Growth arrest specific 2 is up-regulated in chronic myeloid leukemia cells and required for their growth.
Although the generation of BCR-ABL is the molecular hallmark of chronic myeloid leukemia (CML), the comprehensive molecular mechanisms of the disease remain unclear yet. Growth arrest specific 2 (GAS2) regulates multiple cellular functions including cell cycle, apoptosis and calpain activities. In the present study, we found GAS2 was up-regulated in CML cells including CD34+ progenitor cells compared to their normal counterparts. We utilized RNAi and the expression of dominant negative form of GAS2 (GAS2DN) to target GAS2, which resulted in calpain activity enhancement and growth inhibition of both K562 and MEG-01 cells. Targeting GAS2 also sensitized K562 cells to Imatinib mesylate (IM). GAS2DN suppressed the tumorigenic ability of MEG-01 cells and impaired the tumour growth as well. Moreover, the CD34+ cells from CML patients and healthy donors were transduced with control and GAS2DN lentiviral vectors, and the CD34+ transduced (YFP+) progeny cells (CD34+YFP+) were plated for colony-forming cell (CFC) assay. The results showed that GAS2DN inhibited the CFC production of CML cells by 57±3% (n = 3), while affected those of normal hematopoietic cells by 31±1% (n = 2). Next, we found the inhibition of CML cells by GAS2DN was dependent on calpain activity but not the degradation of beta-catenin. Lastly, we generated microarray data to identify the differentially expressed genes upon GAS2DN and validated that the expression of HNRPDL, PTK7 and UCHL5 was suppressed by GAS2DN. These 3 genes were up-regulated in CML cells compared to normal control cells and the growth of K562 cells was inhibited upon HNRPDL silence. Taken together, we have demonstrated that GAS2 is up-regulated in CML cells and the inhibition of GAS2 impairs the growth of CML cells, which indicates GAS2 is a novel regulator of CML cells and a potential therapeutic target of this disease.
The First Affiliated Hospital, Jiangsu Institute of Hematology, Key Laboratory of Thrombosis and Hemostasis of Ministry of Health, Soochow University, Suzhou, Jiangsu Province, P.R. China ; Cyrus Tang Hematology Center, Soochow University, Suzhou, Jiangsu Province, P.R. China.
Haixia
H
Zhou
Cyrus Tang Hematology Center, Soochow University, Suzhou, Jiangsu Province, P.R. China.
Yue
Y
Ge
Cyrus Tang Hematology Center, Soochow University, Suzhou, Jiangsu Province, P.R. China.
Lili
L
Sun
Cyrus Tang Hematology Center, Soochow University, Suzhou, Jiangsu Province, P.R. China.
Wenjuan
W
Ma
Cyrus Tang Hematology Center, Soochow University, Suzhou, Jiangsu Province, P.R. China.
Jie
J
Wu
Cyrus Tang Hematology Center, Soochow University, Suzhou, Jiangsu Province, P.R. China.
Xiuyan
X
Zhang
The First Affiliated Hospital, Jiangsu Institute of Hematology, Key Laboratory of Thrombosis and Hemostasis of Ministry of Health, Soochow University, Suzhou, Jiangsu Province, P.R. China.
Xiaohui
X
Hu
Terry Fox Laboratory, British Columbia Cancer Agency, Vancouver, Canada.
Connie J
CJ
Eaves
The First Affiliated Hospital, Jiangsu Institute of Hematology, Key Laboratory of Thrombosis and Hemostasis of Ministry of Health, Soochow University, Suzhou, Jiangsu Province, P.R. China.
Depei
D
Wu
Cyrus Tang Hematology Center, Soochow University, Suzhou, Jiangsu Province, P.R. China.
Yun
Y
Zhao
D000818:Animals; D002154:Calpain / Q000378:metabolism; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D020869:Gene Expression Profiling; D015973:Gene Expression Regulation, Leukemic; D055785:Gene Knockdown Techniques; D006801:Humans; D015464:Leukemia, Myelogenous, Chronic, BCR-ABL Positive / Q000235:genetics* / Q000473:pathology*; D051379:Mice; D008819:Mice, Nude; D008840:Microfilament Proteins / Q000378:metabolism*; D014411:Neoplastic Stem Cells / Q000378:metabolism / Q000473:pathology; D034622:RNA Interference; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D059467:Transcriptome / Q000235:genetics; D014161:Transduction, Genetic; D014410:Tumor Stem Cell Assay; D015854:Up-Regulation; D051176:beta Catenin / Q000378:metabolism
C490936:GAS2 protein, human; D008840:Microfilament Proteins; D012333:RNA, Messenger; D051176:beta Catenin; D002154:Calpain; C569870:CAPN2 protein, human