Effects of 9‐cis retinoic acid on human homeobox gene NKX3.1 expression in prostate cancer cell line LNCaP

Aim: To study the regulatory effects of 9‐cis retinoic acid (RA) on the expression of human homeobox gene NKX3.1 in prostate cancer cell line LNCaP. Methods: Flow cytometry, reverse transcriptase polymerase chain reaction and Western blotting were performed to evaluate the effects of 9‐cis RA on NKX...

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Published inAsian journal of andrology Vol. 8; no. 4; pp. 435 - 441
Main Authors Jiang, An‐Li, Zhang, Peng‐Ju, Chen, Wei‐Wen, Liu, Wen‐Wen, Yu, Chun‐Xiao, Hu, Xiao‐Yan, Zhang, Xiao‐Qian, Zhang, Jian‐Ye
Format Journal Article
LanguageEnglish
Published Melbourne, Australia Blackwell Publishing Asia 01.07.2006
Medknow Publications & Media Pvt. Ltd
Subjects
9‐cis retinoic acid
Base Sequence
Cell Cycle
Cell Differentiation
Cell Line, Tumor
DNA Primers
Flow Cytometry
gene expression
Gene Expression Regulation, Neoplastic - drug effects
Homeodomain Proteins - genetics
Humans
Male
NKX3.1 gene
Promoter Regions, Genetic
prostate cancer cell
Prostatic Neoplasms - genetics
Prostatic Neoplasms - pathology
Reverse Transcriptase Polymerase Chain Reaction
Transcription Factors - genetics
Tretinoin - pharmacology
Up-Regulation
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Abstract Aim: To study the regulatory effects of 9‐cis retinoic acid (RA) on the expression of human homeobox gene NKX3.1 in prostate cancer cell line LNCaP. Methods: Flow cytometry, reverse transcriptase polymerase chain reaction and Western blotting were performed to evaluate the effects of 9‐cis RA on NKX3.1 expression and cell cycle of LNCaP cells. To identify a regulatory region within the NKX3.1 promoter contributing to the regulation induced by 9‐cis RA, we have constructed an NKX3.1 promoter‐reporter plasmid, pGL3‐1040bp, and its 5′‐deletion mutants, which were transfected into LNCaP cells with treatment of 9‐cis RA in indicated concentrations. Results: With the treatment of 9‐cis RA, the NKX3.1 promoter activity was increased in reporter gene assay and NKX3.1 expression was enhanced at both mRNA and protein levels in LNCaP cells. We found that the region between ‐936 and ‐921 in the upstream of NKX3.1 gene involved the inducible regulation by 9‐cis RA treatment. In flow cytometry, 9‐cis RA treatment caused accumulation of cells in the G1 phase of the cell cycle and a fewer cells pass through to G2/M. Conclusion: Our results demonstrated that 9‐cis RA as a differentiating agent can arrest prostate cancer cells in G1 phase and reduce cell mitosis, and upregulate the expression of human homeobox gene NKX3.1, which is thought to play an important role in prostate differentiation and to act as a tumor suppressor gene in the prostate.
AbstractList Aim: To study the regulatory effects of 9‐cis retinoic acid (RA) on the expression of human homeobox gene NKX3.1 in prostate cancer cell line LNCaP. Methods: Flow cytometry, reverse transcriptase polymerase chain reaction and Western blotting were performed to evaluate the effects of 9‐cis RA on NKX3.1 expression and cell cycle of LNCaP cells. To identify a regulatory region within the NKX3.1 promoter contributing to the regulation induced by 9‐cis RA, we have constructed an NKX3.1 promoter‐reporter plasmid, pGL3‐1040bp, and its 5′‐deletion mutants, which were transfected into LNCaP cells with treatment of 9‐cis RA in indicated concentrations. Results: With the treatment of 9‐cis RA, the NKX3.1 promoter activity was increased in reporter gene assay and NKX3.1 expression was enhanced at both mRNA and protein levels in LNCaP cells. We found that the region between ‐936 and ‐921 in the upstream of NKX3.1 gene involved the inducible regulation by 9‐cis RA treatment. In flow cytometry, 9‐cis RA treatment caused accumulation of cells in the G1 phase of the cell cycle and a fewer cells pass through to G2/M. Conclusion: Our results demonstrated that 9‐cis RA as a differentiating agent can arrest prostate cancer cells in G1 phase and reduce cell mitosis, and upregulate the expression of human homeobox gene NKX3.1, which is thought to play an important role in prostate differentiation and to act as a tumor suppressor gene in the prostate.
