A. Immunoblot analysis of tuberin and OGG1 protein expression in normal kidney of wild type rats,... more A. Immunoblot analysis of tuberin and OGG1 protein expression in normal kidney of wild type rats, normal and tumor kidney tissue from Eker rats. GAPDH was used as loading control. B. Histograms represent means ± SE (n = 3). Significant difference from wild type rat is indicated by ** < 0.01.<b>Copyright information:</b>Taken from "Tuberin haploinsufficiency is associated with the loss of OGG1 in rat kidney tumors"http://www.molecular-cancer.com/content/7/1/10Molecular Cancer 2008;7():10-10.Published online 24 Jan 2008PMCID:PMC2265742.
Innate immunity is critical for immediate recognition and elimination of invading pathogens or de... more Innate immunity is critical for immediate recognition and elimination of invading pathogens or defense against cancer cell growth. Dysregulation of innate immune systems is associated with the pathogenesis of different types of inflammatory diseases, including cancer. In addition, the maintenance of innate immune cells’ genomic integrity is crucial for the survival of all organisms. Oxidative stress generated from innate immune cells may cause self-inflicted DNA base lesions as well as DNA damage on others neighboring cells, including cancer cells. Oxidative DNA base damage is predominantly repaired by base excision repair (BER). BER process different types of DNA base lesions that are presented in cancer and innate immune cells to maintain genomic integrity. However, mutations in BER genes lead to impaired DNA repair function and cause insufficient genomic integrity. Moreover, several studies have implicated that accumulation of DNA damage leads to chromosomal instability that like...
Annals of transplantation : quarterly of the Polish Transplantation Society, Jan 10, 2015
Background The aim of this study was to evaluate the long-term usefulness of intraportal injectio... more Background The aim of this study was to evaluate the long-term usefulness of intraportal injection of the bone marrow-derived mesenchymal stem cells (BM-MSCs) in limitation of experimentally induced ischemia-reperfusion injury (IRI) in a rat model. Material and Methods Twenty Wistar rats were divided into 3 groups: donor group (n=5), study group (n=10), and control group (n=5). IRI was performed using a modified hanging-weight system after left portal triad occlusion in study group animals. Isolated autologous BM-MSCs were labeled with fluorochrome PKH-26 then intraportally injected into the rats in the study group. Control group animals were intraportally injected with 1 ml of PBS. Follow-up was 3 months, after which animals were sacrificed for histopathological examination. Migration of BM-MSCs into different organs was examined. Results H&E staining of liver tissue sections from "time zero" biopsies did not show many irregularities in structural or histological construc...
In general, detection of peritoneal carcinomatosis (PC) occurs at the late stage when there is no... more In general, detection of peritoneal carcinomatosis (PC) occurs at the late stage when there is no treatment option. In the present study, we designed novel drug delivery systems that are functionalized with anti-CD133 antibodies. The C1, C2 and C3 complexes with cisplatin were introduced into nanotubes, either physically or chemically. The complexes were reacted with anti-CD133 antibody to form the labeled product of A0-o-CX-chem-CD133. Cytotoxicity screening of all the complexes was performed on CHO cells. Data showed that both C2 and C3 Pt-complexes are more cytotoxic than C1. Flow-cytometry analysis showed that nanotubes conjugated to CD133 antibody have the ability to target cells expressing the CD133 antigen which is responsible for the emergence of resistance to chemotherapy and disease recurrence. The shortest survival rate was observed in the control mice group (K3) where no hyperthermic intraperitoneal chemotherapy procedures were used. On the other hand, the longest median...
A. Immunoblot analysis of tuberin and OGG1 in different kidney regions. Different kidney regions ... more A. Immunoblot analysis of tuberin and OGG1 in different kidney regions. Different kidney regions were homogenized and protein extracts were loaded onto 7% SDS-polyacrylamide gels and transfered to PVDF membrane. The membrane was incubated with anti-tuberin or anti-OGG1 followed by different specific HRP-conjugated secondary antibodies. The proteins were visualized by ECL. GAPDH was used as loading control. B. Histograms represent means ± SE (n = 3). Significant difference from wild type rat is indicated by * P < 0.05 and ** P < 0.01.<b>Copyright information:</b>Taken from "Tuberin haploinsufficiency is associated with the loss of OGG1 in rat kidney tumors"http://www.molecular-cancer.com/content/7/1/10Molecular Cancer 2008;7():10-10.Published online 24 Jan 2008PMCID:PMC2265742.
28 Endothelial nitric oxide synthase (eNOS) is the predominant isoform that generates nitric oxid... more 28 Endothelial nitric oxide synthase (eNOS) is the predominant isoform that generates nitric oxide 29 (NO) in the blood vessels. Many different regulators govern eNOS function, including heat 30 shock protein 90 (Hsp90). Hsp90-dependent phosphorylation of eNOS is a critical event that 31 determines eNOS activity. In our earlier study, we demonstrated the occurrence of an Hsp9032 inhibitor kappa B kinase beta (IKKβ) interaction under high glucose environment. In the present 33 study, we have further defined the putative binding domain of IKKβ on Hsp90. Interestingly, 34 IKK binds to the middle domain of Hsp90, which has been shown to interact with eNOS to 35 stimulate its activity. This new finding suggests that there is a tighter regulation of eNOS activity 36 than previously assumed. Furthermore, addition of purified recombinant IKKβ to the eNOS37 Hsp90 complex reduces the eNOS-Hsp90 interaction and eNOS activity, indicating the existence 38 of a competition for Hsp90 between eNOS...
