目的:观察乳腺珠蛋白(mammaglobin A,MGBA)负载脐带血来源树突状细胞(dendritic cell,DC)诱导产生的细胞毒性T细胞(cytotoxic T lymphocyte,CTL)对乳腺癌细胞的体外杀伤效果。方法:采集健康剖宫产女性志愿者的脐带血,分离脐带血单个核...目的:观察乳腺珠蛋白(mammaglobin A,MGBA)负载脐带血来源树突状细胞(dendritic cell,DC)诱导产生的细胞毒性T细胞(cytotoxic T lymphocyte,CTL)对乳腺癌细胞的体外杀伤效果。方法:采集健康剖宫产女性志愿者的脐带血,分离脐带血单个核细胞并诱导DC生成,用MGBA致敏DC与自体淋巴细胞共培养诱导CTL。采用流式细胞术测定DC的表型(CD83、CD86、HLADR)变化,ELISA法测定IL-10、IL-12分泌水平,CCK-8法测定CTL对乳腺癌细胞的杀伤活性。结果:成功培养出形态典型、功能成熟的DC,MGBA负载DC诱导的MGBA特异性CTL对乳腺癌细胞MDA-MB-415产生显著的杀伤效果(P<0.05);加入HLA-I抗体可显著减弱杀伤效果,加入HLA-II抗体细胞毒活性无显著变化,加入HLA-I抗体或HLA-II抗体对正常乳腺细胞的杀伤性均无影响。结论:MGBA能明显加强脐带血DC诱导的CTL对乳腺癌细胞的杀伤活性,该杀伤活性具有MHC限制性。展开更多
目的研究人参皂甙Rg3是否是通过PI3K/Akt信号通路来抑制MGBA的表达,进而促进乳腺癌MDA-MB-231细胞的凋亡。方法实验分组:40 nmol/L IGF-1组、100 ng/m L Rg3组、IGF-1+Rg3组和空白对照组。Western-blot检测人参皂苷Rg3对乳腺癌MDA-MB-23...目的研究人参皂甙Rg3是否是通过PI3K/Akt信号通路来抑制MGBA的表达,进而促进乳腺癌MDA-MB-231细胞的凋亡。方法实验分组:40 nmol/L IGF-1组、100 ng/m L Rg3组、IGF-1+Rg3组和空白对照组。Western-blot检测人参皂苷Rg3对乳腺癌MDA-MB-231细胞内p-Akt及MGBA蛋白表达的影响。MTT法检测作用48 h后人参皂苷Rg3对乳腺癌MDA-MB-231细胞增殖的抑制程度;流式细胞术检测作用48 h后人参皂苷Rg3对乳腺癌MDA-MB-231细胞凋亡的影响。结果 Trans well检测结果显示:Rg3可以抑制乳腺癌MDA-MB-231细胞中MGBA的表达,但当增强p-Akt的表达时Rg3的这种抑制作用被明显降低;MTT法检测结果显示:与空白对照组比较,IGF-1组和IGF-1+Rg3组明显促进细胞增殖,而Rg3组明显抑制细胞增殖;流式细胞术检测结果显示:与空白对照组比较,IGF-1组和IGF-1+Rg3组细胞凋亡率明显降低,而Rg3组细胞凋亡率明显增强。结论人参皂甙Rg3干扰乳腺癌细胞内MGBA的表达来促进乳腺癌细胞凋亡的作用是通过影响PI3K/Akt信号通路活性来得以实现的。展开更多
Background and Objectives: Breast cancer is among the most common causes of cancer related mortality in women worldwide. Early detection and prompt diagnosis of tumor is the first step to prevent cancer-related morbid...Background and Objectives: Breast cancer is among the most common causes of cancer related mortality in women worldwide. Early detection and prompt diagnosis of tumor is the first step to prevent cancer-related morbidity and mortality, and a comprehensive understanding of the involved molecular mechanisms can greatly help in this respect. Breast cancer, like many other types of cancer, is caused by a combination of genetic and epigenetic changes such as inactivation of tumor suppressor genes. Materials and Methods: This study was performed on 40 breast cancer patients and 40 healthy controls. Quantitative real time reverse transcription polymerase chain reaction (real time qRT-PCR) was used to assess the expression of carcinoembryonic antigen (CEA) and mammaglobin mRNA in the peripheral blood of patients and healthy controls. The two groups were compared using t-test. Results: The two groups were not significantly different in terms of the mean age. Twenty-nine out of 40 cancer patients were positive for CEA mRNA and its sensitivity was calculated to be 72.5%. Twelve out of 40 healthy controls were positive for CEA mRNA. Twenty-six out of 40 patients were positive for mammaglobin mRNA indicative of 65% sensitivity while only five out of 40 healthy controls were positive for mammaglobin mRNA. Conclusion: Both CEA and mammaglobin mRNA had high sensitivity in cancer patients;thus, they can be used for screening and early detection of breast cancer patients. Further studies with larger sample sizes are required to confirm the current findings.展开更多
