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De-SUMOylation of FOXC2 by SENP3 promotes the epithelial-mesenchymal transition in gastric cancer cells.

Ren YH, Liu KJ, Wang M, Yu YN, Yang K, Chen Q, Yu B, Wang W, Li QW, Wang J, Hou ZY, Fang JY, Yeh ET, Yang J, Yi J - Oncotarget (2014)

Bottom Line: The impact of cellular oxidative stress in promoting the epithelial-mesenchymal transition (EMT) has been noticed.Meanwhile N-cadherin is verified as a target gene of FOXC2, which is transcriptionally activated by a SUMO-less FOXC2.Additionally, reactive oxygen species-induced de-SUMOylation of FOXC2 can be blocked by silencing endogenous SENP3.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemistry and Molecular Cell Biology, Shanghai Key Laboratory of Tumor Microenvironment and Inflammation, Institutes of Medical Sciences, Shanghai Jiao Tong University School of Medicine, Shanghai, China. Institute of Neuroscience, Wenzhou Medical University, School of Medicine, Zhejiang, China. These authors contribute equally to the work.

ABSTRACT
The impact of cellular oxidative stress in promoting the epithelial-mesenchymal transition (EMT) has been noticed. Our previous study shows that SENP3, a redox-sensitive SUMO2/3-specific protease, accumulates in a variety of cancers, but whether SENP3 and SUMOylation involve in the regulation of EMT is unclear. The present study uncovers a novel role of SENP3 in promoting the EMT process in gastric cancer via regulating an EMT-inducing transcription factor, forkhead box C2 (FOXC2). We demonstrate that the expression of mesenchymal marker genes and cell migration ability are enhanced in SENP3-overexpressing gastric cancer cells and attenuated in SENP3-knockdown cells. A nude mouse model and a set of patient's specimens suggest the correlation between SENP3 and gastric cancer metastasis. Biochemical assays identify FOXC2 as a substrate of SENP3. Meanwhile N-cadherin is verified as a target gene of FOXC2, which is transcriptionally activated by a SUMO-less FOXC2. Additionally, reactive oxygen species-induced de-SUMOylation of FOXC2 can be blocked by silencing endogenous SENP3. In conclusion, SENP3, which is increased in gastric cancer cells, potentiates the transcriptional activity of FOXC2 through de-SUMOylation, in favor of the induction of specific mesenchymal gene expression in gastric cancer metastasis.

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SENP3 promotes gastric cancer cell metastasis in vivo(A) The efficiency of SENP3 overexpression in SGC7901-SENP3 cells used for tumorigenesis in nude mice. (B) A mouse (left) and a stomach (right) bearing gastric cancers in the mouse model of orthotopic gastric cancer. (C) The weights of stomach bearing gastric cancer masses in SGC7901-MOCK (n=6) and SGC7901-SENP3 group (n=8).*: P < 0.05. (D) Livers from mice with orthotopically implanted gastric cancers. Red arrows indicated metastatic tumors in SGC7901-SENP3 group. (E) Histology of the hepatic tissue (upper) and the lymph node sections (bottom) derived from the representative mice. Black arrows indicated metastasized tumor cells in hepatic tissue (upper) and the lymph node sections (bottom) and red arrows indicated hepatic cells (upper) and lymph cells (bottom). Scale bar=100 μm. (F) The representative fields of patients' tissues of peri-gastric cancer (Peri-GC, n=21) and gastric cancer with or without lymph node metastasis (GC + LN(+), n=39 and GC + LN(-), n=21) that were examined by SENP3 immunohistochemistry (IHC) (bottom) and hematoxylin /eosin (H&E) histology (upper). Scale bar=100 μm. The percentages of the positive area (middle) and strong positive area (right) of SENP3 immunostaining in gastric cancer tissues were measured by using Zeiss KS400 image analyses software. The average areas of SENP3 immunostaining in epithelial cells were obtained from the whole section of each specimen (mean±SEM). ***: P < 0.001; **: P < 0.01.
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Figure 3: SENP3 promotes gastric cancer cell metastasis in vivo(A) The efficiency of SENP3 overexpression in SGC7901-SENP3 cells used for tumorigenesis in nude mice. (B) A mouse (left) and a stomach (right) bearing gastric cancers in the mouse model of orthotopic gastric cancer. (C) The weights of stomach bearing gastric cancer masses in SGC7901-MOCK (n=6) and SGC7901-SENP3 group (n=8).*: P < 0.05. (D) Livers from mice with orthotopically implanted gastric cancers. Red arrows indicated metastatic tumors in SGC7901-SENP3 group. (E) Histology of the hepatic tissue (upper) and the lymph node sections (bottom) derived from the representative mice. Black arrows indicated metastasized tumor cells in hepatic tissue (upper) and the lymph node sections (bottom) and red arrows indicated hepatic cells (upper) and lymph cells (bottom). Scale bar=100 μm. (F) The representative fields of patients' tissues of peri-gastric cancer (Peri-GC, n=21) and gastric cancer with or without lymph node metastasis (GC + LN(+), n=39 and GC + LN(-), n=21) that were examined by SENP3 immunohistochemistry (IHC) (bottom) and hematoxylin /eosin (H&E) histology (upper). Scale bar=100 μm. The percentages of the positive area (middle) and strong positive area (right) of SENP3 immunostaining in gastric cancer tissues were measured by using Zeiss KS400 image analyses software. The average areas of SENP3 immunostaining in epithelial cells were obtained from the whole section of each specimen (mean±SEM). ***: P < 0.001; **: P < 0.01.

