Limits...
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.

Show MeSH

Related in: MedlinePlus

SENP3 induces the EMT in gastric cancer cells(A) The protein levels of EMT markers and SENP3 in the stable cell lines SGC7901-SENP3 (SENP3 overexpression) and MGC803-sh-SENP3 (SENP3 knockdown). (B) Cell morphology of SGC7901 cells with or without SENP3 overexpression. (C, D) Representative images of wound-healing (C) and transwell assays (D) in SGC7901-MOCK and SGC7901-SENP3 cells were shown. (E, F) Representative images of wound-healing (E) and transwell assays (F) in MGC803-sh-NC and MGC803-sh-SENP3 cells were shown. Transwell assays in SGC7901-SENP3 (8 hours) and MGC803-sh-SENP3 (6 hours). The number of migrated cells was calculated as in Figure 1B,C, and each experiment was repeated 3 times. ***: P < 0.001, **: P < 0.01, *: P < 0.05.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4196186&req=5

Figure 2: SENP3 induces the EMT in gastric cancer cells(A) The protein levels of EMT markers and SENP3 in the stable cell lines SGC7901-SENP3 (SENP3 overexpression) and MGC803-sh-SENP3 (SENP3 knockdown). (B) Cell morphology of SGC7901 cells with or without SENP3 overexpression. (C, D) Representative images of wound-healing (C) and transwell assays (D) in SGC7901-MOCK and SGC7901-SENP3 cells were shown. (E, F) Representative images of wound-healing (E) and transwell assays (F) in MGC803-sh-NC and MGC803-sh-SENP3 cells were shown. Transwell assays in SGC7901-SENP3 (8 hours) and MGC803-sh-SENP3 (6 hours). The number of migrated cells was calculated as in Figure 1B,C, and each experiment was repeated 3 times. ***: P < 0.001, **: P < 0.01, *: P < 0.05.

Mentions: To validate whether SENP3 could induce the EMT in gastric cancer, we established two stable cell lines in which SENP3 was over-expressed in one with low basal SENP3 (SGC7901-SENP3) or knocked-down in one with high basal SENP3 (MGC803-sh-SENP3). The expression of EMT markers was increased in SGC7901-SENP3 cells and decreased in MGC803-sh-SENP3 cells, compared with their respective controls (Fig. 2A). SGC7901-SENP3 cells obtained a mesenchymal spindle-like morphology (Fig. 2B). Scratch wound-healing and transwell assays clearly demonstrated the inductive effect of SENP3 on gastric cancer cell migration, because, compared with their respective controls, gastric cancer cells overexpressing SENP3 migrated markedly faster (Fig. 2C,D), while SENP3 knockdown cells migrated slower (Fig. 2E,F), as confirmed by the quantitative analyses. Viable cell number determination excluded that the difference of migration between the SENP3-interferred and non SENP3-interferred cells was partially due to the differences in cell growth rates (Supplementary Fig. S2).


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 induces the EMT in gastric cancer cells(A) The protein levels of EMT markers and SENP3 in the stable cell lines SGC7901-SENP3 (SENP3 overexpression) and MGC803-sh-SENP3 (SENP3 knockdown). (B) Cell morphology of SGC7901 cells with or without SENP3 overexpression. (C, D) Representative images of wound-healing (C) and transwell assays (D) in SGC7901-MOCK and SGC7901-SENP3 cells were shown. (E, F) Representative images of wound-healing (E) and transwell assays (F) in MGC803-sh-NC and MGC803-sh-SENP3 cells were shown. Transwell assays in SGC7901-SENP3 (8 hours) and MGC803-sh-SENP3 (6 hours). The number of migrated cells was calculated as in Figure 1B,C, and each experiment was repeated 3 times. ***: P < 0.001, **: P < 0.01, *: P < 0.05.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4196186&req=5

Figure 2: SENP3 induces the EMT in gastric cancer cells(A) The protein levels of EMT markers and SENP3 in the stable cell lines SGC7901-SENP3 (SENP3 overexpression) and MGC803-sh-SENP3 (SENP3 knockdown). (B) Cell morphology of SGC7901 cells with or without SENP3 overexpression. (C, D) Representative images of wound-healing (C) and transwell assays (D) in SGC7901-MOCK and SGC7901-SENP3 cells were shown. (E, F) Representative images of wound-healing (E) and transwell assays (F) in MGC803-sh-NC and MGC803-sh-SENP3 cells were shown. Transwell assays in SGC7901-SENP3 (8 hours) and MGC803-sh-SENP3 (6 hours). The number of migrated cells was calculated as in Figure 1B,C, and each experiment was repeated 3 times. ***: P < 0.001, **: P < 0.01, *: P < 0.05.
Mentions: To validate whether SENP3 could induce the EMT in gastric cancer, we established two stable cell lines in which SENP3 was over-expressed in one with low basal SENP3 (SGC7901-SENP3) or knocked-down in one with high basal SENP3 (MGC803-sh-SENP3). The expression of EMT markers was increased in SGC7901-SENP3 cells and decreased in MGC803-sh-SENP3 cells, compared with their respective controls (Fig. 2A). SGC7901-SENP3 cells obtained a mesenchymal spindle-like morphology (Fig. 2B). Scratch wound-healing and transwell assays clearly demonstrated the inductive effect of SENP3 on gastric cancer cell migration, because, compared with their respective controls, gastric cancer cells overexpressing SENP3 migrated markedly faster (Fig. 2C,D), while SENP3 knockdown cells migrated slower (Fig. 2E,F), as confirmed by the quantitative analyses. Viable cell number determination excluded that the difference of migration between the SENP3-interferred and non SENP3-interferred cells was partially due to the differences in cell growth rates (Supplementary Fig. S2).

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