Objective: The aim of the study was to explore the individual detection significances of small breast epithelial mucin (SBEM) and human mammaglobin (hMAM) in peripheral blood (PB) of breast cancer patients. Met...Objective: The aim of the study was to explore the individual detection significances of small breast epithelial mucin (SBEM) and human mammaglobin (hMAM) in peripheral blood (PB) of breast cancer patients. Methods: SBEM and hMAM expressions in PB samples of 109 primary breast cancer patients were detected by flow cytometry (FCM) and RT- PCR. Relationship between the biomarkers' expression and prognostic parameters were analyzed. Results: SBEM and hMAM expressions in PB of breast cancer patients were much higher than those of healthy donors and other cancer patients. SBEM and hMAM expressed in 53.2% (50/94) and 39.4% (37/94) cases at stages I-III and expressed in 73.3% (11/15) and 46.7% (7/15) cases at stage IV respectively. SBEM and hMAM mRNA were only detected in PB samples of breast cancer patients, while no expression of them was found in that of healthy donors and other cancer patients. Conclusion: hMAM mRNA detection maybe helpful to predict hematogenous micrometastasis in ER-positive, well-differentiated breast cancers and SBEM mRNA detection maybe helpful to predict hematogenous micrometastasis in ER-negative, poody-differentiated breast cancers.展开更多
目的克隆人乳腺珠蛋白(hum an m amm aglob in,hM aM)基因,建立荧光定量PCR方法,探讨外周血hM aM mRNA的表达作为乳腺癌血行微小转移标志物的可能性。方法设计特异引物,用RT-PCR方法从乳腺癌组织扩增获得目的片段,利用T/A克隆将PCR产物...目的克隆人乳腺珠蛋白(hum an m amm aglob in,hM aM)基因,建立荧光定量PCR方法,探讨外周血hM aM mRNA的表达作为乳腺癌血行微小转移标志物的可能性。方法设计特异引物,用RT-PCR方法从乳腺癌组织扩增获得目的片段,利用T/A克隆将PCR产物插入pMD-18T载体;优化扩增条件,以pMD/M质粒制备标准曲线,建立FQ-PCR方法,对乳腺癌不同分期的患者(n=28),乳腺良性疾病患者(n=15),其他肿瘤患者(n=33)的外周血进行检测。结果成功获得hM aM基因并完成克隆,阳性克隆测序结果与预期序列一致;建立了FQ-PCR检测hM aM mRNA方法;28例乳腺癌病人中9例(32.1%)阳性(Ⅰ期4例,Ⅱ期3例,Ⅲ期1例,Ⅳ期1例),hM aM mRNA的表达与乳腺癌病理分期无关(χ2=0.5936;P>0.05);乳腺良性疾病组,其他肿瘤病人组及对照组外周血中均未检出hM aM mRNA。结论hM aM mRNA是乳腺癌特异性标志物,其外周血的表达可作为乳腺癌血行微小转移的指标。展开更多
文摘目的:观察乳腺珠蛋白(mammaglobin A,MGBA)负载脐带血来源树突状细胞(dendritic cell,DC)诱导产生的细胞毒性T细胞(cytotoxic T lymphocyte,CTL)对乳腺癌细胞的体外杀伤效果。方法:采集健康剖宫产女性志愿者的脐带血,分离脐带血单个核细胞并诱导DC生成,用MGBA致敏DC与自体淋巴细胞共培养诱导CTL。采用流式细胞术测定DC的表型(CD83、CD86、HLADR)变化,ELISA法测定IL-10、IL-12分泌水平,CCK-8法测定CTL对乳腺癌细胞的杀伤活性。结果:成功培养出形态典型、功能成熟的DC,MGBA负载DC诱导的MGBA特异性CTL对乳腺癌细胞MDA-MB-415产生显著的杀伤效果(P<0.05);加入HLA-I抗体可显著减弱杀伤效果,加入HLA-II抗体细胞毒活性无显著变化,加入HLA-I抗体或HLA-II抗体对正常乳腺细胞的杀伤性均无影响。结论:MGBA能明显加强脐带血DC诱导的CTL对乳腺癌细胞的杀伤活性,该杀伤活性具有MHC限制性。
文摘目的研究人参皂甙Rg3是否是通过PI3K/Akt信号通路来抑制MGBA的表达,进而促进乳腺癌MDA-MB-231细胞的凋亡。方法实验分组:40 nmol/L IGF-1组、100 ng/m L Rg3组、IGF-1+Rg3组和空白对照组。Western-blot检测人参皂苷Rg3对乳腺癌MDA-MB-231细胞内p-Akt及MGBA蛋白表达的影响。MTT法检测作用48 h后人参皂苷Rg3对乳腺癌MDA-MB-231细胞增殖的抑制程度;流式细胞术检测作用48 h后人参皂苷Rg3对乳腺癌MDA-MB-231细胞凋亡的影响。结果 Trans well检测结果显示:Rg3可以抑制乳腺癌MDA-MB-231细胞中MGBA的表达,但当增强p-Akt的表达时Rg3的这种抑制作用被明显降低;MTT法检测结果显示:与空白对照组比较,IGF-1组和IGF-1+Rg3组明显促进细胞增殖,而Rg3组明显抑制细胞增殖;流式细胞术检测结果显示:与空白对照组比较,IGF-1组和IGF-1+Rg3组细胞凋亡率明显降低,而Rg3组细胞凋亡率明显增强。结论人参皂甙Rg3干扰乳腺癌细胞内MGBA的表达来促进乳腺癌细胞凋亡的作用是通过影响PI3K/Akt信号通路活性来得以实现的。
