Objective:The aim of this study was to detect the expression level of Metadherin (MTDH) in peripheral blood of the breast cancer patients by real-time fluorescence quantitative polymerase chain reaction (PCR),and to e...Objective:The aim of this study was to detect the expression level of Metadherin (MTDH) in peripheral blood of the breast cancer patients by real-time fluorescence quantitative polymerase chain reaction (PCR),and to explore the relationship between expression of Metadherin gene in the patients peripheral blood and the clinic-pathological features in breast cancer. Methods:Real-time fluorescence quantitative polymerase chain reaction was employed to determine the expression level of Metadherin gene in 80 peripheral blood samples of breast cancer patients and healthy donors. Results:The expression of Metadherin gene in breast cancer patients peripheral blood were positive,in which 34 breast cancer patients were highly expressed,accounting for 55.7%,while the expression of Metadherin gene in normal females peripheral blood were negative,there was statistical significance (Ratio = 2.02±0.81,P < 0.05); Ratio of the Metadherin expression in breast cancer patients peripheral blood and the glyceraldehyde-3-phosphate dehydrogenase expression was 1.15 ± 0.36. REST software analysis showed that the expression of Metadherin gene was significantly up-regulated in breast cancer. Conclusion:The SYBR Green I quantitative real-time polymerase chain reaction method can successfully detect the expression level of Metadherin gene. Expression level of Metadherin gene in breast cancer patients peripheral blood is closely related to survival,and it maybe involved in the development of breast cancer and used as an indicator of prognosis.展开更多
目的建立一种实时荧光定量PCR方法,并用该方法检测乳腺癌患者血浆循环DNA水平。方法用QIAamp血液DNA抽提试剂盒提取61例乳腺癌患者和25例健康妇女血浆循环DNA,用实时荧光定量PCR技术(SYBR Green I染料法)进行定量,结果用设定临界域值时...目的建立一种实时荧光定量PCR方法,并用该方法检测乳腺癌患者血浆循环DNA水平。方法用QIAamp血液DNA抽提试剂盒提取61例乳腺癌患者和25例健康妇女血浆循环DNA,用实时荧光定量PCR技术(SYBR Green I染料法)进行定量,结果用设定临界域值时的循环数的值(Ct值)表示,并对定量结果进行ROC曲线分析。结果:乳腺癌患者Ct值(27.71±1.41)显著低于健康对照(30.37±1.32,P<0.001),说明乳腺癌患者血浆循环DNA水平显著高于健康对照组;ROC曲线下面积为0.921。结论应用实时PCR技术检测乳腺癌患者血浆循环DNA水平有可能成为一种无创性乳腺癌分子诊断方法。展开更多
目的检测RFP(Ret finger protein)蛋白在人正常组织及人癌组织中的分布。方法采用实时荧光定量PCR方法,以18 S rRNA为内对照,检测RFP在人正常组织及人癌组织中的表达水平。结果RFP表达水平子宫颈鳞状细胞癌组织高于正常子宫颈组织,子宫...目的检测RFP(Ret finger protein)蛋白在人正常组织及人癌组织中的分布。方法采用实时荧光定量PCR方法,以18 S rRNA为内对照,检测RFP在人正常组织及人癌组织中的表达水平。结果RFP表达水平子宫颈鳞状细胞癌组织高于正常子宫颈组织,子宫内膜腺癌组织高于正常子宫内膜组织,胃腺癌组织高于正常胃组织,食管鳞状细胞癌组织高于正常食管组织,而子宫内膜腺癌组织高于子宫颈鳞状细胞癌组织,胃腺癌组织高于食管鳞状细胞癌组织,脑癌组织高于正常脑组织。结论RFP可能成为治疗恶性肿瘤的一个潜在的分子靶点。展开更多
文摘Objective:The aim of this study was to detect the expression level of Metadherin (MTDH) in peripheral blood of the breast cancer patients by real-time fluorescence quantitative polymerase chain reaction (PCR),and to explore the relationship between expression of Metadherin gene in the patients peripheral blood and the clinic-pathological features in breast cancer. Methods:Real-time fluorescence quantitative polymerase chain reaction was employed to determine the expression level of Metadherin gene in 80 peripheral blood samples of breast cancer patients and healthy donors. Results:The expression of Metadherin gene in breast cancer patients peripheral blood were positive,in which 34 breast cancer patients were highly expressed,accounting for 55.7%,while the expression of Metadherin gene in normal females peripheral blood were negative,there was statistical significance (Ratio = 2.02±0.81,P < 0.05); Ratio of the Metadherin expression in breast cancer patients peripheral blood and the glyceraldehyde-3-phosphate dehydrogenase expression was 1.15 ± 0.36. REST software analysis showed that the expression of Metadherin gene was significantly up-regulated in breast cancer. Conclusion:The SYBR Green I quantitative real-time polymerase chain reaction method can successfully detect the expression level of Metadherin gene. Expression level of Metadherin gene in breast cancer patients peripheral blood is closely related to survival,and it maybe involved in the development of breast cancer and used as an indicator of prognosis.
