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Nrf2 is a potential prognostic marker and promotes proliferation and invasion in human hepatocellular carcinoma.

Zhang M, Zhang C, Zhang L, Yang Q, Zhou S, Wen Q, Wang J - BMC Cancer (2015)

Bottom Line: Further, the effect of Nrf2 on cell proliferation, apoptosis, and metastasis was examined in vitro by modulating expression of Nrf2 through specific shRNA or expression plasmid.Further studies demonstrated that inhibition of Nrf2 expression inhibited proliferation by inducing apoptosis and repressed invasion, and up-regulation of Nrf2 expression resulted in opposite phenotypes.Nrf2 is a potential prognostic marker and promotes proliferation and invasion in human hepatocellular carcinoma partly through regulating expression of Bcl-xL and MMP-9.

View Article: PubMed Central - PubMed

Affiliation: Department of Gastroenterology, Tangdu Hospital, Fourth Military Medical University, Xi'an, 710038, Shaanxi Province, China. zmx3115@163.com.

ABSTRACT

Background: Nuclear factor E2-related factor 2 (Nrf2 or NFE2L2) is abundantly expressed in cancer cells and relates to proliferation, invasion, and chemoresistance. Our early observations also found that expression of Nrf2 was up-regulated in kinds of cancer including human hepatocellular carcinoma (HCC) cells. But there are limited reports about the expression, significance, function of Nrf2 in HCC.

Methods: First, Nrf2 expression was analyzed in HCC cell lines and tumor samples. Then, the relationship of Nrf2 with clinicopathological factors and survival were analyzed. Further, the effect of Nrf2 on cell proliferation, apoptosis, and metastasis was examined in vitro by modulating expression of Nrf2 through specific shRNA or expression plasmid. Last, the potential mechanisms were also investigated.

Results: Nrf2 was up-regulated in HCC, and expression of Nrf2 was correlated with tumor differentiation, metastasis, and tumor size. Univariate and multivariate analyses indicated that high Nrf2 expression might be a poor prognostic factor. Further studies demonstrated that inhibition of Nrf2 expression inhibited proliferation by inducing apoptosis and repressed invasion, and up-regulation of Nrf2 expression resulted in opposite phenotypes. Moreover, there are positive correlation between Nrf2 expression and that of Bcl-xL and MMP-9.

Conclusion: Nrf2 is a potential prognostic marker and promotes proliferation and invasion in human hepatocellular carcinoma partly through regulating expression of Bcl-xL and MMP-9.

No MeSH data available.


Related in: MedlinePlus

Modulation of endogenous Nrf2 expression. a After transfected with Nrf2-shRNA (shRNA-867, shRNA-1118, shRNA-1757, or shRNA-2019) or control shRNA (shNC), expression levels of Nrf2 mRNA in Bel-7402 cells were detected by qRT-PCR; b-c After transfected with Nrf2-shRNA (shRNA-867, shRNA-1118, shRNA-1757, or shRNA-2019) or control shRNA (shNC), expression levels of Nrf2 protein in Bel-7402 cells were detected by western blot; d After transfected with Nrf2 expression plasmid (pEFGP-Nrf2-1 or pEFGP-Nrf2-2) or mock pEGFP plasmid (pEGFP-NC), expression levels of Nrf2 mRNA in HepG2 cells were detected by qRT-PCR; e-f After transfected with Nrf2 expression plasmid (pEFGP-Nrf2-1 or pEFGP-Nrf2-2) or mock pEGFP plasmid (pEGFP-NC), expression levels of Nrf2 protein in HepG2 cells were detected by western blot. *P <0.05 compared with control (Bel-7402 cells or HepG2 cells respectively) or shNC and pEGFP-NC
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Fig3: Modulation of endogenous Nrf2 expression. a After transfected with Nrf2-shRNA (shRNA-867, shRNA-1118, shRNA-1757, or shRNA-2019) or control shRNA (shNC), expression levels of Nrf2 mRNA in Bel-7402 cells were detected by qRT-PCR; b-c After transfected with Nrf2-shRNA (shRNA-867, shRNA-1118, shRNA-1757, or shRNA-2019) or control shRNA (shNC), expression levels of Nrf2 protein in Bel-7402 cells were detected by western blot; d After transfected with Nrf2 expression plasmid (pEFGP-Nrf2-1 or pEFGP-Nrf2-2) or mock pEGFP plasmid (pEGFP-NC), expression levels of Nrf2 mRNA in HepG2 cells were detected by qRT-PCR; e-f After transfected with Nrf2 expression plasmid (pEFGP-Nrf2-1 or pEFGP-Nrf2-2) or mock pEGFP plasmid (pEGFP-NC), expression levels of Nrf2 protein in HepG2 cells were detected by western blot. *P <0.05 compared with control (Bel-7402 cells or HepG2 cells respectively) or shNC and pEGFP-NC

Mentions: To knock down the endogenous expression of Nrf2 in Bel-7402 cells, we applied a plasmid vector expressing specific shRNA sequence targeting Nrf2 (shRNA-Nrf2). As a control, we stably transfected the Bel-7402 cells with the same plasmid vector expressing a control shRNA sequence (shNC) that did not target any known human gene. Through mRNA and protein expression analysis, we found that the shNC cells have a similar Nrf2 level as the parental Bel-7402 cells, which were significantly higher than the level in the shNrf2 cells (Fig. 3a, b and c). We then applied a expression plasmid named pEGFP-Nrf2 to up-regulate expression of Nrf2 in HepG2. The mRNA and protein expression analysis confirmed that pEGFP-Nrf2 significantly increased expression of Nrf2 in transfected HepG2 cells (Fig. 3d, e and f).Fig. 3


Nrf2 is a potential prognostic marker and promotes proliferation and invasion in human hepatocellular carcinoma.

