目的:探讨Bcl-3在乳腺癌中的表达及沉默后对乳腺癌细胞生物学行为的影响。方法:选取80例乳腺癌组织及相应癌旁组织,采用免疫组织化学法检测其中Bcl-3蛋白表达,并分析癌组织中Bcl-3表达与临床病理参数间的关系;将乳腺癌MDA-MB-231细胞随...目的:探讨Bcl-3在乳腺癌中的表达及沉默后对乳腺癌细胞生物学行为的影响。方法:选取80例乳腺癌组织及相应癌旁组织,采用免疫组织化学法检测其中Bcl-3蛋白表达,并分析癌组织中Bcl-3表达与临床病理参数间的关系;将乳腺癌MDA-MB-231细胞随机分为空白对照组(常规培养),阴性对照组(转染对照si RNA)和Bcl-3 si RNA组(转染Bcl-3 si RNA)。转染48 h后,免疫印迹法检测各组Bcl-3表达;MTT、平板克隆实验及流式细胞术分别检测各组细胞增殖能力、克隆形成能力和细胞凋亡。结果:Bcl-3蛋白在乳腺癌及相应癌旁组织中阳性表达率分别为72.5%和51.3%。乳腺癌组织中Bcl-3蛋白表达与组织学分级、淋巴结转移及Her-2表达呈正相关(P<0.05),而与年龄、肿瘤大小、病理分型及雌、孕激素受体无关(P>0.05)。空白对照组和阴性对照组Bcl-3表达水平明显高于Bcl-3 si RNA组(P均<0.01)。与空白对照组和阴性对照组比较,Bcl-3 si RNA组细胞增殖率和克隆形成率降低,细胞凋亡率增高(P均<0.01)。结论:Bcl-3在乳腺癌组织中呈高表达,其沉默后起抑癌作用,乳腺癌细胞增殖能力及克隆形成率降低、凋亡率增高。展开更多
目的化疗耐药是乳腺癌复发转移、治疗效果不理想的主要因素。本文探讨miR-142-3p通过靶向高迁移率族蛋白1(high-mobility group box 1,HMGB1)提高乳腺癌化疗敏感性的分子机制。方法实时荧光定量PCR检测人乳腺癌MCF-7细胞及阿霉素耐药株M...目的化疗耐药是乳腺癌复发转移、治疗效果不理想的主要因素。本文探讨miR-142-3p通过靶向高迁移率族蛋白1(high-mobility group box 1,HMGB1)提高乳腺癌化疗敏感性的分子机制。方法实时荧光定量PCR检测人乳腺癌MCF-7细胞及阿霉素耐药株MCF-7/DOX细胞中的miR-142-3p水平;MTT法检测阿霉素(doxorubicin,DOX)处理后各组的增殖情况、流式细胞术检测凋亡率、Western blot检测HMGB1和自噬有关蛋白的表达水平;双荧光素酶报告实验评价miR-142-3p对HMGB1的靶向调控作用。结果阿霉素耐药株MCF-7/DOX细胞中的miR-142-3p水平明显下调。miR142-3p的过度表达增强了乳腺癌细胞对阿霉素的敏感性和提高阿霉素诱导的凋亡比率。HMGB1是乳腺癌细胞中miR-142-3p的直接功能靶点,HMGB1的过表达可以明显解除由miR-142-3p上调所产生的细胞凋亡和自噬抑制。结论miR-142-3p的过表达可能通过抑制自噬靶向HMGB1,增强乳腺癌细胞对阿霉素的化学敏感性。miR-142-3p/HMGB1为提高乳腺癌对药物的敏感性提供了新的靶点,具有一定价值。展开更多
Our previous studies indicate that phosphatidylinositol 4-kinase Ila can promote the growth of multi-malignant tumors via HER-2/PI3K and MAPK pathways. However, the molecular mechanisms of this pathway and its potenti...Our previous studies indicate that phosphatidylinositol 4-kinase Ila can promote the growth of multi-malignant tumors via HER-2/PI3K and MAPK pathways. However, the molecular mechanisms of this pathway and its potential for clinical application remain unknown. In this study, we found that PI4KIla could be an ideal combinatorial target for EGFR treatment via regulating EGFR degradation. Results showed that PI4KIla knockdown reduced EGFR protein level, and the expression of Pi4KIla shows a strong correlation with EGFR in human breast cancer tissues （r= 0.77, P 〈 0.01）. PI4KIla knockdown greatly prolonged the effects and decreased the effective dosage of AG-1478, a specific inhibitor of EGFR. In addition, it significantly enhanced AG1478-induced inhibition of tumor cell survival and strengthened the effect of the EGFR-targeting anti-cancer drug Iressa in xenograft tumor models. Mechanistically, we found that PI4KIla suppression increased EGFR ligand-independent degradation. Quantitative proteomic analysis by stable isotope labeling with amino acids in cell culture （SlLAC） and LC-MS/MS suggested that HsPg0 mediated the effect of PI4KIla on EGFR. Furthermore, we found that combined inhibition of PI4KIla and EGFR suppressed both PI3K/AKT and MAPK/ERK pathways, and resulted in downregulation of multiple oncogenes like PRDX2, FASN, MTA2, ultimately leading to suppression of tumor growth. Therefore, we conclude that combined inhibition of PI4KIla and EGFR exerts a multiple anti-tumor effect. Dual inhibition of EGFR at protein and activity level via combinatorial blocking of PI4KIla presents a novel strategy to combat EGFR-dependent tumors.展开更多
