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1.
To study the effect of interleukin-18 gene transfection on the tumorigenesis of breast cancer cell line Bacp37, human breast cancer cell line Bcap37 were transfected with Lipofectamine and selected by G418. The biological expression of rhIL-18 was tested by RT-PCR and ELISA method; nude mice were injected with Bcap37 cell with or without the hIL-18 gene. The hIL-18 cDNA was successfully integrated into Bcap37 cell; 126.3±4.5 pg hIL-18 secreted by one million transduced cells in 24 hours. Nude mice injected with IL-18 gene engineered Bcap37 cell had no tumor growth. These findings indicated that human breast cancer cells were successfully modified by the gene of IL-18 cytokine; the IL-18 gene engineered Bcap37 cells secreted hIL-18 and lost their tumorigenicity. The Bcap37 cells transduced with IL-18 gene may be used as breast cancer vaccine. 相似文献
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Enhancing cellular immune response to HBV M DNA vaccine in mice by codelivery of interleukin-18 recombinant 总被引：1，自引：0，他引：1
INTRODUCTIONChronicinfectionwithHBVaffectsmorethan250millionpeopleworldwide.Therearemorethan120millionchronicHBVcarriersinChina;appro-ximately10percentofthemremaininstateofchronichepatitisandhaveahighriskofdevelop-mentofcirrhosisandhepatocellularcarcinoma.ButthereisnoeffectivemethodtocontrolchronicHBVinfectionatpresent.Recentdataindicatedthatim-munotherapeuticstrategiesstimulatingbothcellularandhumoralimmuneresponsestoHBVantigensareessentialforcuringchronicHBVinfection(ChisariandFe… 相似文献
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Objective: To investigate the effect of interleukin-18 (IL-18) on immune response induced by plasmid encoding hepatitis B virus middle protein antigen and to explore new strategies for prophylactic and therapeutic HBV DNA vaccines. Methods: BALB/c mice were immunized with pCMV-M alone or co-immunized with pcDNA3-18 and pCMV-M and then their sera were collected for analysing anti-HBsAg antibody by ELISA; splenocytes were isolated for detecting specific CTL response and cytokine assay in vitro. Results: The anti-HBs antibody level of mice co-immunized with pcDNA3-18 and pCMV-M was slightly higher than that of mice immunized with pCMV-M alone, but there was not significantly different (P>0.05). Compared with mice injected with pCMV-M, the specific CTL cytotoxity activity of mice immunized with pcDNA3-18 and pCMV-M was significantly enhanced (P<0.05) and the level of IFN-γ in supernatant of splenocytes cultured with HBsAg in vitro was significantly elevated (P<0.05) while the level of IL-4 had no significant difference (P>0.05). Conclusion: The plasmid encoding IL-18 together with HBV M gene DNA vaccines may enhance specific TH1 cells and CTL cellular immune response induced in mice, so that IL-18 is a promising immune adjuvant. 相似文献
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川芎嗪对人乳腺癌MCF-7/ADM细胞多药耐药性的逆转及P-糖蛋白表达的影响 总被引：2，自引：0，他引：2
[目的]探讨川芎嗪(TMP)对人乳腺癌MCF-7/ADM细胞的耐药逆转及其对该细胞P-糖蛋白(P-gp)表达的影响.[方法]MTT法测定细胞的药敏性和抗药性逆转,流式细胞术检测耐药细胞P-糖蛋白的表达.[结果]非细胞毒性剂量(320μg/m l)和低毒剂量(1 250μg/m l)的川芎嗪均能显著降低MCF-7/ADM细胞的IC50(P<0.01),逆转倍数分别为2.14和2.80倍;并使该细胞P-gp的表达率由(90.60±0.40)%分别降低至(69.10±1.65)%和(60.30±1.25)%.[结论]川芎嗪可部分逆转人乳腺癌MCF-7/ADM细胞对阿霉素的耐药性,其逆转机制与抑制该细胞P-gp的表达有关. 相似文献
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This paper aims to screen and identify sphere clone cells with characteristics similar to cancer stem cells in human gallbladder
cancer cell line GBC-SD. GBC-SD cells were cultured in a serum-free culture medium with different concentrations of the chemotherapeutic
drug cisplatin for generating sphere clones. The mRNA expressions of stem cell-related genes CD133, OCT-4, Nanog, and drug
resistance genes ABCG2 and MDR-1 in sphere clones were detected by quantitative real-time polymerase chain reaction (PCR).
