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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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The semi-quantitative RT-PCR analysis of p21cip1, ID2, GMSF, ERCC5 and RPA1.(A) The mRNA levels of five genes, p21cip1, ID2, GMSF, ERCC5, and RPA1 were selected for evaluation using semi-quantitative RT-PCR. The mRNA expression of those target genes was consistent with the hybridization data for each of the genes measured. That is, mRNA levels of p21cip1 and ID2 were up-regulated (upper panel), while mRNA levels of RPA1, ERCC5, and GMSF were down-regulated in FN1BP1 over-expressed cells (lower panel). The statistical result of the semi-quantitative RT-PCR of the selected genes is shown in (B).
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pone-0057574-g004: The semi-quantitative RT-PCR analysis of p21cip1, ID2, GMSF, ERCC5 and RPA1.(A) The mRNA levels of five genes, p21cip1, ID2, GMSF, ERCC5, and RPA1 were selected for evaluation using semi-quantitative RT-PCR. The mRNA expression of those target genes was consistent with the hybridization data for each of the genes measured. That is, mRNA levels of p21cip1 and ID2 were up-regulated (upper panel), while mRNA levels of RPA1, ERCC5, and GMSF were down-regulated in FN1BP1 over-expressed cells (lower panel). The statistical result of the semi-quantitative RT-PCR of the selected genes is shown in (B).

Mentions: Alteration in gene expression on Hep3B Tet-On FN1BP1/S11 cells was assessed after Dox induction for 24 h. The data show that, compared with non-Dox Heb3B cells, 19 genes were up-regulated (Table 2) and 22 genes were down-regulated (Table 3) more than twofold in Dox-induced FN1BP1 expressing Hep3B cells. Of these gene changes and their putative functions, which were up-regulated compared with those of the non- Dox-induced group, most were cell-cycle–arrest proteins (p21cip1, p15, and cyclin E1), transcription factors (general transcription factors, zinc finger proteins, and transcriptional enhancer factors), SWItch/Sucrose NonFermentable (SWI/SNF) complex units, early-response proteins, and nerve growth or neurotrophic factors. On the other hand, down-regulated genes were subject to colony-stimulating factors (e.g., GMSF) and receptors, many repair genes after DNA damage (e.g., RAD, ERCC, DNA topoisomerase, polymerase, and ligase). Some genes (e.g., p21cip1, ID2, GMSF, ERCC5, and RPA), which that changed in the cDNA microarray analysis, were confirmed by semi-qRT-PCR, and similar changes in expression were observed (Fig. 4).


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)

The semi-quantitative RT-PCR analysis of p21cip1, ID2, GMSF, ERCC5 and RPA1.(A) The mRNA levels of five genes, p21cip1, ID2, GMSF, ERCC5, and RPA1 were selected for evaluation using semi-quantitative RT-PCR. The mRNA expression of those target genes was consistent with the hybridization data for each of the genes measured. That is, mRNA levels of p21cip1 and ID2 were up-regulated (upper panel), while mRNA levels of RPA1, ERCC5, and GMSF were down-regulated in FN1BP1 over-expressed cells (lower panel). The statistical result of the semi-quantitative RT-PCR of the selected genes is shown in (B).
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Related In: Results  -  Collection

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

pone-0057574-g004: The semi-quantitative RT-PCR analysis of p21cip1, ID2, GMSF, ERCC5 and RPA1.(A) The mRNA levels of five genes, p21cip1, ID2, GMSF, ERCC5, and RPA1 were selected for evaluation using semi-quantitative RT-PCR. The mRNA expression of those target genes was consistent with the hybridization data for each of the genes measured. That is, mRNA levels of p21cip1 and ID2 were up-regulated (upper panel), while mRNA levels of RPA1, ERCC5, and GMSF were down-regulated in FN1BP1 over-expressed cells (lower panel). The statistical result of the semi-quantitative RT-PCR of the selected genes is shown in (B).
Mentions: Alteration in gene expression on Hep3B Tet-On FN1BP1/S11 cells was assessed after Dox induction for 24 h. The data show that, compared with non-Dox Heb3B cells, 19 genes were up-regulated (Table 2) and 22 genes were down-regulated (Table 3) more than twofold in Dox-induced FN1BP1 expressing Hep3B cells. Of these gene changes and their putative functions, which were up-regulated compared with those of the non- Dox-induced group, most were cell-cycle–arrest proteins (p21cip1, p15, and cyclin E1), transcription factors (general transcription factors, zinc finger proteins, and transcriptional enhancer factors), SWItch/Sucrose NonFermentable (SWI/SNF) complex units, early-response proteins, and nerve growth or neurotrophic factors. On the other hand, down-regulated genes were subject to colony-stimulating factors (e.g., GMSF) and receptors, many repair genes after DNA damage (e.g., RAD, ERCC, DNA topoisomerase, polymerase, and ligase). Some genes (e.g., p21cip1, ID2, GMSF, ERCC5, and RPA), which that changed in the cDNA microarray analysis, were confirmed by semi-qRT-PCR, and similar changes in expression were observed (Fig. 4).

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