目的研究非小细胞肺癌患者血清中卵泡抑素(FS)和粘蛋白(MUC1)表达与肿瘤的发生发展与转移的关系及其临床意义。方法采集60例肺部肿瘤入院患者的血清样本,其中非小细胞肺癌40例,肺部良性肿瘤20例,分别采用ELISA法和Can Ag MUC1检测试剂...目的研究非小细胞肺癌患者血清中卵泡抑素(FS)和粘蛋白(MUC1)表达与肿瘤的发生发展与转移的关系及其临床意义。方法采集60例肺部肿瘤入院患者的血清样本,其中非小细胞肺癌40例,肺部良性肿瘤20例,分别采用ELISA法和Can Ag MUC1检测试剂盒检测其血清中FS和MUC1含量。结果 40例非小细胞肺癌患者血清中FS和MUC1的血清含量、阳性率均高于对照组(P<0.01)。结论 FS、MUC1与肿瘤发生、发展、生物学特征以及转移密切相关,联和检测这些指标有利于非小细胞肺癌的早期诊断以及预后判断。展开更多
Two eukaryotic vectors expressing 9 tandem repeats of human MUCI（VNTR）, VR1012-VNTR, and pEGFP-VNTR, were constructed by cloning VNTR gene into VR1012 and pEGFP, respectively. VNTR stably expressing murine Lewis lun...Two eukaryotic vectors expressing 9 tandem repeats of human MUCI（VNTR）, VR1012-VNTR, and pEGFP-VNTR, were constructed by cloning VNTR gene into VR1012 and pEGFP, respectively. VNTR stably expressing murine Lewis lung carcinoma（LLC） cell line（VNTR^＋ LLC） was established by Lipofectamine-mediated transfection of pEGFP-VNTR into LLC cells. The EGFP expression was observed under a fluorescent microscope and VNTR expression in VNTR^＋ LLC cells was confirmed by means of Western blotting. A syngenic graft tumor model was generated by subcutaneous injection of VNTR^＋ LLC cells into C57/BL6 mice and tumor size increased rapidly with time and in a cell qumber dependent manner. VNTR mRNA expression in the tumor formed was confirmed by RT-PCR. After the third immunization mice were challenged subcutaneously with 5×10^5 VNTR^＋ LLC cells, a significant reduction of subcutaneous tumor growth was observed in the groups immunized with VNTR plasmid DNA compared with that in the groups immunized with the vector DNA alone. Thus, the suppression of subcutaneous tumor was antigen-specific. This model is useful for the development of tumor vaccines targeting MUCI VNTRs.展开更多
目的获得具有较好致瘤性、在动物体内保留表达外源基因MUC1 VNTR(mucin1,variable number tandemrepeats)能力的稳定转染细胞株B16-GFP-MUC1,供MUC1靶向药物研究中体内抑瘤实验所用到的肿瘤模型。方法用脂质体转染的方法和有限稀释法获...目的获得具有较好致瘤性、在动物体内保留表达外源基因MUC1 VNTR(mucin1,variable number tandemrepeats)能力的稳定转染细胞株B16-GFP-MUC1,供MUC1靶向药物研究中体内抑瘤实验所用到的肿瘤模型。方法用脂质体转染的方法和有限稀释法获得单克隆细胞株;将该细胞株进行体内传代后,流式分选GFP阳性细胞群,并对分选后细胞用有限稀释法进行单克隆化。用流式细胞仪和共聚焦荧光显微镜观察各时期细胞克隆的GFP表达情况。结果脂质体转染方法得到的在G418压力选择下存在的细胞B16-1,其GFP阳性细胞比率最高为80%左右;致瘤性较差,75d内仅87.5%小鼠出瘤;体外的稳定性观察中,在无抗生素的培养液中观察5周,其阳性率降至20%左右;经过体内接种后仅有10%左右的肿瘤细胞保留了外源基因的表达。而经过体内传代和流式分选后得到的单克隆细胞群B16-39,其GFP阳性细胞比率>90%;致瘤性为20d内100%小鼠出瘤;体外培养8周,GFP阳性的细胞比率>90%;体内剥离后的肿瘤细胞的GFP阳性细胞比率均>90%。结论对于用于体内动物模型的稳定转染细胞,通过体内传代,流式分选并单克隆化,是获得致瘤性好,体内体外较为稳定的稳定转染细胞株的一个可行方案。展开更多
