Application of microarrays in screening the tumor-specific genes in the genome of K562 cells

Abstract

Abstract: Objective To screen tumor-specific genes of K562 cells using DNA microarray technique. Methods The genomic DNA of normal white blood cells and cultured K562 cells were respectively purified and digested with Sau3AⅠ, and the digested DNA fragments of K562 cells were cloned into TA cloning vector to construct the corresponding genomic DNA library. The insert genomic DNA fragments were amplified from the library to prepare the microarray using Cartesian 5500 Microarrayer. The digested genomic DNA fragments of normal white blood cells were labeled with fluorescent Cy3 by restriction display PCR (RD-PCR), followed by hybridization with the microarray, after which the slide was washed and scanned with ScanArray. Results Among the 426 target genes, 42 differential genes were identified in the genomic DNA of K562 cells in comparison with the normal white blood cells. One of the genes was identified as the breakpoint cluster gene (BCR) after sequence analysis. Conclusions The DNA microarrays we constructed may effectively identify the tumor-specific genes in K562 cells, and DNA microarray technique can be helpful in elucidating the molecular mechanisms of tumorigenesis at the genomic level.

CLC Number:

R378.61

PENG Gui-fu¹, MA Wen-li¹, SONG Yan-bing¹, PENG Yi-fei¹, SHI Rong¹, MAO Xiang-ming¹, ZHENG Wen-ling². Application of microarrays in screening the tumor-specific genes in the genome of K562 cells[J]. Journal of Southern Medical University, 2004, 24(05): 525-528.

References

[1] 马文丽,郑文岭,JamesFB,等.限制性显示(RD-PCR):一种新的差异显示技术[A].见:孙志贤.全军生物化学与分子生物学研究进展[M].北京:军事医学科学出版社,1998.89-113
[2] 赵伟,刘全俊,刘伟,等.DNA芯片技术诊断乙、丙型病毒性肝炎的研究[J].东南大学学报(医学版),2002,21(1):75-80.Zhao W, Liu QJ, Liu W, et al. Clinical application of DNA microarray technique to recognition of hepatitis B and C [J]. J Southeast Univ (Medical Science Edition), 2002, 21(1): 75-80.
[3] 姜立,马文丽人红白血病K562细胞与红系分化的调控[J].第一军医大学学报(J First Mil Med Univ/Di Yi Jun Yi Da Xue Xue Bao),2001,21(6):461-3.
[4] 陈汉春,孟巧.K562细胞中氯化血红素诱导性表达基因的研究[J].中华血液学杂志,2003,24(4):185-9.Chen HC, Meng Q. Study ofhemin induced gene express in k562 cells[J]. Chin J Hematol, 2003, 24 (4): 185-9.
[5] 冯春琼,马文丽,李凌,等.As₂O₃ 处理前后K562细胞基因表达变化的基因芯片检测[J].第一军医大学学报,2002,22(9):772-5.Feng CQ, Ma WL, Li L, et al. Analysis with DNA chips of the gene expression in K562 cells in response to As₂O₃ treatment[J]. J First Mil Mea Univ/Di Yi Jun Yi Da Xue Xue Bao, 2002, 22(9): 772-5.
[6] 朱利娜,马文丽,毛向明,等.人胎盘基因表达谱芯片的初步研究[J].第一军医大学学报,2002,22(5):400-2.Zhu LN, Ma WL, Mao XM, et al. Preliminary study of DNA microarray of human placenta gene expression profile [J]. J First Mil Med Univ/Di Yi Jun Yi Da Xue Xue Bao, 2002, 22 (5): 400-2.
[7] 毛向明,马文丽,姜立,等.应用RD-PCR方法制备K562细胞表达谱芯片基因探针[J].第一军医大学学报,2002,22(6):548-53.Mao XM, Ma WL, Jiang L, et al. Application of restriction display PCR in preparing the probes of the gene chip for investigating gene expression profile of K562 cells [J]. J First Mil Med Univ/Di Yi Jun Yi Da Xue Xue Bao, 2002, 22(6): 548-53.
[8] Lajos P, Mark A, James S, et al. Clinical application of cDNA microarrays in oncology[J]. Oncology, 2003, 8(2): 252-8.
[9] 张强,朱志文.抑制性消减杂交技术与肿瘤相关基因克隆新进展[J].国外医学·肿瘤学分册(Foreign Med Sci·Section of Onco1ogy),2000,27(6):362-4.
[10] 孙立军,黄强,王爱东,等.应用差异显示技术克隆胶质瘤细胞诱导分化相关基因[J].中华神经外科杂志,2003,19(2):89-92.Sun LJ, Huang Q, Wang AD, et al. Clone the genes associated with induced differentiation of glioma with RAP-DD [J]. Chin J Neurosurg, 2003, 19(2): 89-92.
[12] Daley GQ, van Etten RA, Baltimore D. Induction of chronic myelogenous leukemia in mice by the P210 bcr/abl gene of the Philadelphia chromosome [J]. Science, 1990, 247(243): 824-30.
[13] 王春红,孙秉中,袁跃传.bcr-abl融合基因抑制K562细胞凋亡的研究[J].中华肿瘤杂志,1998,20(5):340-1.Wang CH, Sun BZ, Yuan YC. Inhibition of apoptosis by bcr-abl fusion gene in k562 cells[J]. Chin J Oncol, 1998, 20(5): 340-1.
[11] Ciro M, Bracken AP, Helin K. Profiling cancer [J]. Curr Opin Cell Biol, 2003, 15(2): 213-20.