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Low molecular weight protein tyrosine phosphatase (LMWPTP) upregulation mediates malignant potential in colorectal cancer.

Hoekstra E, Kodach LL, Das AM, Ruela-de-Sousa RR, Ferreira CV, Hardwick JC, van der Woude CJ, Peppelenbosch MP, Ten Hagen TL, Fuhler GM - Oncotarget (2015)

Bottom Line: In the present study, we show that LMWPTP expression is not only significantly increased in colorectal cancer (CRC), but also follows a step-wise increase in different levels of dysplasia.Chemical inhibition of LMWPTP significantly reduces CRC growth.In conclusion, this study shows that LMWPTP is not only overexpressed in colorectal cancer, but it is correlated with the malignant potential of this cancer, suggesting that this phosphatase may act as a predictive biomaker of CRC stage and represents a rational novel target in the treatment of this disease.

View Article: PubMed Central - PubMed

Affiliation: Department of Gastroenterology and Hepatology, Erasmus MC, University Medical Center Rotterdam, 's Gravendijkwa, Rotterdam, The Netherlands.

ABSTRACT
Phosphatases have long been regarded as tumor suppressors, however there is emerging evidence for a tumor initiating role for some phosphatases in several forms of cancer. Low Molecular Weight Protein Tyrosine Phosphatase (LMWPTP; acid phosphatase 1 [ACP1]) is an 18 kDa enzyme that influences the phosphorylation of signaling pathway mediators involved in cancer and is thus postulated to be a tumor-promoting enzyme, but neither unequivocal clinical evidence nor convincing mechanistic actions for a role of LMWPTP have been identified. In the present study, we show that LMWPTP expression is not only significantly increased in colorectal cancer (CRC), but also follows a step-wise increase in different levels of dysplasia. Chemical inhibition of LMWPTP significantly reduces CRC growth. Furthermore, downregulation of LMWPTP in CRC leads to a reduced migration ability in both 2D- and 3D-migration assays, and sensitizes tumor cells to the chemotherapeutic agent 5-FU. In conclusion, this study shows that LMWPTP is not only overexpressed in colorectal cancer, but it is correlated with the malignant potential of this cancer, suggesting that this phosphatase may act as a predictive biomaker of CRC stage and represents a rational novel target in the treatment of this disease.

No MeSH data available.


Related in: MedlinePlus

Modulation of LMWPTP results in impaired migration and invasion in colorectal cancer cells(A, B) HCT116 cell migration was measured by scratch assays, where simple scratch wounds were made using a pipet tip, and pictures are taken at 0 h, 24 h, and 48 h. Persistent area of clear plastic was measured and statistical analysis was performed using student's T-test. (C, D) Two-dimensional migration was analyzed using a ring-barrier system. HCT116 cell migration on gelatin was tracked during 24 h, with locations being captured using time-lapse microscopy every 12 min (x=start, line=cell track) (C). Quantification of migrated path indicates that the total migration and velocity were significantly reduced in LMWPTP knockdown cells. Effective migration and thereby efficiency are even further reduced. (D; *P < 0.05; **P < 0.01; ***P < 0.001). (E, F) Beads were coated with either CACO-2 LMWPTP knockdown or control cells for 24 hours, and embedded in a collagen gel matrix. Cells were allowed to invade the collagen matrix, and pictures were taking at 0 h, 24 h, and 48 h (examples in E). The cell dispersion from the bead (arrow) into the collagen matrix was measured, and a trend towards reduced invasion was observed in LMWPTP knockdown cells. Data represents at least four beads (F).
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Figure 5: Modulation of LMWPTP results in impaired migration and invasion in colorectal cancer cells(A, B) HCT116 cell migration was measured by scratch assays, where simple scratch wounds were made using a pipet tip, and pictures are taken at 0 h, 24 h, and 48 h. Persistent area of clear plastic was measured and statistical analysis was performed using student's T-test. (C, D) Two-dimensional migration was analyzed using a ring-barrier system. HCT116 cell migration on gelatin was tracked during 24 h, with locations being captured using time-lapse microscopy every 12 min (x=start, line=cell track) (C). Quantification of migrated path indicates that the total migration and velocity were significantly reduced in LMWPTP knockdown cells. Effective migration and thereby efficiency are even further reduced. (D; *P < 0.05; **P < 0.01; ***P < 0.001). (E, F) Beads were coated with either CACO-2 LMWPTP knockdown or control cells for 24 hours, and embedded in a collagen gel matrix. Cells were allowed to invade the collagen matrix, and pictures were taking at 0 h, 24 h, and 48 h (examples in E). The cell dispersion from the bead (arrow) into the collagen matrix was measured, and a trend towards reduced invasion was observed in LMWPTP knockdown cells. Data represents at least four beads (F).