Stem cell markers were also analyzed by flow cytometry and immunofluorescent staining. Different amounts of sphere clones
were injected into nude mice to test their abilities to form tumors. Sphere clones were formed in serum-free culture medium
containing cisplatin (30 μmol/L). Flow cytometry results demonstrated that the sphere clones expressed high levels of stem
cell markers CD133+ (97.6%) and CD44+ (77.9%) and low levels of CD24+ (2.3%). These clones also overexpressed the drug resistance genes ABCG2 and MDR-1. Quantitative real-time PCR showed that
sphere clones expressed stem cell genes Nanog and OCT-4 284 and 266 times, respectively, more than those in the original GBC-SD
cells. Immunofluorescent staining showed that sphere clones overexpressed OCT-4, Nanog, and SOX-2, and low expressed MUC1
and vimentin. Tumor formation experiments showed that 1×103 sphere clone cells could induce much larger tumors in nude mice than 1×105 GBC-SD cells. In conclusion, sphere clones of gallbladder cancer with stem cell-like characteristics can be obtained using
suspension cultures of GBC-SD cells in serum-free culture medium containing cisplatin. 相似文献
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目的探讨银杏叶总黄酮对体外培养的人肝癌细胞HepG2增殖与凋亡的影响。方法将银杏叶总黄酮作用于体外培养的人肝癌细胞HepG2,MTT法检测其对HepG2细胞增殖的影响,缺口末端核苷标记（TUNNEL）法检测其对HepG2细胞凋亡的影响。结果银杏叶总黄酮对体外培养的人肝癌细胞HepG2的增殖效率下降,使凋亡细胞数增加（P〈0．01）,且呈剂量依赖效应。结论银杏叶总黄酮对体外培养的人HepG2细胞增殖有抑制作用,并能诱导细胞凋亡。 相似文献
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Jia-wei Zhang Valentina Rubio Shu Zheng Zheng-zheng Shi 《Journal of Zhejiang University. Science. B》2009,10(11):796-804
To explore the role of a novel Obg-like ATPase 1 (OLA1) in cancer metastasis, small interference RNA (siRNA) was used to knockdown the protein, and the cells were subjected to in vitro cell migration and invasion assays. Knockdown of OLA1 significantly inhibited cell migration and invasion in breast cancer cell line MDA-MB-231. The knockdown caused no changes in cell growth but affected ROS production. In wound-healing assays, decreased ROS in OLA1-knockdown cells were in situ asso-ciated with the cells' decreased motile morphology. Further, treatment of N-acetylcysteine, a general ROS scavenger, blunted the motility and invasiveness of MDA-MB-231 cells, similar to the effect of OLA1-knockdown. These results suggest that knock-down of OLA1 inhibits breast cancer cell migration and invasion through a mechanism that involves the modulation of intracel-lular ROS levels. 相似文献
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目的探讨银杏叶黄酮单体成分槲皮素体外对人肝癌细胞HepG2增殖与凋亡的影响。方法将银杏叶黄酮单体成分槲皮素作用于体外培养的人肝癌细胞HepG2,MTT法检测其对HepG2细胞增殖的影响,缺口末端核苷标记（TUNNEL）法检测其对HepG2细胞凋亡的影响。结果银杏叶黄酮单体成分槲皮素使体外培养的人肝癌细胞HepG2的增殖效率下降,使凋亡细胞数增加（P〈0.01）,两者均呈剂量依赖效应。结论银杏叶黄酮单体成分槲皮素具有与银杏叶总黄酮相似的作用,对体外培养的人HepG2细胞增殖有抑制作用,并能诱导细胞凋亡。 相似文献
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白细胞介素18及其生物活性 总被引：2，自引：0，他引：2
白细胞介素18(IL-18)是新近发现的一种细胞因子，能诱导TH1细胞等产生IFN-r，GM-CSF等细胞因子及Fas配体，具有增强THI细胞和NK细胞的细胞毒作用等免疫调节功能，在抗病原微生物感染、抗肿瘤及抗超敏反映等方面具有潜在的应用前景，同时，它也是内毒素诱导的小鼠肝坏死性休克的关键因子。 相似文献
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Qiong-yan Lin Yi-feng Wang Hui-nan Weng Xiu-jie Sheng Qing-ping Jiang Zhi-ying Yang 《Journal of Zhejiang University. Science. B》2012,13(11):894-903
