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Targeting protein arginine methyltransferase 5 inhibits human hepatocellular carcinoma growth via the downregulation of beta-catenin.

Zhang B, Dong S, Li Z, Lu L, Zhang S, Chen X, Cen X, Wu Y - J Transl Med (2015)

Bottom Line: Furthermore, β-catenin was identified as a target of PRMT5.Silencing PRMT5 significantly down-regulated the expression of β-catenin and the downstream effector Cyclin D1 in HCC cells.Meanwhile, AMI-1 decreased the expression levels of symmetric dimethylation of H4 (H4R3me2s), a histone mark of PRMT5.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacology, School of Basic Medical Sciences, Lanzhou University; Key Lab of Preclinical Study for New Drugs of Gansu Province, No 199, Dongang West Road, Lanzhou, 730000, Gansu, China. zhangbl@lzu.edu.cn.

ABSTRACT

Background: Protein arginine methyltransferase 5 (PRMT5), a type II PRMT, is highly expressed in some tumors, but its role in hepatocellular carcinoma (HCC) is still unknown.

Methods: PRMT5 level in HCC specimens was determined by immunohistochemical staining and the association with clinicopathologic features was evaluated. PRMT5 was inhibited by AMI-1 (a small molecule inhibitor of PRMTs) or small interference RNA (siRNA). The proliferation of HCC cells was tested by Cell Counting Kit-8, cell migration was evaluated by Transwell assay and cell cycle and apoptosis were analyzed by flow cytometry. The effect of AMI-1 on HCC in vivo was examined by mouse xenograft model.

Results: PRMT5 expression was markedly upregulated in HCC tissues, and correlated inversely with overall patient survival. Knockdown of PRMT5 significantly reduced the proliferation of HCC cells, but did not affect the growth of normal liver cells. Furthermore, β-catenin was identified as a target of PRMT5. Silencing PRMT5 significantly down-regulated the expression of β-catenin and the downstream effector Cyclin D1 in HCC cells. AMI-1 strongly inhibited HCC growth in vivo, increased the ratio of Bax/Bcl-2, and led to apoptosis and loss of migratory activity in several HCC cells. Meanwhile, AMI-1 decreased the expression levels of symmetric dimethylation of H4 (H4R3me2s), a histone mark of PRMT5.

Conclusions: PRMT5 plays an important role in HCC. PRMT5 may be a promising target for HCC therapy.

No MeSH data available.


Related in: MedlinePlus

AMI-1 inhibits HCC cell growth in vitro and in vivo. a The effect of AMI-1 on the proliferation of human HCC cell lines. b The effect of AMI-1 on tumor formation in a nude mouse xenograft model. c The expression of Bax, Bcl-2 and H4R3me2s in HepG2 cells treated with AMI-1 for 72 h. d Densitometric analysis of band intensities. β-actin was loading control. Control refers to vehicle-treated group
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Fig3: AMI-1 inhibits HCC cell growth in vitro and in vivo. a The effect of AMI-1 on the proliferation of human HCC cell lines. b The effect of AMI-1 on tumor formation in a nude mouse xenograft model. c The expression of Bax, Bcl-2 and H4R3me2s in HepG2 cells treated with AMI-1 for 72 h. d Densitometric analysis of band intensities. β-actin was loading control. Control refers to vehicle-treated group

Mentions: AMI-1 has been applied to inhibit type I PRMT (PRMT1, 3, 4, and 6) activity in vitro [26]. Interestingly, we found that AMI-1 also inhibited 84.2 % of type II PRMT5 activity at the tested concentration (nearly 50 µM) [27]. Therefore, we examined the in vitro and in vivo efficacy of AMI-1 on HCC using human HCC cell lines and xenograft mouse models. The concentrations of AMI-1 used for in vitro and in vivo experiments and for enzymatic assay are different, based on our preliminary experiments and previous literatures [27–30]. As shown in Fig. 3a, AMI-1 elicited a significant inhibition on HCC cell growth. In animal tumor models, the tumors were injected with AMI-1 intratumorly (i.t.), because the drug via systemic delivery is easily denatured or degraded. We found that treatment with AMI-1 reduced tumor weight by 65.1 % compared with control-treated animals (Fig. 3b).Fig. 3


Targeting protein arginine methyltransferase 5 inhibits human hepatocellular carcinoma growth via the downregulation of beta-catenin.

