Liu Shi-Qi, Cheng Jing-Ying, Jiang Ya-Jing, Li Ying, Li Qing-Hua, Pang Tian-Xiang
State Key Laboratory of Experimental Hematology, Institute of Hematology and Blood Diseases Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Tianjin 300020, China.
State Key Laboratory of Experimental Hematology, Institute of Hematology and Blood Diseases Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Tianjin 300020, China,E-mail:
Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2019 Apr;27(2):311-317. doi: 10.19746/j.cnki.issn.1009-2137.2019.02.001.
To screen and verify the differentially expressed genes related with aging of bone marrow mesenchymal stem cells (BM-MSCs) in acute myeloid leukemia (AML) patients by bioinformatics, so as to provide new molecular markers for the research and clinical treatment of AML.
The gene expression profiling chip related with BM-MSCs in AML patients in our hospital and the gene chip GSE84881 selected from NCBI database GEO were used for data analysis and exploration. The DAVID analysis software was used to perform gene ontology (GO) enrichment analysis and KEGG pathway enrichment analysis. Furthermore, the differentially expressed genes related with aging of BM-MSCs in AML patients were identified. Bone marrow samples were collected and MSCs were amplified in vitro, and RT-PCR was used to verify the differentially expressed genes, which should be further identified with senescence-associated β-galactosidase staining and MTT cell proliferation assays.
A total of 247 differentially expressed genes were screened out by bioinformatics methods, including genes of 132 up-regulated expression and 115 down-regulated expression. Six differentially expressed genes related with aging of BM-MSCs in AML patients were screened out, including the genes of up-regulated expression, COL3A1 (P<0.05), CRYAB (P<0.01), DCN (P<0.05), and the genes of down-regulated expression, including CCL2 (P<0.05), CTSC (P<0.01) and IL6 (P<0.05). These 6 differentially expressed genes were consistent with data from chip assays, and which was significantly correlated with aging of BM-MSCs in AML patients. Meanwhile, the positive rate of senescence-associated β-galactosidase staining in BM-MSCs of AML patients was significantly different from that of healthy donors (P<0.01). MTT cell proliferation assay showed that BM-MSCs in AML patients had proliferative ability lower than the healthy donors' BM-MSCs.
The data here suggest novel clues for the clinical research and treatment of BM-MSCs aging in AML patients.
通过生物信息学方法筛选并验证急性髓系白血病（AML）患者中与骨髓间充质干细胞（BM-MSCs）衰老相关的差异表达基因，为AML的研究及临床治疗提供新的分子标志物。
采用我院AML患者中与BM-MSCs相关的基因表达谱芯片以及从NCBI数据库GEO中选取的基因芯片GSE84881进行数据分析与探索。运用DAVID分析软件进行基因本体（GO）富集分析和KEGG通路富集分析。此外，鉴定出AML患者中与BM-MSCs衰老相关的差异表达基因。采集骨髓样本并体外扩增MSCs，采用RT-PCR验证差异表达基因，并用衰老相关β-半乳糖苷酶染色和MTT细胞增殖试验进一步鉴定。
通过生物信息学方法共筛选出247个差异表达基因，其中上调表达基因132个，下调表达基因115个。筛选出6个与AML患者BM-MSCs衰老相关的差异表达基因，上调表达的基因有COL3A1（P<0.05）、CRYAB（P<0.01）、DCN（P<0.05），下调表达的基因有CCL2（P<0.05）、CTSC（P<0.01）和IL6（P<0.05）。这6个差异表达基因与芯片检测数据一致，且与AML患者BM-MSCs衰老显著相关。同时，AML患者BM-MSCs中衰老相关β-半乳糖苷酶染色阳性率与健康供体有显著差异（P<0.01）。MTT细胞增殖试验显示，AML患者的BM-MSCs增殖能力低于健康供体的BM-MSCs。
本研究数据为AML患者BM-MSCs衰老的临床研究和治疗提供了新线索。