Zhang M, Zhang C, Zhang L, Yang Q, Zhou S, Wen Q, Wang J - BMC Cancer (2015)

Modulation of endogenous Nrf2 expression. a After transfected with Nrf2-shRNA (shRNA-867, shRNA-1118, shRNA-1757, or shRNA-2019) or control shRNA (shNC), expression levels of Nrf2 mRNA in Bel-7402 cells were detected by qRT-PCR; b-c After transfected with Nrf2-shRNA (shRNA-867, shRNA-1118, shRNA-1757, or shRNA-2019) or control shRNA (shNC), expression levels of Nrf2 protein in Bel-7402 cells were detected by western blot; d After transfected with Nrf2 expression plasmid (pEFGP-Nrf2-1 or pEFGP-Nrf2-2) or mock pEGFP plasmid (pEGFP-NC), expression levels of Nrf2 mRNA in HepG2 cells were detected by qRT-PCR; e-f After transfected with Nrf2 expression plasmid (pEFGP-Nrf2-1 or pEFGP-Nrf2-2) or mock pEGFP plasmid (pEGFP-NC), expression levels of Nrf2 protein in HepG2 cells were detected by western blot. *P <0.05 compared with control (Bel-7402 cells or HepG2 cells respectively) or shNC and pEGFP-NC
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4507320&req=5

Fig3: Modulation of endogenous Nrf2 expression. a After transfected with Nrf2-shRNA (shRNA-867, shRNA-1118, shRNA-1757, or shRNA-2019) or control shRNA (shNC), expression levels of Nrf2 mRNA in Bel-7402 cells were detected by qRT-PCR; b-c After transfected with Nrf2-shRNA (shRNA-867, shRNA-1118, shRNA-1757, or shRNA-2019) or control shRNA (shNC), expression levels of Nrf2 protein in Bel-7402 cells were detected by western blot; d After transfected with Nrf2 expression plasmid (pEFGP-Nrf2-1 or pEFGP-Nrf2-2) or mock pEGFP plasmid (pEGFP-NC), expression levels of Nrf2 mRNA in HepG2 cells were detected by qRT-PCR; e-f After transfected with Nrf2 expression plasmid (pEFGP-Nrf2-1 or pEFGP-Nrf2-2) or mock pEGFP plasmid (pEGFP-NC), expression levels of Nrf2 protein in HepG2 cells were detected by western blot. *P <0.05 compared with control (Bel-7402 cells or HepG2 cells respectively) or shNC and pEGFP-NC
Mentions: To knock down the endogenous expression of Nrf2 in Bel-7402 cells, we applied a plasmid vector expressing specific shRNA sequence targeting Nrf2 (shRNA-Nrf2). As a control, we stably transfected the Bel-7402 cells with the same plasmid vector expressing a control shRNA sequence (shNC) that did not target any known human gene. Through mRNA and protein expression analysis, we found that the shNC cells have a similar Nrf2 level as the parental Bel-7402 cells, which were significantly higher than the level in the shNrf2 cells (Fig. 3a, b and c). We then applied a expression plasmid named pEGFP-Nrf2 to up-regulate expression of Nrf2 in HepG2. The mRNA and protein expression analysis confirmed that pEGFP-Nrf2 significantly increased expression of Nrf2 in transfected HepG2 cells (Fig. 3d, e and f).Fig. 3

Bottom Line: Further, the effect of Nrf2 on cell proliferation, apoptosis, and metastasis was examined in vitro by modulating expression of Nrf2 through specific shRNA or expression plasmid.Further studies demonstrated that inhibition of Nrf2 expression inhibited proliferation by inducing apoptosis and repressed invasion, and up-regulation of Nrf2 expression resulted in opposite phenotypes.Nrf2 is a potential prognostic marker and promotes proliferation and invasion in human hepatocellular carcinoma partly through regulating expression of Bcl-xL and MMP-9.

View Article: PubMed Central - PubMed

Affiliation: Department of Gastroenterology, Tangdu Hospital, Fourth Military Medical University, Xi'an, 710038, Shaanxi Province, China. zmx3115@163.com.

ABSTRACT

Background: Nuclear factor E2-related factor 2 (Nrf2 or NFE2L2) is abundantly expressed in cancer cells and relates to proliferation, invasion, and chemoresistance. Our early observations also found that expression of Nrf2 was up-regulated in kinds of cancer including human hepatocellular carcinoma (HCC) cells. But there are limited reports about the expression, significance, function of Nrf2 in HCC.

Methods: First, Nrf2 expression was analyzed in HCC cell lines and tumor samples. Then, the relationship of Nrf2 with clinicopathological factors and survival were analyzed. Further, the effect of Nrf2 on cell proliferation, apoptosis, and metastasis was examined in vitro by modulating expression of Nrf2 through specific shRNA or expression plasmid. Last, the potential mechanisms were also investigated.

Results: Nrf2 was up-regulated in HCC, and expression of Nrf2 was correlated with tumor differentiation, metastasis, and tumor size. Univariate and multivariate analyses indicated that high Nrf2 expression might be a poor prognostic factor. Further studies demonstrated that inhibition of Nrf2 expression inhibited proliferation by inducing apoptosis and repressed invasion, and up-regulation of Nrf2 expression resulted in opposite phenotypes. Moreover, there are positive correlation between Nrf2 expression and that of Bcl-xL and MMP-9.

Conclusion: Nrf2 is a potential prognostic marker and promotes proliferation and invasion in human hepatocellular carcinoma partly through regulating expression of Bcl-xL and MMP-9.

No MeSH data available.


Related in: MedlinePlus