Mentions: Colorectal cancer is a frequently fatal disease because of its high propensity to migrate and invade other tissues, preventing curative surgical treatment. Cellular migration is dependent on the tight regulation of assembly and disassembly of focal adhesion sites. This process is mediated the by the formation of a FAK–Src complex, and phosphorylation of FAK-associated substrates such as paxillin and p130cas, all known to be required for cell motility [17]. Our biochemical analysis of LMWPTP deficient cells revealed reduced FAK Tyr-397 phosphorylation in these cells, suggesting that LMWPTP may function in this pathway to promote FAK Tyr-397 phosphorylation and the formation of membrane extensions characteristic of migrating cells (Figure 2A). We therefore investigated the effects of LMWPTP downregulation in colorectal cancer cell lines on their ability to migrate. Confluent plates of CACO-2 and HCT116 cells were scratched using a pipet tip, and cell migration into the wound was assessed after 24 h and 48 h. LMWPTP knock down cells showed a significant delay in the ability to migrate into the empty space (Figure 5A–5B and Supplementary Figures S4A–S4B, N.B. that HCT116 is a slower migrating cell line). To verify the positive role of LMWPTP in cell migration, we used a second, different approach to investigate cellular movement, which does not rely on wounding the CRC monolayer. Using time-lapse microscopy of cell migration we again observed that CACO-2 and HCT116 LMWPTP knockdown cells are significantly impaired in their total migration, and thereby also the cell velocity. Strikingly, the effective migration, which is defined as the directional movement of the cells to the cell-free center, was even more reduced (Figure 5C–5D and Supplementary Figures S4C–S4D), Supplementary Movies). We subsequently went on to assess the role of LMWPTP on migration in a 3D-setting, representing the invasive capacity of these cells. Beads were coated with CACO-2 and HCT116 knockdown or control cells, and were settled in a collagen matrix. Cell dispersion from the bead into the surrounding collagen matrix was measured. Although not reaching statistical significance, we observed a trend towards reduced invasive capacity upon LMWPTP knock down for both cell lines (Figure 5E–5F and Supplementary Figure S4E–S4F). Together these data demonstrate that knocking down LMWPTP in colorectal cancer cells reduces their migratory capacity, and is especially important for directional cell migration.


Low molecular weight protein tyrosine phosphatase (LMWPTP) upregulation mediates malignant potential in colorectal cancer.

Hoekstra E, Kodach LL, Das AM, Ruela-de-Sousa RR, Ferreira CV, Hardwick JC, van der Woude CJ, Peppelenbosch MP, Ten Hagen TL, Fuhler GM - Oncotarget (2015)

Modulation of LMWPTP results in impaired migration and invasion in colorectal cancer cells(A, B) HCT116 cell migration was measured by scratch assays, where simple scratch wounds were made using a pipet tip, and pictures are taken at 0 h, 24 h, and 48 h. Persistent area of clear plastic was measured and statistical analysis was performed using student's T-test. (C, D) Two-dimensional migration was analyzed using a ring-barrier system. HCT116 cell migration on gelatin was tracked during 24 h, with locations being captured using time-lapse microscopy every 12 min (x=start, line=cell track) (C). Quantification of migrated path indicates that the total migration and velocity were significantly reduced in LMWPTP knockdown cells. Effective migration and thereby efficiency are even further reduced. (D; *P < 0.05; **P < 0.01; ***P < 0.001). (E, F) Beads were coated with either CACO-2 LMWPTP knockdown or control cells for 24 hours, and embedded in a collagen gel matrix. Cells were allowed to invade the collagen matrix, and pictures were taking at 0 h, 24 h, and 48 h (examples in E). The cell dispersion from the bead (arrow) into the collagen matrix was measured, and a trend towards reduced invasion was observed in LMWPTP knockdown cells. Data represents at least four beads (F).
© Copyright Policy - open-access
Related In: Results  -  Collection

