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Ubiquitin E3 ligase MARCH7 promotes ovarian tumor growth.

Hu J, Meng Y, Yu T, Hu L, Mao M - Oncotarget (2015)

Bottom Line: We found that expression of MARCH7 was higher in ovarian cancer tissues than normal ovarian tissues.Finally, MARCH7 was regulated by miR-101.Thus, MARCH7 is oncogenic and a potential target (oncotarget) for ovarian cancer therapy.

View Article: PubMed Central - PubMed

Affiliation: Department of Obstetrics and Gynecology, Second Affiliated Hospital, Chongqing Medical University, Chongqing, China.

ABSTRACT
Ubiquitin E3 ligase MARCH7 is involved in T cell proliferation and neuronal development. We found that expression of MARCH7 was higher in ovarian cancer tissues than normal ovarian tissues. Silencing MARCH7 decreased cell proliferation, migration, and invasion. Ectopic expression of MARCH7 increased cell proliferation, migration and invasion. Silencing MARCH7 prevented ovarian cancer growth in mice. Silencing MARCH7 inhibited NFkB and Wnt/β-catenin pathway. In agreement, ectopically expressed MARCH7 activated NFkB and Wnt/β-catenin pathways. Finally, MARCH7 was regulated by miR-101. Thus, MARCH7 is oncogenic and a potential target (oncotarget) for ovarian cancer therapy.

No MeSH data available.


Related in: MedlinePlus

(A) The expression of MARCH7 protein level in SKOV3 cells was regulated by TGF-β1, TNF-α, IL-1β, PTDC, NFkB P50 and NFkB P65.(B) The protein expression level in SKOV3 and A2780 cells of NFkB P65, NFkB P50, E-cadherin and β-catenin was detected by western blot. (C-I) A putative binding site targeted by miR-101 was predicted to be located in 3′UTR of MARCH7 mRNA. (C-II)SKOV3 cells were co-transfected with miR-101 mimics or control RNA [negative control (NC)] with luciferase reporter plasmids containing either wild type (pMIR-MARCH7-3UTR) or mutant 3′UTR (pMIR-MARCH7-3UTRm) of MARCH7 gene. Luciferase expression was measured. The fold changes of relative luciferase activity in miR-101 mimics with indicated plasmids transfected cells were normalized to NC with corresponding indicated plasmids transtected cells, respectively. (C-III)The expression of MARCH7 protein levels was detected by western blot. (D) Ovarian cancer SKOV3 cells migration ability was detected by the wound healing assay. (E) Ovarian cancer SKOV3 cells invasion ability was detected by Matrigel invasion assays. (F) Cell proliferation was determined by CCK-8 assay. * p<0.05, and **p<0.001.
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Figure 4: (A) The expression of MARCH7 protein level in SKOV3 cells was regulated by TGF-β1, TNF-α, IL-1β, PTDC, NFkB P50 and NFkB P65.(B) The protein expression level in SKOV3 and A2780 cells of NFkB P65, NFkB P50, E-cadherin and β-catenin was detected by western blot. (C-I) A putative binding site targeted by miR-101 was predicted to be located in 3′UTR of MARCH7 mRNA. (C-II)SKOV3 cells were co-transfected with miR-101 mimics or control RNA [negative control (NC)] with luciferase reporter plasmids containing either wild type (pMIR-MARCH7-3UTR) or mutant 3′UTR (pMIR-MARCH7-3UTRm) of MARCH7 gene. Luciferase expression was measured. The fold changes of relative luciferase activity in miR-101 mimics with indicated plasmids transfected cells were normalized to NC with corresponding indicated plasmids transtected cells, respectively. (C-III)The expression of MARCH7 protein levels was detected by western blot. (D) Ovarian cancer SKOV3 cells migration ability was detected by the wound healing assay. (E) Ovarian cancer SKOV3 cells invasion ability was detected by Matrigel invasion assays. (F) Cell proliferation was determined by CCK-8 assay. * p<0.05, and **p<0.001.

Mentions: Transforming growth factor (TGF)-β1, tumor necrosis factor-alpha (TNF-α) and Interleukin-1β are expressed in ovarian cancer, which can promote ovarian tumorigenesis through an inflammatory response [6-8]. Hence, we explored the possibility of whether TGF-β1, TNF-α, and interleukin-1β can mediate MARCH7 expression in SKOV3 cells. Ovarian cancer SKOV3 cells were treated with TGF-β1 (0, 10, 20, 30 ng/mL), TNF-α (0, 10, 20, 30 ng/mL) or interleukin-1β (0, 10, 20, 30 ng/mL) for 48 hours. TGF-β1 increased the mRNA and protein level of MARCH7 at 10 ng/mL than that untreated. However, the mRNA and protein levels were lower in cells treated with TGF-β1 20 ng/mL or 30 ng/mL. The changes in the mRNA and protein levels of MARCH7 in response to TNF-α or interleukin-1β was similar to that with TGF-β1 treatment at (0, 10, 20, 30 ng/mL) (Fig. 4A and Fig. S1A-C). Our results indicated that TGF-β1, TNF-α, and interleukin-1β regulated MARCH7 expression to promote tumor metastasis of ovarian cancer.


