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Hepatitis B virus induces G1 phase arrest by regulating cell cycle genes in HepG2.2.15 cells.

Wang T, Zhao R, Wu Y, Kong D, Zhang L, Wu D, Li C, Zhang C, Yu Z, Jin X - Virol. J. (2011)

Bottom Line: To investigate the effect of HBV on the proliferative ability of host cells and explore the potential mechanism.G1 phase arrest was the main cause but was not associated with apoptosis. p53, p21 and total retinoblastoma (Rb) were determined to be up-regulated, whereas cyclinE was down-regulated at both the protein and mRNA levels in HepG2.2.15 cells.The phosphorylated Rb in HepG2.2.15 cells was decreased.

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Affiliation: Department of Pathology, Basic Medical Science College, Harbin Medical University, 157 Baojian Road, Nangang District, Harbin 150081, China.

ABSTRACT

Background: To investigate the effect of HBV on the proliferative ability of host cells and explore the potential mechanism.

Methods: MTT, colony formation assay and tumourigenicity in nude mice were performed to investigate the effect of HBV on the proliferative capability of host cells. In order to explore the potential mechanism, cell cycle and apoptosis were analysed. The cell cycle genes controlling the G1/S phase transition were detected by immunohistochemistry, westernblot and RT-PCR.

Results: HepG2.2.15 cells showed decreased proliferation ability compared to HepG2 cells. G1 phase arrest was the main cause but was not associated with apoptosis. p53, p21 and total retinoblastoma (Rb) were determined to be up-regulated, whereas cyclinE was down-regulated at both the protein and mRNA levels in HepG2.2.15 cells. The phosphorylated Rb in HepG2.2.15 cells was decreased.

Conclusions: Our results suggested that HBV inhibited the capability of proliferation of HepG2.2.15 cells by regulating cell cycle genes expression and inducing G1 arrest.

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Protein and mRNA assays for the detection of p53, p21, p27, p16, cyclinD1, cyclinE, Rb and pRb. (A) Immunohistochemistry assay (400×). Bar, 50 μm. All proteins were located in nuclei and a positive result was brown nuclear staining. (B) Western blot analysis. (C) Semi-quantitative RT-PCR analysis. (D) The relative expression level of each protein was represented as the fold ratio. Compared with HepG2 cells. *p < 0.05, **p < 0.01.
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Figure 4: Protein and mRNA assays for the detection of p53, p21, p27, p16, cyclinD1, cyclinE, Rb and pRb. (A) Immunohistochemistry assay (400×). Bar, 50 μm. All proteins were located in nuclei and a positive result was brown nuclear staining. (B) Western blot analysis. (C) Semi-quantitative RT-PCR analysis. (D) The relative expression level of each protein was represented as the fold ratio. Compared with HepG2 cells. *p < 0.05, **p < 0.01.

Mentions: To explore the mechanism of G1 phase arrest, the levels of cell cycle regulator proteins controlling the G1/S phase transition were analyzed by western blot (Figure 4B). We determined that p53, p21 and total Rb were increased, while cyclinE and phosphorylated Rb were decreased in HepG2.2.15 cells compared to those in HepG2 cells. The levels of p16 and p27 showed no significant difference between the two cell lines. Immunohistochemical data confirmed that of the western blot (Figure 4A). CyclinD1 was increased in HepG2.2.15 by western blot, but there was no significant difference between the two cell lines by immunohistochemistry.


Hepatitis B virus induces G1 phase arrest by regulating cell cycle genes in HepG2.2.15 cells.

Wang T, Zhao R, Wu Y, Kong D, Zhang L, Wu D, Li C, Zhang C, Yu Z, Jin X - Virol. J. (2011)

Protein and mRNA assays for the detection of p53, p21, p27, p16, cyclinD1, cyclinE, Rb and pRb. (A) Immunohistochemistry assay (400×). Bar, 50 μm. All proteins were located in nuclei and a positive result was brown nuclear staining. (B) Western blot analysis. (C) Semi-quantitative RT-PCR analysis. (D) The relative expression level of each protein was represented as the fold ratio. Compared with HepG2 cells. *p < 0.05, **p < 0.01.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3104952&req=5

Figure 4: Protein and mRNA assays for the detection of p53, p21, p27, p16, cyclinD1, cyclinE, Rb and pRb. (A) Immunohistochemistry assay (400×). Bar, 50 μm. All proteins were located in nuclei and a positive result was brown nuclear staining. (B) Western blot analysis. (C) Semi-quantitative RT-PCR analysis. (D) The relative expression level of each protein was represented as the fold ratio. Compared with HepG2 cells. *p < 0.05, **p < 0.01.
Mentions: To explore the mechanism of G1 phase arrest, the levels of cell cycle regulator proteins controlling the G1/S phase transition were analyzed by western blot (Figure 4B). We determined that p53, p21 and total Rb were increased, while cyclinE and phosphorylated Rb were decreased in HepG2.2.15 cells compared to those in HepG2 cells. The levels of p16 and p27 showed no significant difference between the two cell lines. Immunohistochemical data confirmed that of the western blot (Figure 4A). CyclinD1 was increased in HepG2.2.15 by western blot, but there was no significant difference between the two cell lines by immunohistochemistry.

Bottom Line: To investigate the effect of HBV on the proliferative ability of host cells and explore the potential mechanism.G1 phase arrest was the main cause but was not associated with apoptosis. p53, p21 and total retinoblastoma (Rb) were determined to be up-regulated, whereas cyclinE was down-regulated at both the protein and mRNA levels in HepG2.2.15 cells.The phosphorylated Rb in HepG2.2.15 cells was decreased.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Pathology, Basic Medical Science College, Harbin Medical University, 157 Baojian Road, Nangang District, Harbin 150081, China.

ABSTRACT

Background: To investigate the effect of HBV on the proliferative ability of host cells and explore the potential mechanism.

Methods: MTT, colony formation assay and tumourigenicity in nude mice were performed to investigate the effect of HBV on the proliferative capability of host cells. In order to explore the potential mechanism, cell cycle and apoptosis were analysed. The cell cycle genes controlling the G1/S phase transition were detected by immunohistochemistry, westernblot and RT-PCR.

Results: HepG2.2.15 cells showed decreased proliferation ability compared to HepG2 cells. G1 phase arrest was the main cause but was not associated with apoptosis. p53, p21 and total retinoblastoma (Rb) were determined to be up-regulated, whereas cyclinE was down-regulated at both the protein and mRNA levels in HepG2.2.15 cells. The phosphorylated Rb in HepG2.2.15 cells was decreased.

Conclusions: Our results suggested that HBV inhibited the capability of proliferation of HepG2.2.15 cells by regulating cell cycle genes expression and inducing G1 arrest.

Show MeSH
Related in: MedlinePlus