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Identification of FN1BP1 as a novel cell cycle regulator through modulating G1 checkpoint in human hepatocarcinoma Hep3B cells.

Liu M, Wu R, Yang F, Wang T, Zhang P, Gu J, Wan D, Yang S - PLoS ONE (2013)

Bottom Line: Some interesting genes (p21cip1, ID2, GMSF, ERCC5, and RPA1), which changed in the cDNA microarray analysis, were confirmed by semi-qRT-PCR, and similar changes in expression were observed.FCM cell-cycle analysis indicated that FN1BP1 over-expression could result in G1 phase arrest.These results indicate the potential role of FN1BP1 as a treatment target for hepatocellular carcinoma.

View Article: PubMed Central - PubMed

Affiliation: Jingsu Key Laboratory of Neuroregeneration, Nantong University, Nantong, Jiangsu, China.

ABSTRACT
A novel human gene, FN1BP1 (fibronectin 1 binding protein 1), was identified using the human placenta cDNA library. Northern blotting showed a transcript of ∼2.8 kb in human placenta, liver, and skeletal muscle tissues. This mRNA transcript length was similar to the full FN1BP1 sequence obtained previously. We established a conditionally induced stable cell line of Hep3B-Tet-on-FN1BP1 to investigate the preliminary function and mechanism of the secretory FN1BP1 protein. Cell-proliferation and colony-conformation assays demonstrated that FN1BP1 protein suppressed Hep3B cell growth and colonization in vitro. Analysis of Atlas human cDNA expression indicated that after FN1BP1 Dox-inducing expression for 24 h, 19 genes were up-regulated and 22 genes were down-regulated more than two-fold. Most of these gene changes were related to cell-cycle-arrest proteins (p21cip1, p15, and cyclin E1), transcription factors (general transcription factors, zinc finger proteins, transcriptional enhancer factors), SWI/SNF (SWItch/Sucrose NonFermentable) complex units, early-response proteins, and nerve growth or neurotrophic factors. Down-regulated genes were subject to colony-stimulating factors (e.g., GMSFs), and many repair genes were involved in DNA damage (RAD, ERCC, DNA topoisomerase, polymerase, and ligase). Some interesting genes (p21cip1, ID2, GMSF, ERCC5, and RPA1), which changed in the cDNA microarray analysis, were confirmed by semi-qRT-PCR, and similar changes in expression were observed. FCM cell-cycle analysis indicated that FN1BP1 over-expression could result in G1 phase arrest. FN1BP1 might inhibit cell growth and/or colony conformation through G1 phase arrest of the Hep3B cell cycle. These results indicate the potential role of FN1BP1 as a treatment target for hepatocellular carcinoma.

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FN1BP1 expression in human placenta, liver, and skeletal muscle tissues.The tissue expression pattern of FN1BP1 (Accession No. AF217970) was investigated by hybridizing its cDNA probe to a multiple-tissue northern blot assay. The mRNA content in the eight lanes are from heart, brain, placenta, lung, liver, skeletal muscle, kidney, and pancreas tissues, respectively. A transcript of ∼2.8 kb was observed in human placenta, liver, and skeletal muscle tissues. The arrow (no tail) shows this ∼2.8-kb length of the mRNA transcript. The arrows at 2.4 kb and 4.4 kb indicate the mean sizes of the RNA markers.
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pone-0057574-g001: FN1BP1 expression in human placenta, liver, and skeletal muscle tissues.The tissue expression pattern of FN1BP1 (Accession No. AF217970) was investigated by hybridizing its cDNA probe to a multiple-tissue northern blot assay. The mRNA content in the eight lanes are from heart, brain, placenta, lung, liver, skeletal muscle, kidney, and pancreas tissues, respectively. A transcript of ∼2.8 kb was observed in human placenta, liver, and skeletal muscle tissues. The arrow (no tail) shows this ∼2.8-kb length of the mRNA transcript. The arrows at 2.4 kb and 4.4 kb indicate the mean sizes of the RNA markers.

Mentions: The tissue expression pattern of FN1BP1 was investigated using multiple-tissue northern blotting. Figure 1 shows the result of northern blotting for human MTN using the FN1BP1 probe. A ∼2.8-kb fragment was observed in human placenta, liver and skeletal muscle tissues. This mRNA transcript length was similar to the full FN1BP1 sequence we obtained previously.