The role of tumor protein 63 (TP63) in regulating insulin receptor substrate 1 (IRS-1) and other ... more The role of tumor protein 63 (TP63) in regulating insulin receptor substrate 1 (IRS-1) and other downstream signal proteins in diabetes has not been characterized. RNAs extracted from kidneys of diabetic mice (db/db) were sequenced to identify genes that are involved in kidney complications. RNA sequence analysis showed more than 4- to 6-fold increases in TP63 expression in the diabetic mice’s kidneys, compared to wild-type mice at age 10 and 12 months old. In addition, the kidneys from diabetic mice showed significant increases in TP63 mRNA and protein expression compared to WT mice. Mouse proximal tubular cells exposed to high glucose (HG) for 48 h showed significant decreases in IRS-1 expression and increases in TP63, compared to cells grown in normal glucose (NG). When TP63 was downregulated by siRNA, significant increases in IRS-1 and activation of AMP-activated protein kinase (AMPK (p-AMPK-Th172)) occurred under NG and HG conditions. Moreover, activation of AMPK by pretreating...
Proteasome inhibitor (PI) therapy has improved the survival of multiple myeloma (MM) patients. Ho... more Proteasome inhibitor (PI) therapy has improved the survival of multiple myeloma (MM) patients. However, inevitably, primary or acquired resistance to PIs leads to disease progression; resistance mechanisms are unclear. Obesity is a risk factor for MM mortality. Oxidized LDL (OxLDL), a central mediator of atherosclerosis that is elevated in metabolic syndrome (co‐occurrence of obesity, insulin resistance, dyslipidemia and hypertension), has been linked to an increased risk of solid cancers and shown to stimulate pro‐oncogenic/survival signaling. We hypothesized that OxLDL is a mediator of chemoresistance and evaluated its effects on MM cell killing by PIs. OxLDL potently suppressed the ability of the boronic acid‐based PIs bortezomib (BTZ) and ixazomib, but not the epoxyketone‐based PI carfilzomib, to kill human MM cell lines and primary cells. OxLDL suppressed BTZ‐induced inhibition of proteasome activity and induction of pro‐apoptotic signaling. These cytoprotective effects were ab...
The role of tumor protein 63 (TP63) in regulating insulin receptor substrate 1 (IRS-1) and other ... more The role of tumor protein 63 (TP63) in regulating insulin receptor substrate 1 (IRS-1) and other downstream signal proteins in diabetes has not been characterized. RNAs extracted from kidneys of diabetic mice (db/db) were sequenced to identify genes that are involved in kidney complications. RNA sequence analysis showed more than 4- to 6-fold increases in TP63 expression in the diabetic mice’s kidneys, compared to wild-type mice at age 10 and 12 months old. In addition, the kidneys from diabetic mice showed significant increases in TP63 mRNA and protein expression compared to WT mice. Mouse proximal tubular cells exposed to high glucose (HG) for 48 h showed significant decreases in IRS-1 expression and increases in TP63, compared to cells grown in normal glucose (NG). When TP63 was downregulated by siRNA, significant increases in IRS-1 and activation of AMP-activated protein kinase (AMPK (p-AMPK-Th172)) occurred under NG and HG conditions. Moreover, activation of AMPK by pretreating...
Epithelial–mesenchymal transition (EMT) plays an important role in tissue fibrosis following chro... more Epithelial–mesenchymal transition (EMT) plays an important role in tissue fibrosis following chronic exposure to hyperglycemia. This study investigates the role of chronic diabetes in regulating tuberin/snail/AMPK to enhance EMT and increase renal fibrosis. A new mouse model of db/db/TSC2+/− was generated by backcrossing db/db mice and TSC2+/− mice. Wild type (WT), db/db, TSC2+/− and dbdb/TSC2+/− mice were sacrificed at ages 6 and 8 months old. Tuberin protein level was significantly decreased in kidneys from diabetic compared to WT mice at both ages. In addition, tuberin and E‐cadherin protein levels were significantly decreased in dbdb/TSC2+/− compared to TSC2+/− and db/db mice. In contrast, p‐PS6K, NFkB, snail, vimentin, fibronectin, and α‐SMA protein levels were significantly increased in dbdb/TSC2+/− compared to db/db and TSC2+/− mice at ages 6 and 8 months. Both downregulation of AMPK by DN‐AMPK and downregulation of tuberin by siRNA resulted in increased NFkB, snail, and fibronectin protein expression and decreased E‐cadherin protein expression in mouse primary renal proximal tubular cells. Interestingly, downregulation of snail by siRNA increased tuberin expression via feedback through activation of AMPK and reversed the expression of epithelial proteins such as E‐cadherin as well as mesenchymal proteins such as fibronectin, NF‐KB, vimentin, and α‐SMA in mouse primary renal proximal tubular cells isolated from kidneys of four mice genotypes. The data show that chronic diabetes significantly decreases tuberin expression and that provides strong evidence that tuberin is a major key protein involved in regulating EMT. These data also demonstrated a novel role for snail in regulating of AMPK/tuberin to enhance EMT and renal cell fibrosis in diabetes.