To study the regulatory effects of 9-cis retinoic acid (RA) on the expression of human homeobox gene NKX3.1 in prostate cancer cell line LNCaP. Flow cytometry, reverse transcriptase polymerase chain reaction and Western blotting were performed to evaluate the effects of 9-cis RA on NKX3.1 expression and cell cycle of LNCaP cells. To identify a regulatory region within the NKX3.1 promoter contributing to the regulation induced by 9-cis RA, we have constructed an NKX3.1 promoter-reporter plasmid, pGL3-1040bp, and its 5'-deletion mutants, which were transfected into LNCaP cells with treatment of 9-cis RA in indicated concentrations. With the treatment of 9-cis RA, the NKX3.1 promoter activity was increased in reporter gene assay and NKX3.1 expression was enhanced at both mRNA and protein levels in LNCaP cells. We found that the region between -936 and -921 in the upstream of NKX3.1 gene involved the inducible regulation by 9-cis RA treatment. In flow cytometry, 9-cis RA treatment caused accumulation of cells in the G(1) phase of the cell cycle and a fewer cells pass through to G(2)/M. Our results demonstrated that 9-cis RA as a differentiating agent can arrest prostate cancer cells in G(1) phase and reduce cell mitosis, and upregulate the expression of human homeobox gene NKX3.1, which is thought to play an important role in prostate differentiation and to act as a tumor suppressor gene in the prostate.
Aim: To study the regulatory effects of 9-cis retinoic acid (RA) on the expression of human homeobox gene NKX3.1 in prostate cancer cell line LNCaP. Methods: Flow cytometry, reverse transcriptase polymerase chain reaction and Western blotting were performed to evaluate the effects of 9-cis RA on NKX3.1 expression and cell cycle of LNCaP cells. To identify a regulatory region within the NKX3.1 promoter contributing to the regulation induced by 9-cis RA, we have constructed an NKX3.1 promoter-reporter plasmid, pGL sub(3)-1040bp, and its 5'-deletion mutants, which were transfected into LNCaP cells with treatment of 9-cis RA in indicated concentrations. Results: With the treatment of 9-cis RA, the NKX3.1 promoter activity was increased in reporter gene assay and NKX3.1 expression was enhanced at both mRNA and protein levels in LNCaP cells. We found that the region between -936 and -921 in the upstream of NKX3.1 gene involved the inducible regulation by 9-cis RA treatment. In flow cytometry, 9-cis RA treatment caused accumulation of cells in the G sub(1) phase of the cell cycle and a fewer cells pass through to G sub(2)-M. Conclusion: Our results demonstrated that 9-cis RA as a differentiating agent can arrest prostate cancer cells in G sub(1) phase and reduce cell mitosis, and upregulate the expression of human homeobox gene NKX3.1, which is thought to play an important role in prostate differentiation and to act as a tumor suppressor gene in the prostate.
To study the regulatory effects of 9-cis retinoic acid (RA) on the expression of human homeobox gene NKX3.1 in prostate cancer cell line LNCaP. Flow cytometry, reverse transcriptase polymerase chain reaction and Western blotting were performed to evaluate the effects of 9-cis RA on NKX3.1 expression and cell cycle of LNCaP cells. To identify a regulatory region within the NKX3.1 promoter contributing to the regulation induced by 9-cis RA, we have constructed an NKX3.1 promoter-reporter plasmid, pGL3-1040bp, and its 5'-deletion mutants, which were transfected into LNCaP cells with treatment of 9-cis RA in indicated concentrations. With the treatment of 9-cis RA, the NKX3.1 promoter activity was increased in reporter gene assay and NKX3.1 expression was enhanced at both mRNA and protein levels in LNCaP cells. We found that the region between -936 and -921 in the upstream of NKX3.1 gene involved the inducible regulation by 9-cis RA treatment. In flow cytometry, 9-cis RA treatment caused accumulation of cells in the G(1) phase of the cell cycle and a fewer cells pass through to G(2)/M. Our results demonstrated that 9-cis RA as a differentiating agent can arrest prostate cancer cells in G(1) phase and reduce cell mitosis, and upregulate the expression of human homeobox gene NKX3.1, which is thought to play an important role in prostate differentiation and to act as a tumor suppressor gene in the prostate.
