Limits...
Anti-tumor effects of Ganoderma lucidum (reishi) in inflammatory breast cancer in in vivo and in vitro models.

Suarez-Arroyo IJ, Rosario-Acevedo R, Aguilar-Perez A, Clemente PL, Cubano LA, Serrano J, Schneider RJ, Martínez-Montemayor MM - PLoS ONE (2013)

Bottom Line: Our previous studies demonstrate these selective anti-cancer effects of Reishi, where IBC cell viability and invasion, as well as the expression of key IBC molecules, including eIF4G is compromised.Thus, herein we define the mechanistic effects of Reishi focusing on the phosphoinositide-3-kinase (PI3K)/AKT/mammalian target of rapamycin (mTOR) pathway, a regulator of cell survival and growth.The present study demonstrates that Reishi treated IBC SUM-149 cells have reduced expression of mTOR downstream effectors at early treatment times, as we observe reduced eIF4G levels coupled with increased levels of eIF4E bound to 4E-BP, with consequential protein synthesis reduction.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemistry, Universidad Central del Caribe, School of Medicine, Bayamón, Puerto Rico, United States of America.

ABSTRACT
The medicinal mushroom Ganoderma lucidum (Reishi) was tested as a potential therapeutic for Inflammatory Breast Cancer (IBC) using in vivo and in vitro IBC models. IBC is a lethal and aggressive form of breast cancer that manifests itself without a typical tumor mass. Studies show that IBC tissue biopsies overexpress E-cadherin and the eukaryotic initiation factor 4GI (eIF4GI), two proteins that are partially responsible for the unique pathological properties of this disease. IBC is treated with a multimodal approach that includes non-targeted systemic chemotherapy, surgery, and radiation. Because of its non-toxic and selective anti-cancer activity, medicinal mushroom extracts have received attention for their use in cancer therapy. Our previous studies demonstrate these selective anti-cancer effects of Reishi, where IBC cell viability and invasion, as well as the expression of key IBC molecules, including eIF4G is compromised. Thus, herein we define the mechanistic effects of Reishi focusing on the phosphoinositide-3-kinase (PI3K)/AKT/mammalian target of rapamycin (mTOR) pathway, a regulator of cell survival and growth. The present study demonstrates that Reishi treated IBC SUM-149 cells have reduced expression of mTOR downstream effectors at early treatment times, as we observe reduced eIF4G levels coupled with increased levels of eIF4E bound to 4E-BP, with consequential protein synthesis reduction. Severe combined immunodeficient mice injected with IBC cells treated with Reishi for 13 weeks show reduced tumor growth and weight by ∼50%, and Reishi treated tumors showed reduced expression of E-cadherin, mTOR, eIF4G, and p70S6K, and activity of extracellular regulated kinase (ERK1/2). Our results provide evidence that Reishi suppresses protein synthesis and tumor growth by affecting survival and proliferative signaling pathways that act on translation, suggesting that Reishi is a potential natural therapeutic for breast and other cancers.

Show MeSH

Related in: MedlinePlus

Reishi reduces tumor growth, tumor weight, and proliferative and mesenchymal marker expression.1.5×106 cells/100µL of SUM-149 cells, were injected into the mammary fat pad of severe combined immunodeficient (SCID) mice. One week following injection, mice were orally gavaged with vehicle, (n = 11) or 28 mg/kg BW Reishi (n = 11) daily for a period of 13 weeks. A. Mice weights were recorded weekly. There were no differences in body weights of mice that received Reishi compared to animals that received vehicle control. B. Tumor volume was recorded weekly using caliper measurements, and measured as described in materials and methods. C. Average tumor volume measurements per week from mice treated with vehicle or Reishi were normalized relative to the average tumor volume measurements from mice treated with vehicle or Reishi obtained at week one. Reishi significantly reduces tumor growth by 58% (P<0.02). D. Tumor weights were obtained at the end of the study. Columns show means ± SEM. Reishi significantly (*P<0.05) reduces tumor weight by 45%. E. Tumors were excised on the 13th week post Reishi, fixed in 10% formalin and embedded in paraffin before immunostaining with antibodies against Ki-67 and vimentin. Reishi treated tumors show reduced size, lower Ki-67 and Vimentin protein expression.
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC3585368&req=5

pone-0057431-g003: Reishi reduces tumor growth, tumor weight, and proliferative and mesenchymal marker expression.1.5×106 cells/100µL of SUM-149 cells, were injected into the mammary fat pad of severe combined immunodeficient (SCID) mice. One week following injection, mice were orally gavaged with vehicle, (n = 11) or 28 mg/kg BW Reishi (n = 11) daily for a period of 13 weeks. A. Mice weights were recorded weekly. There were no differences in body weights of mice that received Reishi compared to animals that received vehicle control. B. Tumor volume was recorded weekly using caliper measurements, and measured as described in materials and methods. C. Average tumor volume measurements per week from mice treated with vehicle or Reishi were normalized relative to the average tumor volume measurements from mice treated with vehicle or Reishi obtained at week one. Reishi significantly reduces tumor growth by 58% (P<0.02). D. Tumor weights were obtained at the end of the study. Columns show means ± SEM. Reishi significantly (*P<0.05) reduces tumor weight by 45%. E. Tumors were excised on the 13th week post Reishi, fixed in 10% formalin and embedded in paraffin before immunostaining with antibodies against Ki-67 and vimentin. Reishi treated tumors show reduced size, lower Ki-67 and Vimentin protein expression.