文摘目的:探讨Bcl-3在乳腺癌中的表达及沉默后对乳腺癌细胞生物学行为的影响。方法:选取80例乳腺癌组织及相应癌旁组织,采用免疫组织化学法检测其中Bcl-3蛋白表达,并分析癌组织中Bcl-3表达与临床病理参数间的关系;将乳腺癌MDA-MB-231细胞随机分为空白对照组(常规培养),阴性对照组(转染对照si RNA)和Bcl-3 si RNA组(转染Bcl-3 si RNA)。转染48 h后,免疫印迹法检测各组Bcl-3表达;MTT、平板克隆实验及流式细胞术分别检测各组细胞增殖能力、克隆形成能力和细胞凋亡。结果:Bcl-3蛋白在乳腺癌及相应癌旁组织中阳性表达率分别为72.5%和51.3%。乳腺癌组织中Bcl-3蛋白表达与组织学分级、淋巴结转移及Her-2表达呈正相关(P<0.05),而与年龄、肿瘤大小、病理分型及雌、孕激素受体无关(P>0.05)。空白对照组和阴性对照组Bcl-3表达水平明显高于Bcl-3 si RNA组(P均<0.01)。与空白对照组和阴性对照组比较,Bcl-3 si RNA组细胞增殖率和克隆形成率降低,细胞凋亡率增高(P均<0.01)。结论:Bcl-3在乳腺癌组织中呈高表达,其沉默后起抑癌作用,乳腺癌细胞增殖能力及克隆形成率降低、凋亡率增高。
文摘目的化疗耐药是乳腺癌复发转移、治疗效果不理想的主要因素。本文探讨miR-142-3p通过靶向高迁移率族蛋白1(high-mobility group box 1,HMGB1)提高乳腺癌化疗敏感性的分子机制。方法实时荧光定量PCR检测人乳腺癌MCF-7细胞及阿霉素耐药株MCF-7/DOX细胞中的miR-142-3p水平;MTT法检测阿霉素(doxorubicin,DOX)处理后各组的增殖情况、流式细胞术检测凋亡率、Western blot检测HMGB1和自噬有关蛋白的表达水平;双荧光素酶报告实验评价miR-142-3p对HMGB1的靶向调控作用。结果阿霉素耐药株MCF-7/DOX细胞中的miR-142-3p水平明显下调。miR142-3p的过度表达增强了乳腺癌细胞对阿霉素的敏感性和提高阿霉素诱导的凋亡比率。HMGB1是乳腺癌细胞中miR-142-3p的直接功能靶点,HMGB1的过表达可以明显解除由miR-142-3p上调所产生的细胞凋亡和自噬抑制。结论miR-142-3p的过表达可能通过抑制自噬靶向HMGB1,增强乳腺癌细胞对阿霉素的化学敏感性。miR-142-3p/HMGB1为提高乳腺癌对药物的敏感性提供了新的靶点,具有一定价值。
文摘Our previous studies indicate that phosphatidylinositol 4-kinase Ila can promote the growth of multi-malignant tumors via HER-2/PI3K and MAPK pathways. However, the molecular mechanisms of this pathway and its potential for clinical application remain unknown. In this study, we found that PI4KIla could be an ideal combinatorial target for EGFR treatment via regulating EGFR degradation. Results showed that PI4KIla knockdown reduced EGFR protein level, and the expression of Pi4KIla shows a strong correlation with EGFR in human breast cancer tissues （r= 0.77, P 〈 0.01）. PI4KIla knockdown greatly prolonged the effects and decreased the effective dosage of AG-1478, a specific inhibitor of EGFR. In addition, it significantly enhanced AG1478-induced inhibition of tumor cell survival and strengthened the effect of the EGFR-targeting anti-cancer drug Iressa in xenograft tumor models. Mechanistically, we found that PI4KIla suppression increased EGFR ligand-independent degradation. Quantitative proteomic analysis by stable isotope labeling with amino acids in cell culture （SlLAC） and LC-MS/MS suggested that HsPg0 mediated the effect of PI4KIla on EGFR. Furthermore, we found that combined inhibition of PI4KIla and EGFR suppressed both PI3K/AKT and MAPK/ERK pathways, and resulted in downregulation of multiple oncogenes like PRDX2, FASN, MTA2, ultimately leading to suppression of tumor growth. Therefore, we conclude that combined inhibition of PI4KIla and EGFR exerts a multiple anti-tumor effect. Dual inhibition of EGFR at protein and activity level via combinatorial blocking of PI4KIla presents a novel strategy to combat EGFR-dependent tumors.