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Show All Figures
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Figure 5: Modulation of LMWPTP results in impaired migration and invasion in colorectal cancer cells(A, B) HCT116 cell migration was measured by scratch assays, where simple scratch wounds were made using a pipet tip, and pictures are taken at 0 h, 24 h, and 48 h. Persistent area of clear plastic was measured and statistical analysis was performed using student's T-test. (C, D) Two-dimensional migration was analyzed using a ring-barrier system. HCT116 cell migration on gelatin was tracked during 24 h, with locations being captured using time-lapse microscopy every 12 min (x=start, line=cell track) (C). Quantification of migrated path indicates that the total migration and velocity were significantly reduced in LMWPTP knockdown cells. Effective migration and thereby efficiency are even further reduced. (D; *P < 0.05; **P < 0.01; ***P < 0.001). (E, F) Beads were coated with either CACO-2 LMWPTP knockdown or control cells for 24 hours, and embedded in a collagen gel matrix. Cells were allowed to invade the collagen matrix, and pictures were taking at 0 h, 24 h, and 48 h (examples in E). The cell dispersion from the bead (arrow) into the collagen matrix was measured, and a trend towards reduced invasion was observed in LMWPTP knockdown cells. Data represents at least four beads (F).
Mentions: Colorectal cancer is a frequently fatal disease because of its high propensity to migrate and invade other tissues, preventing curative surgical treatment. Cellular migration is dependent on the tight regulation of assembly and disassembly of focal adhesion sites. This process is mediated the by the formation of a FAK–Src complex, and phosphorylation of FAK-associated substrates such as paxillin and p130cas, all known to be required for cell motility [17]. Our biochemical analysis of LMWPTP deficient cells revealed reduced FAK Tyr-397 phosphorylation in these cells, suggesting that LMWPTP may function in this pathway to promote FAK Tyr-397 phosphorylation and the formation of membrane extensions characteristic of migrating cells (Figure 2A). We therefore investigated the effects of LMWPTP downregulation in colorectal cancer cell lines on their ability to migrate. Confluent plates of CACO-2 and HCT116 cells were scratched using a pipet tip, and cell migration into the wound was assessed after 24 h and 48 h. LMWPTP knock down cells showed a significant delay in the ability to migrate into the empty space (Figure 5A–5B and Supplementary Figures S4A–S4B, N.B. that HCT116 is a slower migrating cell line). To verify the positive role of LMWPTP in cell migration, we used a second, different approach to investigate cellular movement, which does not rely on wounding the CRC monolayer. Using time-lapse microscopy of cell migration we again observed that CACO-2 and HCT116 LMWPTP knockdown cells are significantly impaired in their total migration, and thereby also the cell velocity. Strikingly, the effective migration, which is defined as the directional movement of the cells to the cell-free center, was even more reduced (Figure 5C–5D and Supplementary Figures S4C–S4D), Supplementary Movies). We subsequently went on to assess the role of LMWPTP on migration in a 3D-setting, representing the invasive capacity of these cells. Beads were coated with CACO-2 and HCT116 knockdown or control cells, and were settled in a collagen matrix. Cell dispersion from the bead into the surrounding collagen matrix was measured. Although not reaching statistical significance, we observed a trend towards reduced invasive capacity upon LMWPTP knock down for both cell lines (Figure 5E–5F and Supplementary Figure S4E–S4F). Together these data demonstrate that knocking down LMWPTP in colorectal cancer cells reduces their migratory capacity, and is especially important for directional cell migration.

Bottom Line: In the present study, we show that LMWPTP expression is not only significantly increased in colorectal cancer (CRC), but also follows a step-wise increase in different levels of dysplasia.Chemical inhibition of LMWPTP significantly reduces CRC growth.In conclusion, this study shows that LMWPTP is not only overexpressed in colorectal cancer, but it is correlated with the malignant potential of this cancer, suggesting that this phosphatase may act as a predictive biomaker of CRC stage and represents a rational novel target in the treatment of this disease.

View Article: PubMed Central - PubMed

Affiliation: Department of Gastroenterology and Hepatology, Erasmus MC, University Medical Center Rotterdam, 's Gravendijkwa, Rotterdam, The Netherlands.

ABSTRACT
Phosphatases have long been regarded as tumor suppressors, however there is emerging evidence for a tumor initiating role for some phosphatases in several forms of cancer. Low Molecular Weight Protein Tyrosine Phosphatase (LMWPTP; acid phosphatase 1 [ACP1]) is an 18 kDa enzyme that influences the phosphorylation of signaling pathway mediators involved in cancer and is thus postulated to be a tumor-promoting enzyme, but neither unequivocal clinical evidence nor convincing mechanistic actions for a role of LMWPTP have been identified. In the present study, we show that LMWPTP expression is not only significantly increased in colorectal cancer (CRC), but also follows a step-wise increase in different levels of dysplasia. Chemical inhibition of LMWPTP significantly reduces CRC growth. Furthermore, downregulation of LMWPTP in CRC leads to a reduced migration ability in both 2D- and 3D-migration assays, and sensitizes tumor cells to the chemotherapeutic agent 5-FU. In conclusion, this study shows that LMWPTP is not only overexpressed in colorectal cancer, but it is correlated with the malignant potential of this cancer, suggesting that this phosphatase may act as a predictive biomaker of CRC stage and represents a rational novel target in the treatment of this disease.

No MeSH data available.


Related in: MedlinePlus