Mentions: Because Reishi selectively reduces cancer cell viability and invasion, as well as the expression of key proteins involved in the pathogenesis of IBC cells [9] we sought to determine the in vivo efficacy of this extract in SCID mice injected with SUM-149 IBC cells. Mice were injected with IBC cells in Matrigel in their 4th mammary fat pad. When tumors were palpable (∼ one week post-injection), mice were orally gavaged daily with 0 or 28 mg/kg BW Reishi. This concentration is twice the recommended Reishi dose/body weight (1000 mg/daily) for an average adult woman (70 kg). Throughout 13 weeks, the mice were weighed weekly and tumor growth was recorded by precision caliper measurements. There were no differences in body weights (Figure 3A) or food consumption (data not shown) in mice that received Reishi compared to animals that received the vehicle control, which demonstrates that Reishi treatment is not toxic to mice. The drop in body weight at 12 weeks detected in both groups was a result of changing the weighing instrument for that week. However, the mice show similar body weights regardless of this change. In contrast, tumor volume was significantly (>50%) reduced (P<0.02) in the Reishi treated mice compared with mice gavaged daily with vehicle treatment (Figure 3B, 3C). At the end of 13 weeks of daily Reishi treatment, the mice were euthanized and primary tumors and spleens were weighed and collected for subsequent analysis. Reishi treated mice showed a 45% (P<0.05) lower tumor weight values (Figure 3D), while no changes were detected in spleen weights (data not shown). Part of the primary tumor was stored in 10% formalin for tissue paraffin block preparation and subsequent immunohistochemistry, another part was stored in RNAlater™ for real time RT2 profiler PCR array analysis and another part was flash frozen for subsequent western blot analysis of tumor tissue lysates. As depicted in Figure 3E, Reishi treated tumors showed reduced size accompanied by reduced levels of Ki-67 and Vimentin (cell proliferation and mesenchymal markers, respectively) when compared with tumors from mice receiving vehicle treatment. These data are consistent with the results of Figure 1 demonstrating reduced mTOR activity in Reishi treated cancer cells.


Anti-tumor effects of Ganoderma lucidum (reishi) in inflammatory breast cancer in in vivo and in vitro models.

Suarez-Arroyo IJ, Rosario-Acevedo R, Aguilar-Perez A, Clemente PL, Cubano LA, Serrano J, Schneider RJ, Martínez-Montemayor MM - PLoS ONE (2013)

Reishi reduces tumor growth, tumor weight, and proliferative and mesenchymal marker expression.1.5×106 cells/100µL of SUM-149 cells, were injected into the mammary fat pad of severe combined immunodeficient (SCID) mice. One week following injection, mice were orally gavaged with vehicle, (n = 11) or 28 mg/kg BW Reishi (n = 11) daily for a period of 13 weeks. A. Mice weights were recorded weekly. There were no differences in body weights of mice that received Reishi compared to animals that received vehicle control. B. Tumor volume was recorded weekly using caliper measurements, and measured as described in materials and methods. C. Average tumor volume measurements per week from mice treated with vehicle or Reishi were normalized relative to the average tumor volume measurements from mice treated with vehicle or Reishi obtained at week one. Reishi significantly reduces tumor growth by 58% (P<0.02). D. Tumor weights were obtained at the end of the study. Columns show means ± SEM. Reishi significantly (*P<0.05) reduces tumor weight by 45%. E. Tumors were excised on the 13th week post Reishi, fixed in 10% formalin and embedded in paraffin before immunostaining with antibodies against Ki-67 and vimentin. Reishi treated tumors show reduced size, lower Ki-67 and Vimentin protein expression.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3585368&req=5

