[应用抑制性消减杂交技术鉴定白血病多药耐药相关基因]
[Identification of multidrug resistance related genes in leukemia by suppression subtractive hybridization].
作者信息
Zhu Ning-xi, Zheng Shu, Xu Rong-zhen, Gao Rui-lan, Shen Jian-ping, Yu Rong-xi
机构信息
Zhejiang Provincial Hospital of Traditional Chinese Medicine, Hangzhou 310006, China.
出版信息
Zhonghua Xue Ye Xue Za Zhi. 2003 Jan;24(1):14-7.
OBJECTIVE
To clone and screen genes related to multidrug resistance (MDR) in leukemia.
METHODS
Suppression subtractive hybridization (SSH) was performed to profile differentially expressed genes between a MDR leukemia cell line (K562/DOX, as tester) and its parent cell line (K562, as driver). Reverse Northern dot blot was carried out to further screen the subtracted cDNA library. The overexpressed cDNA fragments in K562/DOX cells were sequenced and compared with known genes in Genbank. RT-PCR and Northern blot were employed to confirm the differential expression of some identified genes.
RESULTS
Eleven genes were identified being overexpressed in K562/DOX, including S3 ribosomal protein (S3rp) gene, NADH dehydrogenase subunit 2 (ND2) gene and My023 gene, which have not been reported to be related to MDR in cancer.
CONCLUSION
Several genes, which might be involved in MDR were identified, indicating novel mechanisms of MDR in leukemia.
目的
克隆并筛选白血病中与多药耐药（MDR）相关的基因。
方法
采用抑制性消减杂交（SSH）技术分析多药耐药白血病细胞系（K562/DOX，作为检测子）与其亲本细胞系（K562，作为驱动子）之间差异表达的基因。进行反向Northern斑点杂交进一步筛选消减cDNA文库。对K562/DOX细胞中高表达的cDNA片段进行测序，并与Genbank中的已知基因进行比较。采用RT-PCR和Northern印迹法确认部分已鉴定基因的差异表达。
结果
鉴定出11个在K562/DOX中高表达的基因，包括S3核糖体蛋白（S3rp）基因、NADH脱氢酶亚基2（ND2）基因和My023基因，这些基因尚未见在癌症中与多药耐药相关的报道。
结论
鉴定出几个可能参与多药耐药的基因，提示白血病多药耐药的新机制。
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