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Chemopreventive effects of berberine on intestinal tumor development in Apcmin/+ mice.

Cao H, Song S, Zhang H, Zhang Y, Qu R, Yang B, Jing Y, Hu T, Yan F, Wang B - BMC Gastroenterol (2013)

Bottom Line: The prostaglandin E2 level in the small intestine was detected using ELISA.Furthermore, berberine decreased the activation levels of Wnt and EGFR signaling pathways, and down-regulated COX-2 expression in intestinal tumor cells and prostaglandin E2 production in the small intestine.Down-regulation of Wnt and EGFR signaling pathways and COX-2 expression by berberine may be involved in its anti-tumorigenic effects.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Gastroenterology and Hepatology, General Hospital, Tianjin Medical University, 154 Anshan Road in Heping District, Tianjin 300052, China. gi.tmuh@yeah.net.

ABSTRACT

Background: Berberine, an isoquinoline alkaloid, has shown inhibitory effects on growth of several tumor cell lines in vitro. The aim of this study was to investigate chemopreventive effects of berberine on intestinal tumor development in Apcmin/+ mice.

Methods: Four-week old Apcmin/+ mice were treated with 0.05% or 0.1% berberine in drinking water for twelve weeks. The number and the size of tumors were measured to evaluate intestinal tumor development. Tissue sections were prepared for PCNA and Ki-67 immunostaining to detect cell proliferation, and TUNEL assay and cleaved caspase-3 immunostaining for apoptosis. Western blot analysis and immunostaining were performed to detect the activation of Wnt and epidermal growth factor receptor (EGFR) signaling pathways and COX-2 expression in the intestinal tumor cells. The prostaglandin E2 level in the small intestine was detected using ELISA.

Results: Compared with untreated Apcmin/+ mice, the total numbers of tumors in the small intestine and the colon were reduced by 39.6% and 62.5% in 0.05% and 0.1% berberine-treated mice, respectively. The numbers of tumors in proximal, middle, and distal segments of the small intestine in 0.1% berberine-treated mice were significantly reduced by 53.7%, 55.3%, and 76.5% respectively. Berberine treatment also decreased the numbers of all sizes of tumors (>2 mm, 1-2 mm, and <1 mm) in the small intestine. Berberine suppressed tumor cell proliferation and increased apoptosis. Furthermore, berberine decreased the activation levels of Wnt and EGFR signaling pathways, and down-regulated COX-2 expression in intestinal tumor cells and prostaglandin E2 production in the small intestine.

Conclusions: Berberine inhibits intestinal tumor development, which is correlated with its activity to suppress tumor cell proliferation and increase apoptosis in Apcmin/+ mice. Down-regulation of Wnt and EGFR signaling pathways and COX-2 expression by berberine may be involved in its anti-tumorigenic effects.

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Berberine down-regulates COX-2 expression and PGE2 production. (A-B) Tumors of the distal small intestine from untreated and 0.1% berberine-treated groups were immunostained using COX-2 antibody. Immunoreactivity scoring using a modified semiquantitative scoring system was shown (400×). Scale bars, 50 μm. (C) Protein lysates were prepared from tumors and analyzed by Western blot analysis. Anti-β-actin antibody was used as a protein loading control. The protein band ratio was calculated by comparing the relative density of the protein band on Western blots for COX-2 to that of internal control band from the same mouse. The average ratio in control was set as 100%, the fold change of the ratio in treated mice was shown. (D) PGE2 levels of normal mucosa from untreated and 0.1% berberine-treated groups were detected by ELISA. Columns, means from at least six mice in each group for immunohistochemistry and Western blot analysis, and ten mice in each group for ELISA assay; bars, standard deviation. *, P < 0.01, 0.1% berberine-treated vs untreated Apcmin/+ mice.
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Figure 4: Berberine down-regulates COX-2 expression and PGE2 production. (A-B) Tumors of the distal small intestine from untreated and 0.1% berberine-treated groups were immunostained using COX-2 antibody. Immunoreactivity scoring using a modified semiquantitative scoring system was shown (400×). Scale bars, 50 μm. (C) Protein lysates were prepared from tumors and analyzed by Western blot analysis. Anti-β-actin antibody was used as a protein loading control. The protein band ratio was calculated by comparing the relative density of the protein band on Western blots for COX-2 to that of internal control band from the same mouse. The average ratio in control was set as 100%, the fold change of the ratio in treated mice was shown. (D) PGE2 levels of normal mucosa from untreated and 0.1% berberine-treated groups were detected by ELISA. Columns, means from at least six mice in each group for immunohistochemistry and Western blot analysis, and ten mice in each group for ELISA assay; bars, standard deviation. *, P < 0.01, 0.1% berberine-treated vs untreated Apcmin/+ mice.

Mentions: Over expression of COX-2 and increased PGE2 production were reported to be associated with chronic inflammation and cell proliferation. Berberine significantly down-regulated COX-2 immunoreactivity in small intestinal tumors of Apcmin/+ mice (3.38 ± 0.51 vs 7.60 ± 0.57, P < 0.01, Figure 4A-B). The expression level of COX-2 was significantly decreased in tumors of berberine-treated Apcmin/+ mice, compared with that of untreated group (P < 0.01, Figure 4C). Berberine also decreased PGE2 production, a downstream product of COX-2, in normal mucosa (0.76 ± 0.08 vs 1.06 ± 0.10, P < 0.01; Figure 4D). The results suggest that the inhibition of COX-2 and PGE2 production by berberine may play a role in chemoprevention of intestinal tumorigenesis.


