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OGA heterozygosity suppresses intestinal tumorigenesis in Apc(min/+) mice.

Yang YR, Jang HJ, Yoon S, Lee YH, Nam D, Kim IS, Lee H, Kim H, Choi JH, Kang BH, Ryu SH, Suh PG - Oncogenesis (2014)

Bottom Line: In two independent microarray data sets, the expression of OGA and OGT was significantly associated with poor cancer-specific survival of colorectal cancer (CRC) patients.Apc(min/+) OGA(+/-) mice exhibited a significantly increased survival rate compared with Apc(min/+) mice.However, the knockout of OGA did not affect Wnt/β-catenin signaling.

View Article: PubMed Central - PubMed

Affiliation: School of Life Sciences, Ulsan National Institute of Science and Technology, Ulsan, Republic of Korea.

ABSTRACT
Emerging evidence suggests that aberrant O-GlcNAcylation is associated with tumorigenesis. Many oncogenic factors are O-GlcNAcylated, which modulates their functions. However, it remains unclear how O-GlcNAcylation and O-GlcNAc cycling enzymes, O-GlcNAc transferase (OGT) and O-GlcNAcase (OGA), affect the development of cancer in animal models. In this study, we show that reduced level of OGA attenuates colorectal tumorigenesis induced by Adenomatous polyposis coli (Apc) mutation. The levels of O-GlcNAcylation and O-GlcNAc cycling enzymes were simultaneously upregulated in intestinal adenomas from mice, and in human patients. In two independent microarray data sets, the expression of OGA and OGT was significantly associated with poor cancer-specific survival of colorectal cancer (CRC) patients. In addition, OGA heterozygosity, which results in increased levels of O-GlcNAcylation, attenuated intestinal tumor formation in the Apc(min/+) background. Apc(min/+) OGA(+/-) mice exhibited a significantly increased survival rate compared with Apc(min/+) mice. Consistent with this, Apc(min/+) OGA(+/-) mice expressed lower levels of Wnt target genes than Apc(min/+). However, the knockout of OGA did not affect Wnt/β-catenin signaling. Overall, these findings suggest that OGA is crucial for tumor growth in CRC independently of Wnt/β-catenin signaling.

No MeSH data available.


Related in: MedlinePlus

OGA heterozygosity attenuates APC-mediated intestinal tumorigenesis. (a) The expression of OGA was confirmed in OGA+/− intestines. O-GlcNAc levels were elevated, and OGT was downregulated in OGA+/− intestines. (b) The number (left) and size (right) of intestinal tumors. (c) Gross appearance of the small intestine (upper), and hematoxylin and eosin staining (lower) of intestinal adenoma sections from Apcmin/+ and Apcmin/+OGA+/− mice at 20 weeks. n=8 mice per group. Error bars represent ±s.e.m. **P<0.005, *P<0.05 (Student's t-test). (d) Survival rate of Apcmin/+ and Apcmin/+OGA+/− mice (Kaplan–Meier log-rank, P=0.0001, n=24 and n=15, respectively). Apcmin/+ mice n=24; Apcmin/+OGA+/− mice n=15. (e) The mRNA levels of Wnt target genes were analyzed using RT2 Profiler PCR Arrays (Qiagen, Valencia, CA, USA). Mice were killed at 8, 16 or 20 weeks and the mRNA levels of (f) Axin2 and (g) Jun were analyzed by qPCR. The primer sequences (mouse) used were: Axin2 forward, 5′-ACTCTGGAGGCTTTCGTTTG-3′ Axin2 reverse, 5′-TTAAGTCAGCAGGGGCTCAT-3′ Jun forward, 5′-CCTTCTACGACGATGCCCTC-3′ Jun reverse, 5′-GGTTCAAGGTCATGCTCTGTTT-3′ GAPDH forward, 5′-AGGTCGGTGTGAACGGATTTG-3′ and GAPDH reverse, 5′-TGTAGACCATGTAGTTGAGGTCA-3′. Error bars represent the s.d. (n=3); **P<0.005, *P<0.05 (Student's t-test).
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fig3: OGA heterozygosity attenuates APC-mediated intestinal tumorigenesis. (a) The expression of OGA was confirmed in OGA+/− intestines. O-GlcNAc levels were elevated, and OGT was downregulated in OGA+/− intestines. (b) The number (left) and size (right) of intestinal tumors. (c) Gross appearance of the small intestine (upper), and hematoxylin and eosin staining (lower) of intestinal adenoma sections from Apcmin/+ and Apcmin/+OGA+/− mice at 20 weeks. n=8 mice per group. Error bars represent ±s.e.m. **P<0.005, *P<0.05 (Student's t-test). (d) Survival rate of Apcmin/+ and Apcmin/+OGA+/− mice (Kaplan–Meier log-rank, P=0.0001, n=24 and n=15, respectively). Apcmin/+ mice n=24; Apcmin/+OGA+/− mice n=15. (e) The mRNA levels of Wnt target genes were analyzed using RT2 Profiler PCR Arrays (Qiagen, Valencia, CA, USA). Mice were killed at 8, 16 or 20 weeks and the mRNA levels of (f) Axin2 and (g) Jun were analyzed by qPCR. The primer sequences (mouse) used were: Axin2 forward, 5′-ACTCTGGAGGCTTTCGTTTG-3′ Axin2 reverse, 5′-TTAAGTCAGCAGGGGCTCAT-3′ Jun forward, 5′-CCTTCTACGACGATGCCCTC-3′ Jun reverse, 5′-GGTTCAAGGTCATGCTCTGTTT-3′ GAPDH forward, 5′-AGGTCGGTGTGAACGGATTTG-3′ and GAPDH reverse, 5′-TGTAGACCATGTAGTTGAGGTCA-3′. Error bars represent the s.d. (n=3); **P<0.005, *P<0.05 (Student's t-test).

