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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: Knockdown of PRMT5 significantly reduced the proliferation of HCC cells, but did not affect the growth of normal liver cells.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 promotes the apoptosis and decreases migratory activity of HCC cells. a HCC cells were treated with vehicle or AMI-I and then stained by Annexin V-fluorescein isothicyanate (FITC) and propidium iodide (PI), followed by flow cytometry analysis. b AMI-1 decreased migratory activity of HCC cells measured by Transwell assay. Representative photos of stained cells are shown. Original magnification ×200. Control refers to vehicle-treated group
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Fig4: AMI-1 promotes the apoptosis and decreases migratory activity of HCC cells. a HCC cells were treated with vehicle or AMI-I and then stained by Annexin V-fluorescein isothicyanate (FITC) and propidium iodide (PI), followed by flow cytometry analysis. b AMI-1 decreased migratory activity of HCC cells measured by Transwell assay. Representative photos of stained cells are shown. Original magnification ×200. Control refers to vehicle-treated group

Mentions: Inhibition of PRMT5 overexpression can induce apoptosis in different types of cancer [24, 31, 32]. To investigate the effect of AMI-1 on HCC cell viability, Bel-7402 and HepG2 cells were treated with either AMI-1 or vehicle only, and apoptosis was determined by Annexin V-FITC/propidium iodide staining and flow cytometry. As shown in Fig. 4a, PRMT5 inhibition by AMI-1 resulted in the induction of apoptosis/death in both cells compared with control cells. In addition, transwell assay showed that treatment of HCC cells with AMI-1 resulted in marked reduction in migration activity compared with control group (Fig. 4b). Furthermore, we employed siRNA to knockdown PRMT5 in HCC cells and found that PRMT5 siRNA increased the apoptosis while decreased the migration of HCC cells (Additional file 1: Figure 3).Fig. 4


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 promotes the apoptosis and decreases migratory activity of HCC cells. a HCC cells were treated with vehicle or AMI-I and then stained by Annexin V-fluorescein isothicyanate (FITC) and propidium iodide (PI), followed by flow cytometry analysis. b AMI-1 decreased migratory activity of HCC cells measured by Transwell assay. Representative photos of stained cells are shown. Original magnification ×200. 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

Fig4: AMI-1 promotes the apoptosis and decreases migratory activity of HCC cells. a HCC cells were treated with vehicle or AMI-I and then stained by Annexin V-fluorescein isothicyanate (FITC) and propidium iodide (PI), followed by flow cytometry analysis. b AMI-1 decreased migratory activity of HCC cells measured by Transwell assay. Representative photos of stained cells are shown. Original magnification ×200. Control refers to vehicle-treated group
Mentions: Inhibition of PRMT5 overexpression can induce apoptosis in different types of cancer [24, 31, 32]. To investigate the effect of AMI-1 on HCC cell viability, Bel-7402 and HepG2 cells were treated with either AMI-1 or vehicle only, and apoptosis was determined by Annexin V-FITC/propidium iodide staining and flow cytometry. As shown in Fig. 4a, PRMT5 inhibition by AMI-1 resulted in the induction of apoptosis/death in both cells compared with control cells. In addition, transwell assay showed that treatment of HCC cells with AMI-1 resulted in marked reduction in migration activity compared with control group (Fig. 4b). Furthermore, we employed siRNA to knockdown PRMT5 in HCC cells and found that PRMT5 siRNA increased the apoptosis while decreased the migration of HCC cells (Additional file 1: Figure 3).Fig. 4

Bottom Line: Knockdown of PRMT5 significantly reduced the proliferation of HCC cells, but did not affect the growth of normal liver cells.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