Chemopreventive effects of berberine on intestinal tumor development in Apcmin/+ mice.

Cao H, Song S, Zhang H, Zhang Y, Qu R, Yang B, Jing Y, Hu T, Yan F, Wang B - BMC Gastroenterol (2013)

Berberine down-regulates COX-2 expression and PGE2 production. (A-B) Tumors of the distal small intestine from untreated and 0.1% berberine-treated groups were immunostained using COX-2 antibody. Immunoreactivity scoring using a modified semiquantitative scoring system was shown (400×). Scale bars, 50 μm. (C) Protein lysates were prepared from tumors and analyzed by Western blot analysis. Anti-β-actin antibody was used as a protein loading control. The protein band ratio was calculated by comparing the relative density of the protein band on Western blots for COX-2 to that of internal control band from the same mouse. The average ratio in control was set as 100%, the fold change of the ratio in treated mice was shown. (D) PGE2 levels of normal mucosa from untreated and 0.1% berberine-treated groups were detected by ELISA. Columns, means from at least six mice in each group for immunohistochemistry and Western blot analysis, and ten mice in each group for ELISA assay; bars, standard deviation. *, P < 0.01, 0.1% berberine-treated vs untreated Apcmin/+ mice.
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Related In: Results  -  Collection

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Figure 4: Berberine down-regulates COX-2 expression and PGE2 production. (A-B) Tumors of the distal small intestine from untreated and 0.1% berberine-treated groups were immunostained using COX-2 antibody. Immunoreactivity scoring using a modified semiquantitative scoring system was shown (400×). Scale bars, 50 μm. (C) Protein lysates were prepared from tumors and analyzed by Western blot analysis. Anti-β-actin antibody was used as a protein loading control. The protein band ratio was calculated by comparing the relative density of the protein band on Western blots for COX-2 to that of internal control band from the same mouse. The average ratio in control was set as 100%, the fold change of the ratio in treated mice was shown. (D) PGE2 levels of normal mucosa from untreated and 0.1% berberine-treated groups were detected by ELISA. Columns, means from at least six mice in each group for immunohistochemistry and Western blot analysis, and ten mice in each group for ELISA assay; bars, standard deviation. *, P < 0.01, 0.1% berberine-treated vs untreated Apcmin/+ mice.
Mentions: Over expression of COX-2 and increased PGE2 production were reported to be associated with chronic inflammation and cell proliferation. Berberine significantly down-regulated COX-2 immunoreactivity in small intestinal tumors of Apcmin/+ mice (3.38 ± 0.51 vs 7.60 ± 0.57, P < 0.01, Figure 4A-B). The expression level of COX-2 was significantly decreased in tumors of berberine-treated Apcmin/+ mice, compared with that of untreated group (P < 0.01, Figure 4C). Berberine also decreased PGE2 production, a downstream product of COX-2, in normal mucosa (0.76 ± 0.08 vs 1.06 ± 0.10, P < 0.01; Figure 4D). The results suggest that the inhibition of COX-2 and PGE2 production by berberine may play a role in chemoprevention of intestinal tumorigenesis.

Bottom Line: The prostaglandin E2 level in the small intestine was detected using ELISA.Furthermore, berberine decreased the activation levels of Wnt and EGFR signaling pathways, and down-regulated COX-2 expression in intestinal tumor cells and prostaglandin E2 production in the small intestine.Down-regulation of Wnt and EGFR signaling pathways and COX-2 expression by berberine may be involved in its anti-tumorigenic effects.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Gastroenterology and Hepatology, General Hospital, Tianjin Medical University, 154 Anshan Road in Heping District, Tianjin 300052, China. gi.tmuh@yeah.net.

ABSTRACT

Background: Berberine, an isoquinoline alkaloid, has shown inhibitory effects on growth of several tumor cell lines in vitro. The aim of this study was to investigate chemopreventive effects of berberine on intestinal tumor development in Apcmin/+ mice.

Methods: Four-week old Apcmin/+ mice were treated with 0.05% or 0.1% berberine in drinking water for twelve weeks. The number and the size of tumors were measured to evaluate intestinal tumor development. Tissue sections were prepared for PCNA and Ki-67 immunostaining to detect cell proliferation, and TUNEL assay and cleaved caspase-3 immunostaining for apoptosis. Western blot analysis and immunostaining were performed to detect the activation of Wnt and epidermal growth factor receptor (EGFR) signaling pathways and COX-2 expression in the intestinal tumor cells. The prostaglandin E2 level in the small intestine was detected using ELISA.

Results: Compared with untreated Apcmin/+ mice, the total numbers of tumors in the small intestine and the colon were reduced by 39.6% and 62.5% in 0.05% and 0.1% berberine-treated mice, respectively. The numbers of tumors in proximal, middle, and distal segments of the small intestine in 0.1% berberine-treated mice were significantly reduced by 53.7%, 55.3%, and 76.5% respectively. Berberine treatment also decreased the numbers of all sizes of tumors (>2 mm, 1-2 mm, and <1 mm) in the small intestine. Berberine suppressed tumor cell proliferation and increased apoptosis. Furthermore, berberine decreased the activation levels of Wnt and EGFR signaling pathways, and down-regulated COX-2 expression in intestinal tumor cells and prostaglandin E2 production in the small intestine.

Conclusions: Berberine inhibits intestinal tumor development, which is correlated with its activity to suppress tumor cell proliferation and increase apoptosis in Apcmin/+ mice. Down-regulation of Wnt and EGFR signaling pathways and COX-2 expression by berberine may be involved in its anti-tumorigenic effects.

Show MeSH
Related in: MedlinePlus