Zhang B, Dong S, Li Z, Lu L, Zhang S, Chen X, Cen X, Wu Y - J Transl Med (2015)

AMI-1 inhibits HCC cell growth in vitro and in vivo. a The effect of AMI-1 on the proliferation of human HCC cell lines. b The effect of AMI-1 on tumor formation in a nude mouse xenograft model. c The expression of Bax, Bcl-2 and H4R3me2s in HepG2 cells treated with AMI-1 for 72 h. d Densitometric analysis of band intensities. β-actin was loading control. Control refers to vehicle-treated group
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4635578&req=5

Fig3: AMI-1 inhibits HCC cell growth in vitro and in vivo. a The effect of AMI-1 on the proliferation of human HCC cell lines. b The effect of AMI-1 on tumor formation in a nude mouse xenograft model. c The expression of Bax, Bcl-2 and H4R3me2s in HepG2 cells treated with AMI-1 for 72 h. d Densitometric analysis of band intensities. β-actin was loading control. Control refers to vehicle-treated group
Mentions: AMI-1 has been applied to inhibit type I PRMT (PRMT1, 3, 4, and 6) activity in vitro [26]. Interestingly, we found that AMI-1 also inhibited 84.2 % of type II PRMT5 activity at the tested concentration (nearly 50 µM) [27]. Therefore, we examined the in vitro and in vivo efficacy of AMI-1 on HCC using human HCC cell lines and xenograft mouse models. The concentrations of AMI-1 used for in vitro and in vivo experiments and for enzymatic assay are different, based on our preliminary experiments and previous literatures [27–30]. As shown in Fig. 3a, AMI-1 elicited a significant inhibition on HCC cell growth. In animal tumor models, the tumors were injected with AMI-1 intratumorly (i.t.), because the drug via systemic delivery is easily denatured or degraded. We found that treatment with AMI-1 reduced tumor weight by 65.1 % compared with control-treated animals (Fig. 3b).Fig. 3

Bottom Line: Furthermore, β-catenin was identified as a target of PRMT5.Silencing PRMT5 significantly down-regulated the expression of β-catenin and the downstream effector Cyclin D1 in HCC cells.Meanwhile, AMI-1 decreased the expression levels of symmetric dimethylation of H4 (H4R3me2s), a histone mark of PRMT5.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacology, School of Basic Medical Sciences, Lanzhou University; Key Lab of Preclinical Study for New Drugs of Gansu Province, No 199, Dongang West Road, Lanzhou, 730000, Gansu, China. zhangbl@lzu.edu.cn.

ABSTRACT

Background: Protein arginine methyltransferase 5 (PRMT5), a type II PRMT, is highly expressed in some tumors, but its role in hepatocellular carcinoma (HCC) is still unknown.

Methods: PRMT5 level in HCC specimens was determined by immunohistochemical staining and the association with clinicopathologic features was evaluated. PRMT5 was inhibited by AMI-1 (a small molecule inhibitor of PRMTs) or small interference RNA (siRNA). The proliferation of HCC cells was tested by Cell Counting Kit-8, cell migration was evaluated by Transwell assay and cell cycle and apoptosis were analyzed by flow cytometry. The effect of AMI-1 on HCC in vivo was examined by mouse xenograft model.

Results: PRMT5 expression was markedly upregulated in HCC tissues, and correlated inversely with overall patient survival. Knockdown of PRMT5 significantly reduced the proliferation of HCC cells, but did not affect the growth of normal liver cells. Furthermore, β-catenin was identified as a target of PRMT5. Silencing PRMT5 significantly down-regulated the expression of β-catenin and the downstream effector Cyclin D1 in HCC cells. AMI-1 strongly inhibited HCC growth in vivo, increased the ratio of Bax/Bcl-2, and led to apoptosis and loss of migratory activity in several HCC cells. Meanwhile, AMI-1 decreased the expression levels of symmetric dimethylation of H4 (H4R3me2s), a histone mark of PRMT5.

Conclusions: PRMT5 plays an important role in HCC. PRMT5 may be a promising target for HCC therapy.

No MeSH data available.


Related in: MedlinePlus