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基因芯片技术筛查并鉴定乳腺癌细胞中MAGE-A11的相关基因[J].谷丽娜,桑梅香,李娟,刘飞,王芃堉,尹丹静,吴云艳,单保恩.中国肿瘤生物治疗杂志,2018,(9):904~912.
基因芯片技术筛查并鉴定乳腺癌细胞中MAGE-A11的相关基因    点此下载全文    点此浏览HTML全文
谷丽娜  桑梅香  李娟  刘飞  王芃堉  尹丹静  吴云艳  单保恩
河北医科大学 第四医院a. 科研中心;b. 肿瘤研究所免疫室,河北石家庄050011
基金项目:河北省科技计划资助项目(No.152777184);河北省杰出青年基金资助项目(No.H2016206410);河北省财政厅资助项目(No.[2016]361006)
DOI:10.3872/j.issn.1007-385X.2018.09.010
摘要:
      目的:应用高通量基因芯片技术筛查乳腺癌细胞中黑色素瘤相关抗原(melanoma antigen,MAGE)-A11 的相关基因,并从数量和功能两方面加以验证。方法:采用基因芯片技术筛选乳腺癌MCF-7、MDA-MB-231 和BT-549 中MAGE-A11 下游靶基因的mRNA的差异表达,对有代表性的基因进行了聚类分析,并利用qRT-PCR进行验证。以CCK-8 法、细胞划痕实验和Transwell实验检测MAGE-A11 对乳腺癌细胞中增殖、迁移和侵袭功能的影响。结果:3 种乳腺癌细胞过表达MAGE-A11 导致1 608个下游基因差异表达,主要涉及蛋白泛素化、细胞增殖和凋亡、肿瘤侵袭和转移。基因芯片中典型高表达的ZNF-451、CENPTJ、CDK13、API5 和LMO7 在qRT-PCR 在验证结果中也显著高于对照组(P<0.01),低表达的SHPRH、PML、MARK2、LIMA1 和ANGPTL4也显著低于对照组(P<0.01)。转染MAGE-A11 组的乳腺癌细胞MCF-7、MDA-MB-231 和BT-549 72 h 的增殖能力较对照组明显增强(均P<0.01),培养48 h 后与对照组相比,转染MAGE-A11 的3 种细胞划痕出现明显愈合(P<0.05 或P<0.01),穿膜数较对照组明显增多(均P<0.01)。结论:在MCF-7、MDA-MB-231 和BT-549 三种乳腺癌细胞中筛查到涉及蛋白泛素化、细胞增殖和凋亡、肿瘤侵袭和迁移等生物功能众多的表达差异基因,对其中10 种典型差异基因从数量和功能两方面进行验证,并得到初步确认。
关键词:乳腺癌  黑色素瘤相关抗原-A11  基因芯片
Screening and identification of MAGE-A11 related genes based on DNAmicroarray    Download Fulltext
GU Lina  SANG Meixiang  LI Juan  LIU Fei  WANG Pengyu  YIN Danjing  WU Yunyan  SHAN Baoen
a.Department of Research Center;b. Department of Immunology, Tumor Research Institute, the Fourth Hospital of Hebei Medical University, Shijiazhuang 050011, Hebei, China
Fund Project:Project supported by the Science and Technology Program of Hebei Province ( No.152777184), the Foundation for Distinguished Young-Scholars of Hebei Provience(No. H2016206410), and the Supporting Program from Financial Department of Hebei Province(No. [2016] 361006)
Abstract:
      Objective: To screen related genes of melanoma-associated antigen-A11 (MAGE-A11) in breast cancer cells based on highthroughput DNA microarray technology, and to validate from the aspects of quantity and function. Methods: DNA microarray was used to screen the differently-expresseddown-stream mRNAs of MAGE-A11 in breast cancercelllines (MCF-7, MDA-MB-231 and BT-549).Cluster analysis was applied on representative genes and quantitative RT-PCR was used to validate. CCK-8, scratch wound healing assay and Transwell assaywere used to detect the effect of MAGE-A11 on the proliferation,migration and invasion of breast cancer cells.Results: Over-expression of MAGE-A11 caused the differential expression of 1608 down-stream genes in 3 breast cancer cell lines,which was associated with various cell functions such as protein ubiquitination,cell proliferation and apoptosis, tumor invasion and metastasis.qRT-PCR validated that the expression of ZNF-451, CENPTJ, CDK13, API5 and LMO7, which were highly expressed in microarray,were also significantly higher than those in control group (P<0.01);in addition, SHPRH, PML, MARK2, LIMA1 and ANGPTL4,which were low-expressed in microarray, were also significantly lower than those in control group (P<0.01). MAGE-A11transfection directly increased the proliferation of breast cancer MCF-7, MDA-MB-231 and BT-549 cells at 72 h (all P<0.01); compared with control group after transfectionexhibited obvious wound healing at 48 h (P<0.05 or P<0.01) and significantly increased trans-membrane cell numbers (all P<0.01). Conclusion: Many differentially expressed genes related to ubiquitination, cell proliferation and apoptosis, tumor invasion and migration were screened in MCF-7, MDA-MB-231 and BT-549 breast cancer cells. Among them, 10 typical differentially expressed genes were identified in terms of quantity and function.
Keywords:breast cancer  melanoma antigen-A11(MAGE-A11)  DNA microarray
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