Knockdown of immature colon carcinoma transcript 1 induces suppression of proliferation, S-phase arrest and apoptosis in leukemia cells

Guang Yao Li, Ji Zhu Liu, Li Zhang, Guo Zhen Liu, Shuang Jing Li, Tai Wu Xiao, Jing Xia Wang, Le Xin Wang, Ming Hou

Research output: Contribution to journalArticle

Abstract

Immature colon carcinoma transcript 1 (ICT1), a human mitochondrial translation release factor, is a ribosome-dependent codon-independent peptidyl-tRNA hydrolase. ICT1-deficiency has been recognized as a cell growth inhibitor of hepatoblastoma and glioblastoma multiforme. To explore the role of ICT1 in human leukemia, 2 short hairpin RNAs (shRNAs) targeting ICT1 sequences were designed in leukemia U937 cells. The successful infection of ICT1 in the U937 cells was observed under a fluorescence microscope and further quantified by western blotting and quantitative real-time PCR (qRT-PCR) analysis. Tetrazolium dye (MTT) assay revealed a significant decrease in proliferation of ICT1-knockdown U937 cells on the fourth and fifth day as compared with the control. Depletion of ICT1 resulted in an increase in S phase and sub-G1 (representing cell apoptosis) fractions. Annexin V-APC/7-AAD staining assay confirmed that knockdown of ICT1 played a crucial role in boosting early and late apoptotic programs in U937 cells. Downregulation of ICT1 also altered cyclin A2 transcription expression, caspase-3 activity and p21 protein expression. Additionally, decreased levels of heat shock protein 27 (HSP27) phosphorylation at Ser78 was correlated with knockdown of ICT1 in U937 cells. Thus, we concluded that the regulatory role of ICT1 in leukemia may be used as a potential therapeutic target for the treatment of leukemia.
Original languageEnglish
Pages (from-to)1269-1275
Number of pages7
JournalOncology Reports
Volume39
Issue number3
Early online dateJan 2018
DOIs
Publication statusPublished - Mar 2018

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S Phase
Colon
Leukemia
Apoptosis
Carcinoma
U937 Cells
Cyclin A2
Far-Western Blotting
HSP27 Heat-Shock Proteins
Hepatoblastoma
Growth Inhibitors
Annexin A5
Glioblastoma
Ribosomes
Codon
Caspase 3
Small Interfering RNA
Real-Time Polymerase Chain Reaction
Coloring Agents
Down-Regulation

Cite this

Li, Guang Yao ; Liu, Ji Zhu ; Zhang, Li ; Liu, Guo Zhen ; Li, Shuang Jing ; Xiao, Tai Wu ; Wang, Jing Xia ; Wang, Le Xin ; Hou, Ming. / Knockdown of immature colon carcinoma transcript 1 induces suppression of proliferation, S-phase arrest and apoptosis in leukemia cells. In: Oncology Reports. 2018 ; Vol. 39, No. 3. pp. 1269-1275.
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abstract = "Immature colon carcinoma transcript 1 (ICT1), a human mitochondrial translation release factor, is a ribosome-dependent codon-independent peptidyl-tRNA hydrolase. ICT1-deficiency has been recognized as a cell growth inhibitor of hepatoblastoma and glioblastoma multiforme. To explore the role of ICT1 in human leukemia, 2 short hairpin RNAs (shRNAs) targeting ICT1 sequences were designed in leukemia U937 cells. The successful infection of ICT1 in the U937 cells was observed under a fluorescence microscope and further quantified by western blotting and quantitative real-time PCR (qRT-PCR) analysis. Tetrazolium dye (MTT) assay revealed a significant decrease in proliferation of ICT1-knockdown U937 cells on the fourth and fifth day as compared with the control. Depletion of ICT1 resulted in an increase in S phase and sub-G1 (representing cell apoptosis) fractions. Annexin V-APC/7-AAD staining assay confirmed that knockdown of ICT1 played a crucial role in boosting early and late apoptotic programs in U937 cells. Downregulation of ICT1 also altered cyclin A2 transcription expression, caspase-3 activity and p21 protein expression. Additionally, decreased levels of heat shock protein 27 (HSP27) phosphorylation at Ser78 was correlated with knockdown of ICT1 in U937 cells. Thus, we concluded that the regulatory role of ICT1 in leukemia may be used as a potential therapeutic target for the treatment of leukemia.",
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Knockdown of immature colon carcinoma transcript 1 induces suppression of proliferation, S-phase arrest and apoptosis in leukemia cells. / Li, Guang Yao; Liu, Ji Zhu; Zhang, Li; Liu, Guo Zhen; Li, Shuang Jing; Xiao, Tai Wu; Wang, Jing Xia; Wang, Le Xin; Hou, Ming.

In: Oncology Reports, Vol. 39, No. 3, 03.2018, p. 1269-1275.

Research output: Contribution to journalArticle

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T1 - Knockdown of immature colon carcinoma transcript 1 induces suppression of proliferation, S-phase arrest and apoptosis in leukemia cells

AU - Li, Guang Yao

AU - Liu, Ji Zhu

AU - Zhang, Li

AU - Liu, Guo Zhen

AU - Li, Shuang Jing

AU - Xiao, Tai Wu

AU - Wang, Jing Xia

AU - Wang, Le Xin

AU - Hou, Ming

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AB - Immature colon carcinoma transcript 1 (ICT1), a human mitochondrial translation release factor, is a ribosome-dependent codon-independent peptidyl-tRNA hydrolase. ICT1-deficiency has been recognized as a cell growth inhibitor of hepatoblastoma and glioblastoma multiforme. To explore the role of ICT1 in human leukemia, 2 short hairpin RNAs (shRNAs) targeting ICT1 sequences were designed in leukemia U937 cells. The successful infection of ICT1 in the U937 cells was observed under a fluorescence microscope and further quantified by western blotting and quantitative real-time PCR (qRT-PCR) analysis. Tetrazolium dye (MTT) assay revealed a significant decrease in proliferation of ICT1-knockdown U937 cells on the fourth and fifth day as compared with the control. Depletion of ICT1 resulted in an increase in S phase and sub-G1 (representing cell apoptosis) fractions. Annexin V-APC/7-AAD staining assay confirmed that knockdown of ICT1 played a crucial role in boosting early and late apoptotic programs in U937 cells. Downregulation of ICT1 also altered cyclin A2 transcription expression, caspase-3 activity and p21 protein expression. Additionally, decreased levels of heat shock protein 27 (HSP27) phosphorylation at Ser78 was correlated with knockdown of ICT1 in U937 cells. Thus, we concluded that the regulatory role of ICT1 in leukemia may be used as a potential therapeutic target for the treatment of leukemia.

KW - Apoptosis

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KW - Cell cycle

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