Mentions: An orthotopic gastric cancer metastasis model was established. SGC7901-SENP3 cells (Fig. 3A) were inoculated into nude mice to form tumor xenografts, and the tumors were then recovered, minced into small pieces and implanted beneath the serosa of the stomach of another mouse. Eight weeks later, the tumors grew into large masses in the stomachs (Fig. 3B). The tumors in the SGC7901-SENP3 group were larger than those in the control (Fig. 3C). The livers from each mouse and lymph nodes around the stomach, including pancreaticoduodenal, gastric, caudal mesenteric, jejunal and colic ones, were dissected. The metastasized tumors in the livers were counted under a dissecting microscope. Liver metastases could be found in SGC7901-SENP3 group, but not in the control (Fig. 3D, Table 1). Histological examination demonstrated that the metastasized tumors (Fig. 3E upper, black arrows) grew with the interstitial capsules isolated to the hepatic tissues (red arrows). Lymph nodes metastasis was assessed by histological examination (Fig. 3E, bottom). In many lymph nodes, the metastasized tumors (black arrows) located in the central part, and were relatively isolative of lymphatic tissues (red arrows). The rate of lymph nodes metastasis was markedly higher in the SGC7901-SENP3 group than in the control (Table 2). These data demonstrate that SENP3 promotes gastric cancer cells to metastasize to the liver and surrounding lymph nodes.


De-SUMOylation of FOXC2 by SENP3 promotes the epithelial-mesenchymal transition in gastric cancer cells.

Ren YH, Liu KJ, Wang M, Yu YN, Yang K, Chen Q, Yu B, Wang W, Li QW, Wang J, Hou ZY, Fang JY, Yeh ET, Yang J, Yi J - Oncotarget (2014)

SENP3 promotes gastric cancer cell metastasis in vivo(A) The efficiency of SENP3 overexpression in SGC7901-SENP3 cells used for tumorigenesis in nude mice. (B) A mouse (left) and a stomach (right) bearing gastric cancers in the mouse model of orthotopic gastric cancer. (C) The weights of stomach bearing gastric cancer masses in SGC7901-MOCK (n=6) and SGC7901-SENP3 group (n=8).*: P < 0.05. (D) Livers from mice with orthotopically implanted gastric cancers. Red arrows indicated metastatic tumors in SGC7901-SENP3 group. (E) Histology of the hepatic tissue (upper) and the lymph node sections (bottom) derived from the representative mice. Black arrows indicated metastasized tumor cells in hepatic tissue (upper) and the lymph node sections (bottom) and red arrows indicated hepatic cells (upper) and lymph cells (bottom). Scale bar=100 μm. (F) The representative fields of patients' tissues of peri-gastric cancer (Peri-GC, n=21) and gastric cancer with or without lymph node metastasis (GC + LN(+), n=39 and GC + LN(-), n=21) that were examined by SENP3 immunohistochemistry (IHC) (bottom) and hematoxylin /eosin (H&E) histology (upper). Scale bar=100 μm. The percentages of the positive area (middle) and strong positive area (right) of SENP3 immunostaining in gastric cancer tissues were measured by using Zeiss KS400 image analyses software. The average areas of SENP3 immunostaining in epithelial cells were obtained from the whole section of each specimen (mean±SEM). ***: P < 0.001; **: P < 0.01.
© Copyright Policy - open-access
Related In: Results  -  Collection