文摘本研究拟建立检测CK19基因表达的数字PCR(digital PCR,dPCR)方法并测试其性能,以期利用该方法的超高敏感性和绝对定量优势进行循环肿瘤细胞(circulating tumor cells,CTC)的定量分析。首先,根据CK19基因的mRNA序列进行引物探针设计,以看家基因ABL1作为内参,采用人乳腺癌MCF7细胞和健康人白细胞从13套引物探针之中筛选出了最佳的引物探针,测序结果证实扩增序列无误。其次,针对选定的引物探针进行反应条件的优化,以不同浓度的健康人白细胞cDNA为模板进行空白检测限(limit of blank,LOB)的分析,证实当ABL1基因拷贝数为20,000、15,000、10,000、5000、2500时,CK19的LOB分别为9.24、8.93、3.12、3.17和2.53拷贝。再次,采用MCF7细胞和健康人白细胞的cDNA模板进行不同浓度预混的检测限(limit of detection,LOD)分析,证实50%、10%、5%、1%、0.5%和0.1%的浓度配比下均可以有效检出CK19基因,线性R2值为0.9998。最终,临床样本的数字PCR检测结果发现晚期乳腺癌患者的CK19拷贝数高于健康人对照。上述结果说明本研究所建立的检测CK19基因的数字PCR方法具有敏感、特异和定量准确等优势,未来经过进一步验证之后有望用于乳腺癌的CTC定量分析,具有良好的应用前景。
文摘目的建立一种实时荧光定量PCR方法,并用该方法检测乳腺癌患者血浆循环DNA水平。方法用QIAamp血液DNA抽提试剂盒提取61例乳腺癌患者和25例健康妇女血浆循环DNA,用实时荧光定量PCR技术(SYBR Green I染料法)进行定量,结果用设定临界域值时的循环数的值(Ct值)表示,并对定量结果进行ROC曲线分析。结果:乳腺癌患者Ct值(27.71±1.41)显著低于健康对照(30.37±1.32,P<0.001),说明乳腺癌患者血浆循环DNA水平显著高于健康对照组;ROC曲线下面积为0.921。结论应用实时PCR技术检测乳腺癌患者血浆循环DNA水平有可能成为一种无创性乳腺癌分子诊断方法。
文摘目的检测RFP(Ret finger protein)蛋白在人正常组织及人癌组织中的分布。方法采用实时荧光定量PCR方法,以18 S rRNA为内对照,检测RFP在人正常组织及人癌组织中的表达水平。结果RFP表达水平子宫颈鳞状细胞癌组织高于正常子宫颈组织,子宫内膜腺癌组织高于正常子宫内膜组织,胃腺癌组织高于正常胃组织,食管鳞状细胞癌组织高于正常食管组织,而子宫内膜腺癌组织高于子宫颈鳞状细胞癌组织,胃腺癌组织高于食管鳞状细胞癌组织,脑癌组织高于正常脑组织。结论RFP可能成为治疗恶性肿瘤的一个潜在的分子靶点。
