Objective： HOXB7 gene is a kind of transcription regulator over-expressed in malignant melanoma （MM） cell lines. It can specifically up-regulate the expression of angiogenic factors and tumor growth factors such as...Objective： HOXB7 gene is a kind of transcription regulator over-expressed in malignant melanoma （MM） cell lines. It can specifically up-regulate the expression of angiogenic factors and tumor growth factors such as bFGF, GROa, VEGF and induce angiogenesis in melanoma, resulting in the proliferation and metastasis of tumor cells. We designed and synthesized HOXB7 specific siRNA to study its interfering effect on the expressions of HOXB7 and bFGF genes in melanoma A375 cell line and the biologic characteristics of A375 cells. Methods： Three synthesized siRNA with different sequences were separately transfected into A375 cells by lipofecter 2000. The expression of HOXB7 and bFGF mRNA in transfected cells was detected by RT-PCR 24 and 48 hours after transduction. The expression of bFGF protein in the transfected cells were detected by flowcytometry 48 hours after transfection. MTT assay was used to analyze the cell proliferation rate of siRNA transfected cells. Based on the in vitro experiment results, one effective siRNA sequence was selected for the construction of in vivo siRNA expression vector. Then, a malignant melanoma animal model was established. The siRNA expression plasmid was injected into the tumor foci and its influence on the growth and angiogenesis of tumor was observed. Results： The mRNA expressions of both HOXB7 and bFGF genes in the A375 cells reduced significantly 24 and 48 hour after transfection of siRNA. Expression level of the protein of angiogenic factor bFGF induced by HOXB7 gene in siRNA transfected cells was significantly lower than that in control cells 48 hours after transduction. Cell proliferation was also suppressed in siRNA transfected cells. Two of the three siRNA strands showed prominent interference effect. The in vivo study indicated that the tumor size and the microvessel density in the tumor both reduced after injection of HOXB7siRNA plasmid. Conclusion： Down-regulation of HOXB7 gene expression can effectively reduce the expression of angiogenic factor bFGF and the proliferation of MM cells. Besides, the growth and angiogenesis of MM tumor were also inhibited.展开更多
分子伴侣热休克蛋白90(heat shock protein 90,HSP90)能够通过泛素化途径保护细胞内蛋白质功能,其在肿瘤细胞中呈过表达状态,维持肿瘤细胞生长、增殖、抗凋亡及转移能力。Ganetespib是目前广泛应用于多种肿瘤治疗临床试验的小分子HSP90...分子伴侣热休克蛋白90(heat shock protein 90,HSP90)能够通过泛素化途径保护细胞内蛋白质功能,其在肿瘤细胞中呈过表达状态,维持肿瘤细胞生长、增殖、抗凋亡及转移能力。Ganetespib是目前广泛应用于多种肿瘤治疗临床试验的小分子HSP90抑制剂,其单药具有高效能的抗肿瘤活性,联合用药能增强标准化疗或其他靶向治疗疗效,且能同时克服多种肿瘤的耐药机制,本文就Ganetespib在多种人类恶性实体肿瘤治疗中的疗效进行讨论。展开更多
目的:分析非小细胞肺癌(NSCLC)中黑素瘤相关抗原D4(MAGE-D4)基因启动子甲基化程度与MAGE-D4 mRNA表达水平及患者临床病理特征的关系。方法:采用甲基化特异性PCR(MSP)法检测38例NSCLC患者癌组织及其癌旁组织中MAGE-D4启动子甲基化程度,...目的:分析非小细胞肺癌(NSCLC)中黑素瘤相关抗原D4(MAGE-D4)基因启动子甲基化程度与MAGE-D4 mRNA表达水平及患者临床病理特征的关系。方法:采用甲基化特异性PCR(MSP)法检测38例NSCLC患者癌组织及其癌旁组织中MAGE-D4启动子甲基化程度,实时荧光定量PCR(qPCR)法检测组织中MAGE-D4 m RNA相对表达量,分析MAGE-D4启动子甲基化频率与患者临床病理特征的关系。结果:肺癌组织中MAGE-D4基因甲基化频率为23.68%,明显低于癌旁组织的86.84%(P<0.05);而癌组织中MAGE-D4 m RNA表达水平则显著高于癌旁组织(5.49±5.65 vs.1.44±1.08,P<0.05),MAGE-D4表达量与启动子甲期化程度呈负相关关系(r=-0.663,P<0.05)。肿瘤直径>3 cm的肺癌患者MAGE-D4甲基化发生率明显低于肿瘤直径≤3 cm患者(P<0.05)。结论:NSCLC中MAGE-D4基因转录活性的增强与其启动子低甲基化相关;肿瘤直径较大的NSCLC患者MAGE-D4甲基化发生率较低。展开更多