To study the regulatory effects of 9-cis retinoic acid (RA) on the expression of human homeobox gene NKX3.1 in prostate cancer cell line LNCaP.AIMTo study the regulatory effects of 9-cis retinoic acid (RA) on the expression of human homeobox gene NKX3.1 in prostate cancer cell line LNCaP.Flow cytometry, reverse transcriptase polymerase chain reaction and Western blotting were performed to evaluate the effects of 9-cis RA on NKX3.1 expression and cell cycle of LNCaP cells. To identify a regulatory region within the NKX3.1 promoter contributing to the regulation induced by 9-cis RA, we have constructed an NKX3.1 promoter-reporter plasmid, pGL3-1040bp, and its 5'-deletion mutants, which were transfected into LNCaP cells with treatment of 9-cis RA in indicated concentrations.METHODSFlow cytometry, reverse transcriptase polymerase chain reaction and Western blotting were performed to evaluate the effects of 9-cis RA on NKX3.1 expression and cell cycle of LNCaP cells. To identify a regulatory region within the NKX3.1 promoter contributing to the regulation induced by 9-cis RA, we have constructed an NKX3.1 promoter-reporter plasmid, pGL3-1040bp, and its 5'-deletion mutants, which were transfected into LNCaP cells with treatment of 9-cis RA in indicated concentrations.With the treatment of 9-cis RA, the NKX3.1 promoter activity was increased in reporter gene assay and NKX3.1 expression was enhanced at both mRNA and protein levels in LNCaP cells. We found that the region between -936 and -921 in the upstream of NKX3.1 gene involved the inducible regulation by 9-cis RA treatment. In flow cytometry, 9-cis RA treatment caused accumulation of cells in the G(1) phase of the cell cycle and a fewer cells pass through to G(2)/M.RESULTSWith the treatment of 9-cis RA, the NKX3.1 promoter activity was increased in reporter gene assay and NKX3.1 expression was enhanced at both mRNA and protein levels in LNCaP cells. We found that the region between -936 and -921 in the upstream of NKX3.1 gene involved the inducible regulation by 9-cis RA treatment. In flow cytometry, 9-cis RA treatment caused accumulation of cells in the G(1) phase of the cell cycle and a fewer cells pass through to G(2)/M.Our results demonstrated that 9-cis RA as a differentiating agent can arrest prostate cancer cells in G(1) phase and reduce cell mitosis, and upregulate the expression of human homeobox gene NKX3.1, which is thought to play an important role in prostate differentiation and to act as a tumor suppressor gene in the prostate.CONCLUSIONOur results demonstrated that 9-cis RA as a differentiating agent can arrest prostate cancer cells in G(1) phase and reduce cell mitosis, and upregulate the expression of human homeobox gene NKX3.1, which is thought to play an important role in prostate differentiation and to act as a tumor suppressor gene in the prostate.
Author Hu, Xiao‐Yan
Zhang, Xiao‐Qian
Chen, Wei‐Wen
Zhang, Peng‐Ju
Jiang, An‐Li
Yu, Chun‐Xiao
Zhang, Jian‐Ye
Liu, Wen‐Wen
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Snippet Aim: To study the regulatory effects of 9‐cis retinoic acid (RA) on the expression of human homeobox gene NKX3.1 in prostate cancer cell line LNCaP. Methods:...
To study the regulatory effects of 9-cis retinoic acid (RA) on the expression of human homeobox gene NKX3.1 in prostate cancer cell line LNCaP. Flow cytometry,...
To study the regulatory effects of 9-cis retinoic acid (RA) on the expression of human homeobox gene NKX3.1 in prostate cancer cell line LNCaP. Flow cytometry,...
Aim: To study the regulatory effects of 9-cis retinoic acid (RA) on the expression of human homeobox gene NKX3.1 in prostate cancer cell line LNCaP. Methods:...
To study the regulatory effects of 9-cis retinoic acid (RA) on the expression of human homeobox gene NKX3.1 in prostate cancer cell line LNCaP.AIMTo study the...
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StartPage 435
SubjectTerms 9‐cis retinoic acid
Base Sequence
Cell Cycle
Cell Differentiation
Cell Line, Tumor
DNA Primers
Flow Cytometry
gene expression
Gene Expression Regulation, Neoplastic - drug effects
Homeodomain Proteins - genetics
Humans
Male
NKX3.1 gene
Promoter Regions, Genetic
prostate cancer cell
Prostatic Neoplasms - genetics
Prostatic Neoplasms - pathology
Reverse Transcriptase Polymerase Chain Reaction
Transcription Factors - genetics
Tretinoin - pharmacology
Up-Regulation
Title Effects of 9‐cis retinoic acid on human homeobox gene NKX3.1 expression in prostate cancer cell line LNCaP
URI https://onlinelibrary.wiley.com/doi/abs/10.1111%2Fj.1745-7262.2006.00171.x
https://www.ncbi.nlm.nih.gov/pubmed/16763719
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https://www.proquest.com/docview/68045290
Volume 8
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