB-2 proto-oncogene has been associated with poor prognosis in human ovarian cancer. Our study was to examine whether amplification is more frequently observed in ovarian cancer, or it is associated with poor prognosis of human ovarian cancer in China. The DNA of ovarian cancers was extracted and consequently digested with restriction endonuclease EcoRI, electrophoresed in 0.8% agarose gels and blotted onto nitrocellulose filter with Southern transfering method. It was then hybridized with a 32P-labelled C-erbB-2 probe and subsequently underwent autoradiography. The result has shown that the C-erbB-2(her-2 / neu) gene was amplified in 8 of 26 human ovarian cancers (30.8%). The clinical data showed that all of the 8 cases with the amplified C-erbB-2 were in their advanced stage (III-IV). Five of the patients died from 2 to 4 months after operation. These data suggest that amplification of the C-erbB-2 gene may play a role in the pathogenesis of ovarian carcinoma; it is frequently observed in advanced ovarian cancer and is associated with poor prognosis for these patients.

关键词：口口 her-2/neu CN AMPLIFICATION OF THE C-erbB-2

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her-2/neu基因在肝细胞癌中的表达及意义

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南方医科大学学报 2010年第2期30卷 326-328页

作者：张俊凯潘佩玲吴颖猛萧剑军彭杰文中山市人民医院化疗科广东中山528403

目的研究her-2/neu在肝细胞癌(HCC)患者中的表达及其临床意义。方法应用免疫组织化学SP法检测30例HCC、10例肝硬化、10例正常肝组织中her-2蛋白的表达情况。结果HCC中her-2阳性表达率显著高于肝硬化及正常肝组织(χ2=6.482,P=0.032);HCC... 详细信息

目的研究her-2/neu在肝细胞癌(HCC)患者中的表达及其临床意义。方法应用免疫组织化学SP法检测30例HCC、10例肝硬化、10例正常肝组织中her-2蛋白的表达情况。结果HCC中her-2阳性表达率显著高于肝硬化及正常肝组织(χ2=6.482,P=0.032);HCC中her-2的表达与是否伴有肝硬化(P=0.041)、侵袭性(P=0.028)、Edmondson分级(P=0.012)有明显的相关性,与其他临床病理特征无明显相关性。her-2表达阳性患者的平均生存时间明显短于阴性表达患者(P=0.036)。结论her-2/neu基因在HCC的侵袭性生长转移和发展中可能具有一定的促进作用,在HCC组织中her-2/neu表达阳性的患者预示较差的预后。

关键词：肝细胞癌肝硬化 her-2/neu 免疫组织化学靶向治疗

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her-2/neu COX-2 PGE_2 P450arom在子宫内膜癌组织的表达及其相关性研究

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中国肿瘤临床 2011年第19期38卷 1183-1186,1195页

作者：刘丽学马晓欣贾玖丽李艳霞腾月中国医科大学附属盛京医院妇产科沈阳市110004

目的:探讨子宫内膜腺癌组织中her-2/neu、COX-2、PGE_2、P450arom4种生物学指标的表达,分析这4种生物学指标表达强度之间相关性及与临床病理因素的相关性。方法:应用逆转录-聚合酶链反应(RT-PCR)技术及酶联免疫吸附试验(ELISA)检测her-2... 详细信息

目的:探讨子宫内膜腺癌组织中her-2/neu、COX-2、PGE_2、P450arom4种生物学指标的表达,分析这4种生物学指标表达强度之间相关性及与临床病理因素的相关性。方法:应用逆转录-聚合酶链反应(RT-PCR)技术及酶联免疫吸附试验(ELISA)检测her-2/neu、COX-2、P450arom及PGE_2在正常子宫内膜、子宫内膜不典型增生和子宫内膜腺癌组织中的表达情况。结果:her-2/neu、COX-2、P450arom及PGE_2在癌组织中表达高于非癌组织,有显著性差异(Pher-2/neu与COX-2、P450arom表达强度呈正相关(r值分别为0.931,0.934,P2与P450arom表达强度亦呈正相关(r=0.908,P2mRNA和PGE_2表达强度变化呈正相关(r=0.552,Pher-2/neu、COX-2、P450arom及PGE_2可能存在协同作用,共同促进子宫内膜腺癌的发生发展。

关键词： her-2/neu COX-2 P450arom子宫内膜癌

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卵巢癌her-2/neu表达与肿瘤免疫细胞分布关系

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复旦学报（医学版） 2003年第5期30卷 426-429页

作者：朱芝玲李大金黄敏丽复旦大学附属妇产科医院上海200011

目的　研究上皮性卵巢癌细胞原癌基因her - 2 /neu表达与肿瘤微环境免疫活性细胞分布的关系 ,探讨过度表达原癌基因her - 2 /neu的上皮性卵巢癌临床分期晚、预后差的可能原因。了解过度表达her - 2 /neu基因的上皮性卵巢癌肿瘤微环境的... 详细信息

目的　研究上皮性卵巢癌细胞原癌基因her - 2 /neu表达与肿瘤微环境免疫活性细胞分布的关系 ,探讨过度表达原癌基因her - 2 /neu的上皮性卵巢癌临床分期晚、预后差的可能原因。了解过度表达her - 2 /neu基因的上皮性卵巢癌肿瘤微环境的免疫细胞分布。方法　用流式细胞仪检测上皮性卵巢癌细胞原癌基因her - 2 /neu的表达 ,同时用免疫组化方法检测肿瘤微环境中LCA+ 、CD+ 3 细胞分布情况。结果　 30 %卵巢癌细胞原癌基因her - 2 /neu呈过度表达 ,且卵巢癌细胞原癌基因her - 2 /neu表达与肿瘤微环境中LCA+ 、CD+ 3细胞分布呈负相关 ,前者r =- 0 .6 30 ,P =0 .0 0 0 1,后者r=- 0 .5 5 2 ,P =0 .0 0 0 9。结论　研究结果提示过度表达原癌基因her - 2 /neu的上皮性卵巢癌患者肿瘤微环境中浸润免疫细胞数减少 ,两者存在着一定的内在联系 ,可能发生肿瘤细胞的免疫逃逸。

关键词：卵巢癌 her-2/neu 基因表达肿瘤免疫细胞

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