基金the grants from the Research Foundation of Science & Technology Bureau of Guangzhou(2004Z2-E0011)the Guangdong Province Natural Science Foundation(5002318)
文摘Objective： HOXB7 gene is a kind of transcription regulator over-expressed in malignant melanoma （MM） cell lines. It can specifically up-regulate the expression of angiogenic factors and tumor growth factors such as bFGF, GROa, VEGF and induce angiogenesis in melanoma, resulting in the proliferation and metastasis of tumor cells. We designed and synthesized HOXB7 specific siRNA to study its interfering effect on the expressions of HOXB7 and bFGF genes in melanoma A375 cell line and the biologic characteristics of A375 cells. Methods： Three synthesized siRNA with different sequences were separately transfected into A375 cells by lipofecter 2000. The expression of HOXB7 and bFGF mRNA in transfected cells was detected by RT-PCR 24 and 48 hours after transduction. The expression of bFGF protein in the transfected cells were detected by flowcytometry 48 hours after transfection. MTT assay was used to analyze the cell proliferation rate of siRNA transfected cells. Based on the in vitro experiment results, one effective siRNA sequence was selected for the construction of in vivo siRNA expression vector. Then, a malignant melanoma animal model was established. The siRNA expression plasmid was injected into the tumor foci and its influence on the growth and angiogenesis of tumor was observed. Results： The mRNA expressions of both HOXB7 and bFGF genes in the A375 cells reduced significantly 24 and 48 hour after transfection of siRNA. Expression level of the protein of angiogenic factor bFGF induced by HOXB7 gene in siRNA transfected cells was significantly lower than that in control cells 48 hours after transduction. Cell proliferation was also suppressed in siRNA transfected cells. Two of the three siRNA strands showed prominent interference effect. The in vivo study indicated that the tumor size and the microvessel density in the tumor both reduced after injection of HOXB7siRNA plasmid. Conclusion： Down-regulation of HOXB7 gene expression can effectively reduce the expression of angiogenic factor bFGF and the proliferation of MM cells. Besides, the growth and angiogenesis of MM tumor were also inhibited.
文摘分子伴侣热休克蛋白90(heat shock protein 90,HSP90)能够通过泛素化途径保护细胞内蛋白质功能,其在肿瘤细胞中呈过表达状态,维持肿瘤细胞生长、增殖、抗凋亡及转移能力。Ganetespib是目前广泛应用于多种肿瘤治疗临床试验的小分子HSP90抑制剂,其单药具有高效能的抗肿瘤活性,联合用药能增强标准化疗或其他靶向治疗疗效,且能同时克服多种肿瘤的耐药机制,本文就Ganetespib在多种人类恶性实体肿瘤治疗中的疗效进行讨论。
文摘目的:分析非小细胞肺癌(NSCLC)中黑素瘤相关抗原D4(MAGE-D4)基因启动子甲基化程度与MAGE-D4 mRNA表达水平及患者临床病理特征的关系。方法:采用甲基化特异性PCR(MSP)法检测38例NSCLC患者癌组织及其癌旁组织中MAGE-D4启动子甲基化程度,实时荧光定量PCR(qPCR)法检测组织中MAGE-D4 m RNA相对表达量,分析MAGE-D4启动子甲基化频率与患者临床病理特征的关系。结果:肺癌组织中MAGE-D4基因甲基化频率为23.68%,明显低于癌旁组织的86.84%(P<0.05);而癌组织中MAGE-D4 m RNA表达水平则显著高于癌旁组织(5.49±5.65 vs.1.44±1.08,P<0.05),MAGE-D4表达量与启动子甲期化程度呈负相关关系(r=-0.663,P<0.05)。肿瘤直径>3 cm的肺癌患者MAGE-D4甲基化发生率明显低于肿瘤直径≤3 cm患者(P<0.05)。结论:NSCLC中MAGE-D4基因转录活性的增强与其启动子低甲基化相关;肿瘤直径较大的NSCLC患者MAGE-D4甲基化发生率较低。