Mentions: We next used OGA+/− mice to explore the effect of increased OGT and OGA expression on colorectal tumorigenesis. Apcmin/+ mice were crossed with OGA+/− mice. The intestines of OGA+/− mice showed reduced levels of both OGA and OGT expression compared with control mice (Figure 3a). We demonstrated previously that OGA−/− mice displayed perinatal lethality, and that OGA−/− mouse embryonic fibroblasts downregulate OGT, which might compensate for the increased levels of O-GlcNAcylation.15 Therefore, OGA+/− mice are the correct model for assessing the effect of reduced OGT and OGA expression on colorectal tumorigenesis. A quantitative analysis of tumor burden revealed a significant twofold reduction in the number and size of adenomas in the small intestine of Apcmin/+OGA+/− compared with Apcmin/+ mice (Figure 3a). Apcmin/+OGA+/− mice displayed fewer and smaller tumors along the intestinal tract (Figures 3b and c). Apcmin/+OGA+/− mice exhibited a significantly increased survival rate compared with Apcmin/+ mice, and all Apcmin/+ mice died within 8 months. In contrast, Apcmin/+OGA+/− mice showed dramatically increased survival rates (Figure 3d).


OGA heterozygosity suppresses intestinal tumorigenesis in Apc(min/+) mice.

Yang YR, Jang HJ, Yoon S, Lee YH, Nam D, Kim IS, Lee H, Kim H, Choi JH, Kang BH, Ryu SH, Suh PG - Oncogenesis (2014)

OGA heterozygosity attenuates APC-mediated intestinal tumorigenesis. (a) The expression of OGA was confirmed in OGA+/− intestines. O-GlcNAc levels were elevated, and OGT was downregulated in OGA+/− intestines. (b) The number (left) and size (right) of intestinal tumors. (c) Gross appearance of the small intestine (upper), and hematoxylin and eosin staining (lower) of intestinal adenoma sections from Apcmin/+ and Apcmin/+OGA+/− mice at 20 weeks. n=8 mice per group. Error bars represent ±s.e.m. **P<0.005, *P<0.05 (Student's t-test). (d) Survival rate of Apcmin/+ and Apcmin/+OGA+/− mice (Kaplan–Meier log-rank, P=0.0001, n=24 and n=15, respectively). Apcmin/+ mice n=24; Apcmin/+OGA+/− mice n=15. (e) The mRNA levels of Wnt target genes were analyzed using RT2 Profiler PCR Arrays (Qiagen, Valencia, CA, USA). Mice were killed at 8, 16 or 20 weeks and the mRNA levels of (f) Axin2 and (g) Jun were analyzed by qPCR. The primer sequences (mouse) used were: Axin2 forward, 5′-ACTCTGGAGGCTTTCGTTTG-3′ Axin2 reverse, 5′-TTAAGTCAGCAGGGGCTCAT-3′ Jun forward, 5′-CCTTCTACGACGATGCCCTC-3′ Jun reverse, 5′-GGTTCAAGGTCATGCTCTGTTT-3′ GAPDH forward, 5′-AGGTCGGTGTGAACGGATTTG-3′ and GAPDH reverse, 5′-TGTAGACCATGTAGTTGAGGTCA-3′. Error bars represent the s.d. (n=3); **P<0.005, *P<0.05 (Student's t-test).
© Copyright Policy - open-access
Related In: Results  -  Collection