pone-0057431-g003: Reishi reduces tumor growth, tumor weight, and proliferative and mesenchymal marker expression.1.5×106 cells/100µL of SUM-149 cells, were injected into the mammary fat pad of severe combined immunodeficient (SCID) mice. One week following injection, mice were orally gavaged with vehicle, (n = 11) or 28 mg/kg BW Reishi (n = 11) daily for a period of 13 weeks. A. Mice weights were recorded weekly. There were no differences in body weights of mice that received Reishi compared to animals that received vehicle control. B. Tumor volume was recorded weekly using caliper measurements, and measured as described in materials and methods. C. Average tumor volume measurements per week from mice treated with vehicle or Reishi were normalized relative to the average tumor volume measurements from mice treated with vehicle or Reishi obtained at week one. Reishi significantly reduces tumor growth by 58% (P<0.02). D. Tumor weights were obtained at the end of the study. Columns show means ± SEM. Reishi significantly (*P<0.05) reduces tumor weight by 45%. E. Tumors were excised on the 13th week post Reishi, fixed in 10% formalin and embedded in paraffin before immunostaining with antibodies against Ki-67 and vimentin. Reishi treated tumors show reduced size, lower Ki-67 and Vimentin protein expression.
Mentions: Because Reishi selectively reduces cancer cell viability and invasion, as well as the expression of key proteins involved in the pathogenesis of IBC cells [9] we sought to determine the in vivo efficacy of this extract in SCID mice injected with SUM-149 IBC cells. Mice were injected with IBC cells in Matrigel in their 4th mammary fat pad. When tumors were palpable (∼ one week post-injection), mice were orally gavaged daily with 0 or 28 mg/kg BW Reishi. This concentration is twice the recommended Reishi dose/body weight (1000 mg/daily) for an average adult woman (70 kg). Throughout 13 weeks, the mice were weighed weekly and tumor growth was recorded by precision caliper measurements. There were no differences in body weights (Figure 3A) or food consumption (data not shown) in mice that received Reishi compared to animals that received the vehicle control, which demonstrates that Reishi treatment is not toxic to mice. The drop in body weight at 12 weeks detected in both groups was a result of changing the weighing instrument for that week. However, the mice show similar body weights regardless of this change. In contrast, tumor volume was significantly (>50%) reduced (P<0.02) in the Reishi treated mice compared with mice gavaged daily with vehicle treatment (Figure 3B, 3C). At the end of 13 weeks of daily Reishi treatment, the mice were euthanized and primary tumors and spleens were weighed and collected for subsequent analysis. Reishi treated mice showed a 45% (P<0.05) lower tumor weight values (Figure 3D), while no changes were detected in spleen weights (data not shown). Part of the primary tumor was stored in 10% formalin for tissue paraffin block preparation and subsequent immunohistochemistry, another part was stored in RNAlater™ for real time RT2 profiler PCR array analysis and another part was flash frozen for subsequent western blot analysis of tumor tissue lysates. As depicted in Figure 3E, Reishi treated tumors showed reduced size accompanied by reduced levels of Ki-67 and Vimentin (cell proliferation and mesenchymal markers, respectively) when compared with tumors from mice receiving vehicle treatment. These data are consistent with the results of Figure 1 demonstrating reduced mTOR activity in Reishi treated cancer cells.

Bottom Line: Our previous studies demonstrate these selective anti-cancer effects of Reishi, where IBC cell viability and invasion, as well as the expression of key IBC molecules, including eIF4G is compromised.Thus, herein we define the mechanistic effects of Reishi focusing on the phosphoinositide-3-kinase (PI3K)/AKT/mammalian target of rapamycin (mTOR) pathway, a regulator of cell survival and growth.The present study demonstrates that Reishi treated IBC SUM-149 cells have reduced expression of mTOR downstream effectors at early treatment times, as we observe reduced eIF4G levels coupled with increased levels of eIF4E bound to 4E-BP, with consequential protein synthesis reduction.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemistry, Universidad Central del Caribe, School of Medicine, Bayamón, Puerto Rico, United States of America.

ABSTRACT
The medicinal mushroom Ganoderma lucidum (Reishi) was tested as a potential therapeutic for Inflammatory Breast Cancer (IBC) using in vivo and in vitro IBC models. IBC is a lethal and aggressive form of breast cancer that manifests itself without a typical tumor mass. Studies show that IBC tissue biopsies overexpress E-cadherin and the eukaryotic initiation factor 4GI (eIF4GI), two proteins that are partially responsible for the unique pathological properties of this disease. IBC is treated with a multimodal approach that includes non-targeted systemic chemotherapy, surgery, and radiation. Because of its non-toxic and selective anti-cancer activity, medicinal mushroom extracts have received attention for their use in cancer therapy. Our previous studies demonstrate these selective anti-cancer effects of Reishi, where IBC cell viability and invasion, as well as the expression of key IBC molecules, including eIF4G is compromised. Thus, herein we define the mechanistic effects of Reishi focusing on the phosphoinositide-3-kinase (PI3K)/AKT/mammalian target of rapamycin (mTOR) pathway, a regulator of cell survival and growth. The present study demonstrates that Reishi treated IBC SUM-149 cells have reduced expression of mTOR downstream effectors at early treatment times, as we observe reduced eIF4G levels coupled with increased levels of eIF4E bound to 4E-BP, with consequential protein synthesis reduction. Severe combined immunodeficient mice injected with IBC cells treated with Reishi for 13 weeks show reduced tumor growth and weight by ∼50%, and Reishi treated tumors showed reduced expression of E-cadherin, mTOR, eIF4G, and p70S6K, and activity of extracellular regulated kinase (ERK1/2). Our results provide evidence that Reishi suppresses protein synthesis and tumor growth by affecting survival and proliferative signaling pathways that act on translation, suggesting that Reishi is a potential natural therapeutic for breast and other cancers.

Show MeSH
Related in: MedlinePlus