Background and Objective Lymph node, peripheral blood and bone marrow from NSCLC patients have undetectable micro-metastasis by general method, and the tumor micrometastasis
文摘目的研究非小细胞肺癌患者血清中卵泡抑素(FS)和粘蛋白(MUC1)表达与肿瘤的发生发展与转移的关系及其临床意义。方法采集60例肺部肿瘤入院患者的血清样本,其中非小细胞肺癌40例,肺部良性肿瘤20例,分别采用ELISA法和Can Ag MUC1检测试剂盒检测其血清中FS和MUC1含量。结果 40例非小细胞肺癌患者血清中FS和MUC1的血清含量、阳性率均高于对照组(P<0.01)。结论 FS、MUC1与肿瘤发生、发展、生物学特征以及转移密切相关,联和检测这些指标有利于非小细胞肺癌的早期诊断以及预后判断。
文摘Two eukaryotic vectors expressing 9 tandem repeats of human MUCI（VNTR）, VR1012-VNTR, and pEGFP-VNTR, were constructed by cloning VNTR gene into VR1012 and pEGFP, respectively. VNTR stably expressing murine Lewis lung carcinoma（LLC） cell line（VNTR^＋ LLC） was established by Lipofectamine-mediated transfection of pEGFP-VNTR into LLC cells. The EGFP expression was observed under a fluorescent microscope and VNTR expression in VNTR^＋ LLC cells was confirmed by means of Western blotting. A syngenic graft tumor model was generated by subcutaneous injection of VNTR^＋ LLC cells into C57/BL6 mice and tumor size increased rapidly with time and in a cell qumber dependent manner. VNTR mRNA expression in the tumor formed was confirmed by RT-PCR. After the third immunization mice were challenged subcutaneously with 5×10^5 VNTR^＋ LLC cells, a significant reduction of subcutaneous tumor growth was observed in the groups immunized with VNTR plasmid DNA compared with that in the groups immunized with the vector DNA alone. Thus, the suppression of subcutaneous tumor was antigen-specific. This model is useful for the development of tumor vaccines targeting MUCI VNTRs.
文摘目的获得具有较好致瘤性、在动物体内保留表达外源基因MUC1 VNTR(mucin1,variable number tandemrepeats)能力的稳定转染细胞株B16-GFP-MUC1,供MUC1靶向药物研究中体内抑瘤实验所用到的肿瘤模型。方法用脂质体转染的方法和有限稀释法获得单克隆细胞株;将该细胞株进行体内传代后,流式分选GFP阳性细胞群,并对分选后细胞用有限稀释法进行单克隆化。用流式细胞仪和共聚焦荧光显微镜观察各时期细胞克隆的GFP表达情况。结果脂质体转染方法得到的在G418压力选择下存在的细胞B16-1,其GFP阳性细胞比率最高为80%左右;致瘤性较差,75d内仅87.5%小鼠出瘤;体外的稳定性观察中,在无抗生素的培养液中观察5周,其阳性率降至20%左右;经过体内接种后仅有10%左右的肿瘤细胞保留了外源基因的表达。而经过体内传代和流式分选后得到的单克隆细胞群B16-39,其GFP阳性细胞比率>90%;致瘤性为20d内100%小鼠出瘤;体外培养8周,GFP阳性的细胞比率>90%;体内剥离后的肿瘤细胞的GFP阳性细胞比率均>90%。结论对于用于体内动物模型的稳定转染细胞,通过体内传代,流式分选并单克隆化,是获得致瘤性好,体内体外较为稳定的稳定转染细胞株的一个可行方案。
基金supported by the grants from the Key Project of National Natural Science Foundation of China (No.30430300 , to Qinghua ZHOU)Key Projects of Tianjin Sci-Tech Support Program (No. 07SYSYSF05000 and No. 06YF-SZSF05300, to Qinghua ZHOU)
文摘Background and Objective Lymph node, peripheral blood and bone marrow from NSCLC patients have undetectable micro-metastasis by general method, and the tumor micrometastasis