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fig3: OGA heterozygosity attenuates APC-mediated intestinal tumorigenesis. (a) The expression of OGA was confirmed in OGA+/− intestines. O-GlcNAc levels were elevated, and OGT was downregulated in OGA+/− intestines. (b) The number (left) and size (right) of intestinal tumors. (c) Gross appearance of the small intestine (upper), and hematoxylin and eosin staining (lower) of intestinal adenoma sections from Apcmin/+ and Apcmin/+OGA+/− mice at 20 weeks. n=8 mice per group. Error bars represent ±s.e.m. **P<0.005, *P<0.05 (Student's t-test). (d) Survival rate of Apcmin/+ and Apcmin/+OGA+/− mice (Kaplan–Meier log-rank, P=0.0001, n=24 and n=15, respectively). Apcmin/+ mice n=24; Apcmin/+OGA+/− mice n=15. (e) The mRNA levels of Wnt target genes were analyzed using RT2 Profiler PCR Arrays (Qiagen, Valencia, CA, USA). Mice were killed at 8, 16 or 20 weeks and the mRNA levels of (f) Axin2 and (g) Jun were analyzed by qPCR. The primer sequences (mouse) used were: Axin2 forward, 5′-ACTCTGGAGGCTTTCGTTTG-3′ Axin2 reverse, 5′-TTAAGTCAGCAGGGGCTCAT-3′ Jun forward, 5′-CCTTCTACGACGATGCCCTC-3′ Jun reverse, 5′-GGTTCAAGGTCATGCTCTGTTT-3′ GAPDH forward, 5′-AGGTCGGTGTGAACGGATTTG-3′ and GAPDH reverse, 5′-TGTAGACCATGTAGTTGAGGTCA-3′. Error bars represent the s.d. (n=3); **P<0.005, *P<0.05 (Student's t-test).
Mentions: We next used OGA+/− mice to explore the effect of increased OGT and OGA expression on colorectal tumorigenesis. Apcmin/+ mice were crossed with OGA+/− mice. The intestines of OGA+/− mice showed reduced levels of both OGA and OGT expression compared with control mice (Figure 3a). We demonstrated previously that OGA−/− mice displayed perinatal lethality, and that OGA−/− mouse embryonic fibroblasts downregulate OGT, which might compensate for the increased levels of O-GlcNAcylation.15 Therefore, OGA+/− mice are the correct model for assessing the effect of reduced OGT and OGA expression on colorectal tumorigenesis. A quantitative analysis of tumor burden revealed a significant twofold reduction in the number and size of adenomas in the small intestine of Apcmin/+OGA+/− compared with Apcmin/+ mice (Figure 3a). Apcmin/+OGA+/− mice displayed fewer and smaller tumors along the intestinal tract (Figures 3b and c). Apcmin/+OGA+/− mice exhibited a significantly increased survival rate compared with Apcmin/+ mice, and all Apcmin/+ mice died within 8 months. In contrast, Apcmin/+OGA+/− mice showed dramatically increased survival rates (Figure 3d).

Bottom Line: In two independent microarray data sets, the expression of OGA and OGT was significantly associated with poor cancer-specific survival of colorectal cancer (CRC) patients.Apc(min/+) OGA(+/-) mice exhibited a significantly increased survival rate compared with Apc(min/+) mice.However, the knockout of OGA did not affect Wnt/β-catenin signaling.

View Article: PubMed Central - PubMed

Affiliation: School of Life Sciences, Ulsan National Institute of Science and Technology, Ulsan, Republic of Korea.

ABSTRACT
Emerging evidence suggests that aberrant O-GlcNAcylation is associated with tumorigenesis. Many oncogenic factors are O-GlcNAcylated, which modulates their functions. However, it remains unclear how O-GlcNAcylation and O-GlcNAc cycling enzymes, O-GlcNAc transferase (OGT) and O-GlcNAcase (OGA), affect the development of cancer in animal models. In this study, we show that reduced level of OGA attenuates colorectal tumorigenesis induced by Adenomatous polyposis coli (Apc) mutation. The levels of O-GlcNAcylation and O-GlcNAc cycling enzymes were simultaneously upregulated in intestinal adenomas from mice, and in human patients. In two independent microarray data sets, the expression of OGA and OGT was significantly associated with poor cancer-specific survival of colorectal cancer (CRC) patients. In addition, OGA heterozygosity, which results in increased levels of O-GlcNAcylation, attenuated intestinal tumor formation in the Apc(min/+) background. Apc(min/+) OGA(+/-) mice exhibited a significantly increased survival rate compared with Apc(min/+) mice. Consistent with this, Apc(min/+) OGA(+/-) mice expressed lower levels of Wnt target genes than Apc(min/+). However, the knockout of OGA did not affect Wnt/β-catenin signaling. Overall, these findings suggest that OGA is crucial for tumor growth in CRC independently of Wnt/β-catenin signaling.

No MeSH data available.


Related in: MedlinePlus