Zhonghua yi xue za zhi · 2004
[Transcriptome profiling of marrow mononuclear cells of patients with myelodysplastic syndrome using cDNA microarray analysis].
Аннотация на английскомСтатьяПоддержка исследования: вне правительства СШАНазвание и аннотация: Перевод в очереди
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Qian Jun, Chen Zi-xing, Wang Wei, Cen Jian-nong, Xue Yong-quan
Аннотация
OBJECTIVE: To study the gene expression profiles of patients with myelodysplastic syndrome (MDS) and try to identify some genes with pathogenetic and diagnostic relevance.
METHODS: The bone marrow mononuclear cells (BMMNCs) of 10 patients with MDS, including 4 cases of refractory anemia (RA), 1 case of refractory thrombocytopenia (RTC), 4 cases of RA with excess blasts (RAEB), and 1 case of RAEB in transformation (RAEBt), were isolated and the total RNA was extracted and underwent transcriptome analysis by using customized cDNA microarray with 500 gene clones. Seventeen specimens of normal bone marrow, as controls, were collected from the ribs of 17 patients with chest tumors. Cluster software was used to make analysis. Semiquantitative RT-PCR was carried out to confirm the results of microarray analysis.
RESULTS: 95 genes, such as the genes of thrombospondin 1 (THBS1), phosphatase and tensin homolog (PTEN), MAX dimerization protein (MAD), DNA-damage-inducible transcript 3 (DDIT3), ets variant gene 1 (ETV1), G1 to S phase transition 1 (GSPT1), were shown to be abnormally expressed in at least 5 MDS patients compared to the normal controls, involving cell growth and differentiation regulation, cell cycle control, signaling, and redox. The 10 MDS patients in different stages were clustered into two distinct groups, whereas a case with refractory thrombocytopenia and other RA patients were clustered into two subgroups. Semiquantitative RT-PCR revealed the identical results in 3 (60%) of the 5 genes determined by microarray analysis. Further analysis on samples from 50 MDS patients confirmed the different expression levels of RNA helicase-related protein (RNAHP), GSPT1, and DDIT3 between the MDS patients and the normal controls.
CONCLUSION: The technology of microarray can reveal the intrinsic molecular features of MDS patients and the detection of DDIT3 and RNAHP expression may be useful for the diagnosis of MDS.
METHODS: The bone marrow mononuclear cells (BMMNCs) of 10 patients with MDS, including 4 cases of refractory anemia (RA), 1 case of refractory thrombocytopenia (RTC), 4 cases of RA with excess blasts (RAEB), and 1 case of RAEB in transformation (RAEBt), were isolated and the total RNA was extracted and underwent transcriptome analysis by using customized cDNA microarray with 500 gene clones. Seventeen specimens of normal bone marrow, as controls, were collected from the ribs of 17 patients with chest tumors. Cluster software was used to make analysis. Semiquantitative RT-PCR was carried out to confirm the results of microarray analysis.
RESULTS: 95 genes, such as the genes of thrombospondin 1 (THBS1), phosphatase and tensin homolog (PTEN), MAX dimerization protein (MAD), DNA-damage-inducible transcript 3 (DDIT3), ets variant gene 1 (ETV1), G1 to S phase transition 1 (GSPT1), were shown to be abnormally expressed in at least 5 MDS patients compared to the normal controls, involving cell growth and differentiation regulation, cell cycle control, signaling, and redox. The 10 MDS patients in different stages were clustered into two distinct groups, whereas a case with refractory thrombocytopenia and other RA patients were clustered into two subgroups. Semiquantitative RT-PCR revealed the identical results in 3 (60%) of the 5 genes determined by microarray analysis. Further analysis on samples from 50 MDS patients confirmed the different expression levels of RNA helicase-related protein (RNAHP), GSPT1, and DDIT3 between the MDS patients and the normal controls.
CONCLUSION: The technology of microarray can reveal the intrinsic molecular features of MDS patients and the detection of DDIT3 and RNAHP expression may be useful for the diagnosis of MDS.