Identification of FN1BP1 as a novel cell cycle regulator through modulating G1 checkpoint in human hepatocarcinoma Hep3B cells.

Liu M, Wu R, Yang F, Wang T, Zhang P, Gu J, Wan D, Yang S - PLoS ONE (2013)

FN1BP1 expression in human placenta, liver, and skeletal muscle tissues.The tissue expression pattern of FN1BP1 (Accession No. AF217970) was investigated by hybridizing its cDNA probe to a multiple-tissue northern blot assay. The mRNA content in the eight lanes are from heart, brain, placenta, lung, liver, skeletal muscle, kidney, and pancreas tissues, respectively. A transcript of ∼2.8 kb was observed in human placenta, liver, and skeletal muscle tissues. The arrow (no tail) shows this ∼2.8-kb length of the mRNA transcript. The arrows at 2.4 kb and 4.4 kb indicate the mean sizes of the RNA markers.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3585200&req=5

pone-0057574-g001: FN1BP1 expression in human placenta, liver, and skeletal muscle tissues.The tissue expression pattern of FN1BP1 (Accession No. AF217970) was investigated by hybridizing its cDNA probe to a multiple-tissue northern blot assay. The mRNA content in the eight lanes are from heart, brain, placenta, lung, liver, skeletal muscle, kidney, and pancreas tissues, respectively. A transcript of ∼2.8 kb was observed in human placenta, liver, and skeletal muscle tissues. The arrow (no tail) shows this ∼2.8-kb length of the mRNA transcript. The arrows at 2.4 kb and 4.4 kb indicate the mean sizes of the RNA markers.
Mentions: The tissue expression pattern of FN1BP1 was investigated using multiple-tissue northern blotting. Figure 1 shows the result of northern blotting for human MTN using the FN1BP1 probe. A ∼2.8-kb fragment was observed in human placenta, liver and skeletal muscle tissues. This mRNA transcript length was similar to the full FN1BP1 sequence we obtained previously.

Bottom Line: Some interesting genes (p21cip1, ID2, GMSF, ERCC5, and RPA1), which changed in the cDNA microarray analysis, were confirmed by semi-qRT-PCR, and similar changes in expression were observed.FCM cell-cycle analysis indicated that FN1BP1 over-expression could result in G1 phase arrest.These results indicate the potential role of FN1BP1 as a treatment target for hepatocellular carcinoma.

View Article: PubMed Central - PubMed

Affiliation: Jingsu Key Laboratory of Neuroregeneration, Nantong University, Nantong, Jiangsu, China.

ABSTRACT
A novel human gene, FN1BP1 (fibronectin 1 binding protein 1), was identified using the human placenta cDNA library. Northern blotting showed a transcript of ∼2.8 kb in human placenta, liver, and skeletal muscle tissues. This mRNA transcript length was similar to the full FN1BP1 sequence obtained previously. We established a conditionally induced stable cell line of Hep3B-Tet-on-FN1BP1 to investigate the preliminary function and mechanism of the secretory FN1BP1 protein. Cell-proliferation and colony-conformation assays demonstrated that FN1BP1 protein suppressed Hep3B cell growth and colonization in vitro. Analysis of Atlas human cDNA expression indicated that after FN1BP1 Dox-inducing expression for 24 h, 19 genes were up-regulated and 22 genes were down-regulated more than two-fold. Most of these gene changes were related to cell-cycle-arrest proteins (p21cip1, p15, and cyclin E1), transcription factors (general transcription factors, zinc finger proteins, transcriptional enhancer factors), SWI/SNF (SWItch/Sucrose NonFermentable) complex units, early-response proteins, and nerve growth or neurotrophic factors. Down-regulated genes were subject to colony-stimulating factors (e.g., GMSFs), and many repair genes were involved in DNA damage (RAD, ERCC, DNA topoisomerase, polymerase, and ligase). Some interesting genes (p21cip1, ID2, GMSF, ERCC5, and RPA1), which changed in the cDNA microarray analysis, were confirmed by semi-qRT-PCR, and similar changes in expression were observed. FCM cell-cycle analysis indicated that FN1BP1 over-expression could result in G1 phase arrest. FN1BP1 might inhibit cell growth and/or colony conformation through G1 phase arrest of the Hep3B cell cycle. These results indicate the potential role of FN1BP1 as a treatment target for hepatocellular carcinoma.

Show MeSH
Related in: MedlinePlus