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Show All Figures
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Figure 3: SENP3 promotes gastric cancer cell metastasis in vivo(A) The efficiency of SENP3 overexpression in SGC7901-SENP3 cells used for tumorigenesis in nude mice. (B) A mouse (left) and a stomach (right) bearing gastric cancers in the mouse model of orthotopic gastric cancer. (C) The weights of stomach bearing gastric cancer masses in SGC7901-MOCK (n=6) and SGC7901-SENP3 group (n=8).*: P < 0.05. (D) Livers from mice with orthotopically implanted gastric cancers. Red arrows indicated metastatic tumors in SGC7901-SENP3 group. (E) Histology of the hepatic tissue (upper) and the lymph node sections (bottom) derived from the representative mice. Black arrows indicated metastasized tumor cells in hepatic tissue (upper) and the lymph node sections (bottom) and red arrows indicated hepatic cells (upper) and lymph cells (bottom). Scale bar=100 μm. (F) The representative fields of patients' tissues of peri-gastric cancer (Peri-GC, n=21) and gastric cancer with or without lymph node metastasis (GC + LN(+), n=39 and GC + LN(-), n=21) that were examined by SENP3 immunohistochemistry (IHC) (bottom) and hematoxylin /eosin (H&E) histology (upper). Scale bar=100 μm. The percentages of the positive area (middle) and strong positive area (right) of SENP3 immunostaining in gastric cancer tissues were measured by using Zeiss KS400 image analyses software. The average areas of SENP3 immunostaining in epithelial cells were obtained from the whole section of each specimen (mean±SEM). ***: P < 0.001; **: P < 0.01.
Mentions: An orthotopic gastric cancer metastasis model was established. SGC7901-SENP3 cells (Fig. 3A) were inoculated into nude mice to form tumor xenografts, and the tumors were then recovered, minced into small pieces and implanted beneath the serosa of the stomach of another mouse. Eight weeks later, the tumors grew into large masses in the stomachs (Fig. 3B). The tumors in the SGC7901-SENP3 group were larger than those in the control (Fig. 3C). The livers from each mouse and lymph nodes around the stomach, including pancreaticoduodenal, gastric, caudal mesenteric, jejunal and colic ones, were dissected. The metastasized tumors in the livers were counted under a dissecting microscope. Liver metastases could be found in SGC7901-SENP3 group, but not in the control (Fig. 3D, Table 1). Histological examination demonstrated that the metastasized tumors (Fig. 3E upper, black arrows) grew with the interstitial capsules isolated to the hepatic tissues (red arrows). Lymph nodes metastasis was assessed by histological examination (Fig. 3E, bottom). In many lymph nodes, the metastasized tumors (black arrows) located in the central part, and were relatively isolative of lymphatic tissues (red arrows). The rate of lymph nodes metastasis was markedly higher in the SGC7901-SENP3 group than in the control (Table 2). These data demonstrate that SENP3 promotes gastric cancer cells to metastasize to the liver and surrounding lymph nodes.

Bottom Line: The impact of cellular oxidative stress in promoting the epithelial-mesenchymal transition (EMT) has been noticed.Meanwhile N-cadherin is verified as a target gene of FOXC2, which is transcriptionally activated by a SUMO-less FOXC2.Additionally, reactive oxygen species-induced de-SUMOylation of FOXC2 can be blocked by silencing endogenous SENP3.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemistry and Molecular Cell Biology, Shanghai Key Laboratory of Tumor Microenvironment and Inflammation, Institutes of Medical Sciences, Shanghai Jiao Tong University School of Medicine, Shanghai, China. Institute of Neuroscience, Wenzhou Medical University, School of Medicine, Zhejiang, China. These authors contribute equally to the work.

ABSTRACT
The impact of cellular oxidative stress in promoting the epithelial-mesenchymal transition (EMT) has been noticed. Our previous study shows that SENP3, a redox-sensitive SUMO2/3-specific protease, accumulates in a variety of cancers, but whether SENP3 and SUMOylation involve in the regulation of EMT is unclear. The present study uncovers a novel role of SENP3 in promoting the EMT process in gastric cancer via regulating an EMT-inducing transcription factor, forkhead box C2 (FOXC2). We demonstrate that the expression of mesenchymal marker genes and cell migration ability are enhanced in SENP3-overexpressing gastric cancer cells and attenuated in SENP3-knockdown cells. A nude mouse model and a set of patient's specimens suggest the correlation between SENP3 and gastric cancer metastasis. Biochemical assays identify FOXC2 as a substrate of SENP3. Meanwhile N-cadherin is verified as a target gene of FOXC2, which is transcriptionally activated by a SUMO-less FOXC2. Additionally, reactive oxygen species-induced de-SUMOylation of FOXC2 can be blocked by silencing endogenous SENP3. In conclusion, SENP3, which is increased in gastric cancer cells, potentiates the transcriptional activity of FOXC2 through de-SUMOylation, in favor of the induction of specific mesenchymal gene expression in gastric cancer metastasis.

Show MeSH
Related in: MedlinePlus