Ubiquitin E3 ligase MARCH7 promotes ovarian tumor growth.

Hu J, Meng Y, Yu T, Hu L, Mao M - Oncotarget (2015)

(A) The expression of MARCH7 protein level in SKOV3 cells was regulated by TGF-β1, TNF-α, IL-1β, PTDC, NFkB P50 and NFkB P65.(B) The protein expression level in SKOV3 and A2780 cells of NFkB P65, NFkB P50, E-cadherin and β-catenin was detected by western blot. (C-I) A putative binding site targeted by miR-101 was predicted to be located in 3′UTR of MARCH7 mRNA. (C-II)SKOV3 cells were co-transfected with miR-101 mimics or control RNA [negative control (NC)] with luciferase reporter plasmids containing either wild type (pMIR-MARCH7-3UTR) or mutant 3′UTR (pMIR-MARCH7-3UTRm) of MARCH7 gene. Luciferase expression was measured. The fold changes of relative luciferase activity in miR-101 mimics with indicated plasmids transfected cells were normalized to NC with corresponding indicated plasmids transtected cells, respectively. (C-III)The expression of MARCH7 protein levels was detected by western blot. (D) Ovarian cancer SKOV3 cells migration ability was detected by the wound healing assay. (E) Ovarian cancer SKOV3 cells invasion ability was detected by Matrigel invasion assays. (F) Cell proliferation was determined by CCK-8 assay. * p<0.05, and **p<0.001.
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Figure 4: (A) The expression of MARCH7 protein level in SKOV3 cells was regulated by TGF-β1, TNF-α, IL-1β, PTDC, NFkB P50 and NFkB P65.(B) The protein expression level in SKOV3 and A2780 cells of NFkB P65, NFkB P50, E-cadherin and β-catenin was detected by western blot. (C-I) A putative binding site targeted by miR-101 was predicted to be located in 3′UTR of MARCH7 mRNA. (C-II)SKOV3 cells were co-transfected with miR-101 mimics or control RNA [negative control (NC)] with luciferase reporter plasmids containing either wild type (pMIR-MARCH7-3UTR) or mutant 3′UTR (pMIR-MARCH7-3UTRm) of MARCH7 gene. Luciferase expression was measured. The fold changes of relative luciferase activity in miR-101 mimics with indicated plasmids transfected cells were normalized to NC with corresponding indicated plasmids transtected cells, respectively. (C-III)The expression of MARCH7 protein levels was detected by western blot. (D) Ovarian cancer SKOV3 cells migration ability was detected by the wound healing assay. (E) Ovarian cancer SKOV3 cells invasion ability was detected by Matrigel invasion assays. (F) Cell proliferation was determined by CCK-8 assay. * p<0.05, and **p<0.001.
Mentions: Transforming growth factor (TGF)-β1, tumor necrosis factor-alpha (TNF-α) and Interleukin-1β are expressed in ovarian cancer, which can promote ovarian tumorigenesis through an inflammatory response [6-8]. Hence, we explored the possibility of whether TGF-β1, TNF-α, and interleukin-1β can mediate MARCH7 expression in SKOV3 cells. Ovarian cancer SKOV3 cells were treated with TGF-β1 (0, 10, 20, 30 ng/mL), TNF-α (0, 10, 20, 30 ng/mL) or interleukin-1β (0, 10, 20, 30 ng/mL) for 48 hours. TGF-β1 increased the mRNA and protein level of MARCH7 at 10 ng/mL than that untreated. However, the mRNA and protein levels were lower in cells treated with TGF-β1 20 ng/mL or 30 ng/mL. The changes in the mRNA and protein levels of MARCH7 in response to TNF-α or interleukin-1β was similar to that with TGF-β1 treatment at (0, 10, 20, 30 ng/mL) (Fig. 4A and Fig. S1A-C). Our results indicated that TGF-β1, TNF-α, and interleukin-1β regulated MARCH7 expression to promote tumor metastasis of ovarian cancer.

Bottom Line: We found that expression of MARCH7 was higher in ovarian cancer tissues than normal ovarian tissues.Finally, MARCH7 was regulated by miR-101.Thus, MARCH7 is oncogenic and a potential target (oncotarget) for ovarian cancer therapy.

View Article: PubMed Central - PubMed

Affiliation: Department of Obstetrics and Gynecology, Second Affiliated Hospital, Chongqing Medical University, Chongqing, China.

ABSTRACT
Ubiquitin E3 ligase MARCH7 is involved in T cell proliferation and neuronal development. We found that expression of MARCH7 was higher in ovarian cancer tissues than normal ovarian tissues. Silencing MARCH7 decreased cell proliferation, migration, and invasion. Ectopic expression of MARCH7 increased cell proliferation, migration and invasion. Silencing MARCH7 prevented ovarian cancer growth in mice. Silencing MARCH7 inhibited NFkB and Wnt/β-catenin pathway. In agreement, ectopically expressed MARCH7 activated NFkB and Wnt/β-catenin pathways. Finally, MARCH7 was regulated by miR-101. Thus, MARCH7 is oncogenic and a potential target (oncotarget) for ovarian cancer therapy.

No MeSH data available.


Related in: MedlinePlus