Background and objective: Gonadotropin-releasing hormone (GnRH) plays an important role in the regulation of ovarian function and ovarian cancer cell growth. In this study, we determined whether administration of the GnRH agonist (GnRHa), triporelin, prior to cisplatin treatment affects cisplatin and/or prevents cisplatin-induced ovarian damage. Methods: nu/nu mice were injected with ovarian cancer OVCAR-3 cells intraperitoneally. After two weeks, the mice were treated with saline (control), cisplatin, GnRHa, or cisplatin plus GnRHa for four weeks. At the end of the experimental protocol, blood, tumor, ovary, and uterine tissues were resected for hematoxylin and eosin (H&E) staining, immunohistochemical analyses of Ki67, nuclear factor-κB (NF-κB), and caspase-3, transmission electron microscopy of apoptosis, or enzyme-linked immunosorbent assay (ELISA) analyses of anti-Mullerian hormone (AMH). Results: Cisplatin treatment effectively inhibited tumor growth in mice treated with human ovarian cancer cells; however the treatment also induced considerable toxicity. Immunohistochemical analyses showed that Ki67 expression was reduced in cisplatin-treated mice compared to control (P<0.05), but there was no statistically significant differences between cisplatin-treated mice and cisplatin plus GnRHa-treated mice (P>0.05), while expressions of NF-κB and caspase-3 were reduced and induced, respectively, in cisplatin-treated mice and cisplatin plus GnRHa-treated mice. Apoptosis occurred in the GnRHa, cisplatin, and cisplatin plus GnRHa-treated mice, but not in control mice. Ovaries exposed to GnRHa in both GnRHa mice and cisplatin-treated mice (combination group) had significantly more primordial and growth follicles and serum levels of AMH than those in the control mice and cisplatin-treated mice (P<0.05). Conclusions: Administration of GnRHa to mice significantly decreased the extent of ovarian damage induced by cisplatin, but did not affect the anti-tumor activity of cisplatin. 相似文献
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Sun Jie Huang He Zhu Yuan-yuan Lan Jian-ping Li Jing-yuan Lai Xiao-yu Yu Jian 《Journal of Zhejiang University. Science. B》2005,6(12):1141-1147
Objective: Detecting the expression and mutation of human telomeric repeat binding factor (hTRF1) in 10 malignant hematopoietic cell line cells on the base of determining its genomic structure and its four pseudogenes
to clarify if hTRF1 mutation is one of the factors of the activation of telomerase. Methods: hTRF1cDNA sequences were obtained from GenBank, its genome structure and pseudogenes were forecasted by BLAST and other biology
information programs and then testified by sequencing. Real-time RT-PCR was used to detect the expression of hTRF1mRNA in 10 cell line cells, including myelogenous leukemia cell lines K562, HL-60, U-937, NB4, THP-1, HEL and Dami; lymphoblastic
leukemia cell lines 6T-CEM, Jurkat and Raji. Telomerase activities of cells were detected by using telomeric repeat amplification
(TRAP)-ELISA protocol. PCR and sequencing were used to detect mutation of each exon of hTRF1 in 10 cell line cells. Results: hTRF1 gene, mapped to 8q13, was divided into 10 exons and spans 38.6 kb. Four processed pseudogenes of hTRF1 located on chromosome 13, 18, 21 and X respectively, was named as ψhTRF1-13, ψhTRF1-18, ψhTRF1-21 and ψhTRF1-X respectively. All cell line cells showed positive telomerase activity. The expression of hTRF1 was significantly lower in malignant hematopoietic cell lines cells (0.0338, 0.0108:_0.0749) than in normal mononuclear cells
(0.0493, 0.0369:_0.128) (P=0.004). But no significant mutation was found in all exons of hTRF1 in 10 cell line cells. Four variants were found in part of intron 1, 2 and 8 of hTRF1. Their infection on gene function is unknown and needs further studies. Conclusion: hTRF1 mutation is probably not one of the main factors for telomerase activation in malignant hematopoietic disease.