文摘Background and Objectives: Breast cancer is among the most common causes of cancer related mortality in women worldwide. Early detection and prompt diagnosis of tumor is the first step to prevent cancer-related morbidity and mortality, and a comprehensive understanding of the involved molecular mechanisms can greatly help in this respect. Breast cancer, like many other types of cancer, is caused by a combination of genetic and epigenetic changes such as inactivation of tumor suppressor genes. Materials and Methods: This study was performed on 40 breast cancer patients and 40 healthy controls. Quantitative real time reverse transcription polymerase chain reaction (real time qRT-PCR) was used to assess the expression of carcinoembryonic antigen (CEA) and mammaglobin mRNA in the peripheral blood of patients and healthy controls. The two groups were compared using t-test. Results: The two groups were not significantly different in terms of the mean age. Twenty-nine out of 40 cancer patients were positive for CEA mRNA and its sensitivity was calculated to be 72.5%. Twelve out of 40 healthy controls were positive for CEA mRNA. Twenty-six out of 40 patients were positive for mammaglobin mRNA indicative of 65% sensitivity while only five out of 40 healthy controls were positive for mammaglobin mRNA. Conclusion: Both CEA and mammaglobin mRNA had high sensitivity in cancer patients;thus, they can be used for screening and early detection of breast cancer patients. Further studies with larger sample sizes are required to confirm the current findings.
文摘Objective: The aim of the study was to explore the individual detection significances of small breast epithelial mucin (SBEM) and human mammaglobin (hMAM) in peripheral blood (PB) of breast cancer patients. Methods: SBEM and hMAM expressions in PB samples of 109 primary breast cancer patients were detected by flow cytometry (FCM) and RT- PCR. Relationship between the biomarkers' expression and prognostic parameters were analyzed. Results: SBEM and hMAM expressions in PB of breast cancer patients were much higher than those of healthy donors and other cancer patients. SBEM and hMAM expressed in 53.2% (50/94) and 39.4% (37/94) cases at stages I-III and expressed in 73.3% (11/15) and 46.7% (7/15) cases at stage IV respectively. SBEM and hMAM mRNA were only detected in PB samples of breast cancer patients, while no expression of them was found in that of healthy donors and other cancer patients. Conclusion: hMAM mRNA detection maybe helpful to predict hematogenous micrometastasis in ER-positive, well-differentiated breast cancers and SBEM mRNA detection maybe helpful to predict hematogenous micrometastasis in ER-negative, poody-differentiated breast cancers.
文摘目的克隆人乳腺珠蛋白(hum an m amm aglob in,hM aM)基因,建立荧光定量PCR方法,探讨外周血hM aM mRNA的表达作为乳腺癌血行微小转移标志物的可能性。方法设计特异引物,用RT-PCR方法从乳腺癌组织扩增获得目的片段,利用T/A克隆将PCR产物插入pMD-18T载体;优化扩增条件,以pMD/M质粒制备标准曲线,建立FQ-PCR方法,对乳腺癌不同分期的患者(n=28),乳腺良性疾病患者(n=15),其他肿瘤患者(n=33)的外周血进行检测。结果成功获得hM aM基因并完成克隆,阳性克隆测序结果与预期序列一致;建立了FQ-PCR检测hM aM mRNA方法;28例乳腺癌病人中9例(32.1%)阳性(Ⅰ期4例,Ⅱ期3例,Ⅲ期1例,Ⅳ期1例),hM aM mRNA的表达与乳腺癌病理分期无关(χ2=0.5936;P>0.05);乳腺良性疾病组,其他肿瘤病人组及对照组外周血中均未检出hM aM mRNA。结论hM aM mRNA是乳腺癌特异性标志物,其外周血的表达可作为乳腺癌血行微小转移的指标。