CART1在乳腺癌中的表达及其初步分子机制研究
发布时间：2019-06-05 12:18
【摘要】：目的:检测软骨同源蛋白1(cartilage hemeprotein 1,CART1)在乳腺癌及癌旁组织中的表达水平,探讨CART1的表达与乳腺癌病理特征的关系。运用外源性RNA干扰技术,敲低MDA-MB-231乳腺癌细胞中软骨同源蛋白1(CART1)基因的表达,研究CART1基因沉默后对MDA-MB-231细胞生物学行为的影响。方法:应用免疫组织化学法检测CART1在乳腺癌组织中的表达,并统计分析CART1的表达量与乳腺癌患者的年龄肿瘤大小、病理分型、组织类型以及临床分期的关系。采用人工合成的针对CART1基因的si RNA,运用转染试剂RNAi MAX转染MDA-MB-231细胞。实时定量PCR检测转染后CART1 m RNA水平,Western blot法检测转染后CART1蛋白水平,Transwell TM侵袭实验检测MDA-MB-231细胞侵袭CCK-8法检测细胞增殖、流式细胞术检测细胞周期的变化。结果:在62例乳腺癌组织标本中,CART1在54例组织标本高表达(87.1%);在8例癌旁组织中CART1不表达或者低表达。CART1-si RNA能有效沉默CART1在MDA-MB-231细胞中的表达,CART1沉默后MDA-MB-231细胞侵袭和增殖能力明显下降,S期细胞的比率增高而G2/M细胞比率减少。结论:CART1在乳腺癌组织中高表达。下调MDA-MB-231细胞CART1的表达,可降低其侵袭和增殖能力、诱导S期阻滞。
[Abstract]:Aim: to detect the expression of cartilage homeoprotein 1 (cartilage hemeprotein 1 and Cart 1 in breast cancer and paracancerous tissues, and to explore the relationship between the expression of CART1 and pathological features of breast cancer. The expression of cartilage homeoprotein 1 (CART1) gene in MDA-MB-231 breast cancer cells was decreased by exogenous RNA interference technique, and the effect of CART1 gene silencing on the biological behavior of MDA-MB-231 cells was studied. Methods: the expression of CART1 in breast cancer tissues was detected by immunohistochemistry, and the expression of CART1 and the age of breast cancer patients were statistically analyzed. Relationship between tumor size, pathological classification, tissue type and clinical stage. MDA-MB-231 cells were transfected with synthetic si RNA, targeting CART1 gene by RNAi MAX. Real-time quantitative PCR was used to detect CART1 m RNA level, Western blot assay to detect CART1 protein level, Transwell TM invasion assay to detect MDA-MB-231 cell invasion CCK-8 assay to detect cell proliferation and flow cytometry to detect cell cycle changes. Results: in 62 cases of breast cancer, CART1 was highly expressed in 54 cases (87.1%). CART1 was not expressed or underexpressed in 8 cases of paracancerous tissues. CART1-si RNA could effectively silence the expression of CART1 in MDA-MB-231 cells, and the invasion and proliferation of MDA-MB-231 cells decreased significantly after CART1 silencing. The rate of S phase cells increased while that of G 2 鈮，
本文编号：2493524
[Abstract]:Aim: to detect the expression of cartilage homeoprotein 1 (cartilage hemeprotein 1 and Cart 1 in breast cancer and paracancerous tissues, and to explore the relationship between the expression of CART1 and pathological features of breast cancer. The expression of cartilage homeoprotein 1 (CART1) gene in MDA-MB-231 breast cancer cells was decreased by exogenous RNA interference technique, and the effect of CART1 gene silencing on the biological behavior of MDA-MB-231 cells was studied. Methods: the expression of CART1 in breast cancer tissues was detected by immunohistochemistry, and the expression of CART1 and the age of breast cancer patients were statistically analyzed. Relationship between tumor size, pathological classification, tissue type and clinical stage. MDA-MB-231 cells were transfected with synthetic si RNA, targeting CART1 gene by RNAi MAX. Real-time quantitative PCR was used to detect CART1 m RNA level, Western blot assay to detect CART1 protein level, Transwell TM invasion assay to detect MDA-MB-231 cell invasion CCK-8 assay to detect cell proliferation and flow cytometry to detect cell cycle changes. Results: in 62 cases of breast cancer, CART1 was highly expressed in 54 cases (87.1%). CART1 was not expressed or underexpressed in 8 cases of paracancerous tissues. CART1-si RNA could effectively silence the expression of CART1 in MDA-MB-231 cells, and the invasion and proliferation of MDA-MB-231 cells decreased significantly after CART1 silencing. The rate of S phase cells increased while that of G 2 鈮，
本文编号：2493524
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