Project supported by the National Basic Research Program (973) of China (No. 2002CB713700) and the National Natural Science
Foundation of China (No. 39870339) 相似文献
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Two novel sugar-conjugated 5-fluorocytosine (5-FC) antineoplastic compounds were designed and synthesized to improve the selective drug uptake by targeting the tumor-specific glucose transporter (GLUT).The antitumor activity of these compounds was evaluated in four different human cancer cell lines:A549 (human lung cancer cell line),HT29 (human colorectal cancer cell line),H460 (human lung cancer cell line),and PC3 (human prostate cancer cell line).The sugar conjugates exhibited cytotoxicity similar to or higher than 5-FC and 1-hexylcarbamoyl-5-FC in A549,HT29,H460,and PC3.Furthermore,GLUT-mediated transport of the glycoconjugate was investigated with GLUT inhibitor-mediated cytotoxicity analysis in a GLUT-overexpressing HT29 cell line.The cell-killing potency of 5-FC glycoconjugate was found to depend significantly on the GLUT inhibitor,and the cellular uptake of molecules was regulated by GLUT-mediated transport.All the results demonstrate the potential advantages of glycoconjugation for Warburg effect-targeted drug design. 相似文献
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Objective: To investigate the infection of human embryo fibroblast cell line HF cells by CMV as well as the effects of CMV
on β-actin mRNA and microfilaments. Methods: HF cells shape was observed after the infection of CMV. RT-PCR assay was used
to detect the mRNA expression of CMV immediate early (IE) gene, β-actin and GAPDH genes of HF cells infected by CMV. CMV particles
and cell microfilaments were detected with electron microscope. Results: Shape of HF cell changed after the infection by CMV.
HF cells infected by CMV could express IE mRNA and the expression of β-actin mRNA decreased in a time-and titer-dependent
manner compared with the uninfected HF cells whose expression of GAPDH mRNA did not change much. CMV particles were found
with electron microscope in the cells. Microfilaments were ruptured and shortened after the infection of CMV. Conclusion:
CMV can not only infect human embryo fibroblast cells line HF cells and replicate in the cells, but can also affect the expression
of β-actin mRNA and the microfilaments.
Project (No. 001103058) supported by the Key Program of Science and Technology Bureau of Zhejiang Province, China 相似文献
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目的：通过对乳腺癌原代细胞培养。检测化疗药物对癌细胞的抑制率。探讨乳腺癌不同组织学类型、年龄及淋巴结转移情况对化疗药物的敏感性。方法：收集109例手术切除新鲜乳腺癌标本．进行原代细胞培养。采用四甲基偶氮唑盐（MTT）法检测癌细胞对三组抗癌药物的敏感性。结果：乳腺癌的敏感率由高到低依次为CAF（33．0％）、TA（31．2％）和CMF（26．6％）。无论年龄、组织学类型及是否有淋巴结转移各组间比较其敏感性均无显著差异（P〉0．05）。结论：1．MTT法对乳腺癌治疗时选择化疗药物具有重要价值；2．乳腺癌细胞对CAF、TA、CMF敏感。 相似文献
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IMP3（胰岛素生长因子Ⅱ的mRNA的结合蛋白3）是一个新发现的癌胚胎的RNA结合蛋白,研究发现其参与细胞生长和细胞迁移在胚胎发育的早期阶段．该研究利用实时定量逆转录PCR对IMP3在乳腺癌的诊断和治疗中的应用潜力进行了研究和评估．检测了不同类型乳腺癌样本中IMP3的表达情况,定量PCR结果分析中应用2也小。方法进行了归一化比较．研究结果表明,IMP3基因表达与乳腺癌病人肿瘤的大小及淋巴转移存在明显的相关性（P〈0．01）．不同的临床阶段和病理分期与IMP3的表达水平之间也存在一定的相关性,表明差异不显著（P〉0．05）．该研究表明IMP3作为一种新的肿瘤相关因子,其表达水平与乳腺癌也存在一定的相关性,显示了其作为一种潜在的肿瘤标志物的可能性． 相似文献
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目的从昆明鼠睾丸中克隆Bmi1基因,构建真核表达载体,并转染支持细胞,以便用作培养精原干细胞（SSCs）的滋养层.方法以5日龄昆明鼠为材料,提取小鼠睾丸组织中总RNA后,以RT-PCR技术克隆小鼠睾丸Bmi1基因,构建真核表达载体,并转染TM4细胞（睾丸支持细胞株）,在转染后40 h进行免疫荧光鉴定.结果成功克隆小鼠睾丸Bmi1基因的cDNA,测序正确;免疫荧光细胞染色显示,转染后的支持细胞中有Bmi1蛋白表达.结论本研究为以转染了Bmi1基因的支持细胞作饲养层培养SSCs奠定了基础. 相似文献