Enhancement of NIH3T3 cell proliferation by expressing macrophage colony stimulating factor in nuclei

Zhen Yu Cao; Ke Fu Wu; Ge Li; Yong Min Lin; Bin Zhang; Guo Guang Zheng

doi:10.1007/s11670-003-0010-6

Enhancement of NIH3T3 cell proliferation by expressing macrophage colony stimulating factor in nuclei

Zhen Yu Cao, Ke Fu Wu^*, Ge Li, Yong Min Lin, Bin Zhang, Guo Guang Zheng

^*Corresponding author for this work

Medicine, Hematology Oncology Division

Research output: Contribution to journal › Article › peer-review

2 Scopus citations

Abstract

Objective: To explore the effects of nuclear M-CSF on the process of tumorigenesis. Methods: Functional part of M-CSF cDNA was inserted into an eukaryotic expression plasmid pCMV/myc/nuc, which can add three NLS to the C-terminal of the expressed protein and direct the protein into the cell nuclei. The constructed plasmid was transferred into NIH3T3 cells and the cell clones were selected by G-418 selection. Cell clones stable expressing target protein were identified by RT-PCR, ABC immunohistochemistry assay and Western blot. Cell growth kinetics analyses through growth curves, cell doubling time, MTT test and anti-sense oligodeoxynucleotide (ASODN) inhibiting cell growth test were performed to identify cells proliferation potential. Results: The transfected cells showed elevated proliferation potential over the control cells. Conclusion: Abnormal appearance of M-CSF in nucleus could enhance cell proliferation, which suggests that cytokine isoforms within cell nucleus might play transcription factor-like role.

Original language	English (US)
Pages (from-to)	43-47
Number of pages	5
Journal	Chinese Journal of Cancer Research
Volume	15
Issue number	1
DOIs	https://doi.org/10.1007/s11670-003-0010-6
State	Published - Mar 2003

Keywords

Eukaryotic expression
Macrophage colony stimulating factor (M-CSF)
NIH3T3
Nuclear localization sequence (NLS)
Tumorigenesis

ASJC Scopus subject areas

Oncology
Cancer Research

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.1007/s11670-003-0010-6

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title = "Enhancement of NIH3T3 cell proliferation by expressing macrophage colony stimulating factor in nuclei",

abstract = "Objective: To explore the effects of nuclear M-CSF on the process of tumorigenesis. Methods: Functional part of M-CSF cDNA was inserted into an eukaryotic expression plasmid pCMV/myc/nuc, which can add three NLS to the C-terminal of the expressed protein and direct the protein into the cell nuclei. The constructed plasmid was transferred into NIH3T3 cells and the cell clones were selected by G-418 selection. Cell clones stable expressing target protein were identified by RT-PCR, ABC immunohistochemistry assay and Western blot. Cell growth kinetics analyses through growth curves, cell doubling time, MTT test and anti-sense oligodeoxynucleotide (ASODN) inhibiting cell growth test were performed to identify cells proliferation potential. Results: The transfected cells showed elevated proliferation potential over the control cells. Conclusion: Abnormal appearance of M-CSF in nucleus could enhance cell proliferation, which suggests that cytokine isoforms within cell nucleus might play transcription factor-like role.",

keywords = "Eukaryotic expression, Macrophage colony stimulating factor (M-CSF), NIH3T3, Nuclear localization sequence (NLS), Tumorigenesis",

author = "Cao, {Zhen Yu} and Wu, {Ke Fu} and Ge Li and Lin, {Yong Min} and Bin Zhang and Zheng, {Guo Guang}",

note = "Funding Information: Received date: November 18, 2002; Accepted date: January 17, 2003 Foundation item: This work was supported by a grant from Tianjin Science and Technology Development Project (No. 003119311). *Author to whom correspondence should be addressed. Phone: (0086-22)-27230400; Fax: (0086-22)-27306542; E-mail: kfwu @p ublic.tpt.tj.cn Biography: CAO Zhen-yu (1976-), male, graduated student, Institute of Hematology & Blood Disease Hospital, Chinese Academy of Medical Sciences & Peking Union Medical College, majors in cell biology.",

year = "2003",

month = mar,

doi = "10.1007/s11670-003-0010-6",

language = "English (US)",

volume = "15",

pages = "43--47",

journal = "Chinese Journal of Cancer Research",

issn = "1000-9604",

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T1 - Enhancement of NIH3T3 cell proliferation by expressing macrophage colony stimulating factor in nuclei

AU - Cao, Zhen Yu

AU - Wu, Ke Fu

AU - Li, Ge

AU - Lin, Yong Min

AU - Zhang, Bin

AU - Zheng, Guo Guang

N1 - Funding Information: Received date: November 18, 2002; Accepted date: January 17, 2003 Foundation item: This work was supported by a grant from Tianjin Science and Technology Development Project (No. 003119311). *Author to whom correspondence should be addressed. Phone: (0086-22)-27230400; Fax: (0086-22)-27306542; E-mail: kfwu @p ublic.tpt.tj.cn Biography: CAO Zhen-yu (1976-), male, graduated student, Institute of Hematology & Blood Disease Hospital, Chinese Academy of Medical Sciences & Peking Union Medical College, majors in cell biology.

PY - 2003/3

Y1 - 2003/3

N2 - Objective: To explore the effects of nuclear M-CSF on the process of tumorigenesis. Methods: Functional part of M-CSF cDNA was inserted into an eukaryotic expression plasmid pCMV/myc/nuc, which can add three NLS to the C-terminal of the expressed protein and direct the protein into the cell nuclei. The constructed plasmid was transferred into NIH3T3 cells and the cell clones were selected by G-418 selection. Cell clones stable expressing target protein were identified by RT-PCR, ABC immunohistochemistry assay and Western blot. Cell growth kinetics analyses through growth curves, cell doubling time, MTT test and anti-sense oligodeoxynucleotide (ASODN) inhibiting cell growth test were performed to identify cells proliferation potential. Results: The transfected cells showed elevated proliferation potential over the control cells. Conclusion: Abnormal appearance of M-CSF in nucleus could enhance cell proliferation, which suggests that cytokine isoforms within cell nucleus might play transcription factor-like role.

AB - Objective: To explore the effects of nuclear M-CSF on the process of tumorigenesis. Methods: Functional part of M-CSF cDNA was inserted into an eukaryotic expression plasmid pCMV/myc/nuc, which can add three NLS to the C-terminal of the expressed protein and direct the protein into the cell nuclei. The constructed plasmid was transferred into NIH3T3 cells and the cell clones were selected by G-418 selection. Cell clones stable expressing target protein were identified by RT-PCR, ABC immunohistochemistry assay and Western blot. Cell growth kinetics analyses through growth curves, cell doubling time, MTT test and anti-sense oligodeoxynucleotide (ASODN) inhibiting cell growth test were performed to identify cells proliferation potential. Results: The transfected cells showed elevated proliferation potential over the control cells. Conclusion: Abnormal appearance of M-CSF in nucleus could enhance cell proliferation, which suggests that cytokine isoforms within cell nucleus might play transcription factor-like role.

KW - Eukaryotic expression

KW - Macrophage colony stimulating factor (M-CSF)

KW - NIH3T3

KW - Nuclear localization sequence (NLS)

KW - Tumorigenesis

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Enhancement of NIH3T3 cell proliferation by expressing macrophage colony stimulating factor in nuclei

Abstract

Keywords

ASJC Scopus subject areas

UN SDGs

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