Insulin resistance is a major risk for development of type 2 diabetes. Several genes involved in ... more Insulin resistance is a major risk for development of type 2 diabetes. Several genes involved in regulation of insulin receptor substrate 1 (IRS1) and lead to chronic diabetes. Our RNA sequence data from kidney of diabetic mice show a new candidate gene involved in the regulation of IRS1. New candidate gene, Tripartite motif protein 29 (TRIM29), acts as an E3 ligase targeting insulin receptor (IR) and insulin receptor substrate 1 (IRS1) for ubiquitin-dependent degradation, resulting in insulin resistance. RNA seq data show that TRIM29 is higher 2.5 and 3.6 fold in kidney cortex of diabetic mice (db/db) at age of 10 and 12 months compared to wild type mice, respectively. Mouse renal proximal tubular epithelial cells treated with high glucose (25mM) for 48 hours showed significant increased in TRIM29 and decreased in IRS-1 compared to cells grown in normal glucose (5mM). Pre-transfected cells with siRNA against TRIM29 resulted in significant increased IRS-1 and cell proliferative mark...
A. Immunoblot analysis of tuberin and OGG1 protein expression in normal kidney of wild type rats,... more A. Immunoblot analysis of tuberin and OGG1 protein expression in normal kidney of wild type rats, normal and tumor kidney tissue from Eker rats. GAPDH was used as loading control. B. Histograms represent means ± SE (n = 3). Significant difference from wild type rat is indicated by ** < 0.01.<b>Copyright information:</b>Taken from "Tuberin haploinsufficiency is associated with the loss of OGG1 in rat kidney tumors"http://www.molecular-cancer.com/content/7/1/10Molecular Cancer 2008;7():10-10.Published online 24 Jan 2008PMCID:PMC2265742.
Innate immunity is critical for immediate recognition and elimination of invading pathogens or de... more Innate immunity is critical for immediate recognition and elimination of invading pathogens or defense against cancer cell growth. Dysregulation of innate immune systems is associated with the pathogenesis of different types of inflammatory diseases, including cancer. In addition, the maintenance of innate immune cells’ genomic integrity is crucial for the survival of all organisms. Oxidative stress generated from innate immune cells may cause self-inflicted DNA base lesions as well as DNA damage on others neighboring cells, including cancer cells. Oxidative DNA base damage is predominantly repaired by base excision repair (BER). BER process different types of DNA base lesions that are presented in cancer and innate immune cells to maintain genomic integrity. However, mutations in BER genes lead to impaired DNA repair function and cause insufficient genomic integrity. Moreover, several studies have implicated that accumulation of DNA damage leads to chromosomal instability that like...
Annals of transplantation : quarterly of the Polish Transplantation Society, Jan 10, 2015
Background The aim of this study was to evaluate the long-term usefulness of intraportal injectio... more Background The aim of this study was to evaluate the long-term usefulness of intraportal injection of the bone marrow-derived mesenchymal stem cells (BM-MSCs) in limitation of experimentally induced ischemia-reperfusion injury (IRI) in a rat model. Material and Methods Twenty Wistar rats were divided into 3 groups: donor group (n=5), study group (n=10), and control group (n=5). IRI was performed using a modified hanging-weight system after left portal triad occlusion in study group animals. Isolated autologous BM-MSCs were labeled with fluorochrome PKH-26 then intraportally injected into the rats in the study group. Control group animals were intraportally injected with 1 ml of PBS. Follow-up was 3 months, after which animals were sacrificed for histopathological examination. Migration of BM-MSCs into different organs was examined. Results H&E staining of liver tissue sections from "time zero" biopsies did not show many irregularities in structural or histological construc...
In general, detection of peritoneal carcinomatosis (PC) occurs at the late stage when there is no... more In general, detection of peritoneal carcinomatosis (PC) occurs at the late stage when there is no treatment option. In the present study, we designed novel drug delivery systems that are functionalized with anti-CD133 antibodies. The C1, C2 and C3 complexes with cisplatin were introduced into nanotubes, either physically or chemically. The complexes were reacted with anti-CD133 antibody to form the labeled product of A0-o-CX-chem-CD133. Cytotoxicity screening of all the complexes was performed on CHO cells. Data showed that both C2 and C3 Pt-complexes are more cytotoxic than C1. Flow-cytometry analysis showed that nanotubes conjugated to CD133 antibody have the ability to target cells expressing the CD133 antigen which is responsible for the emergence of resistance to chemotherapy and disease recurrence. The shortest survival rate was observed in the control mice group (K3) where no hyperthermic intraperitoneal chemotherapy procedures were used. On the other hand, the longest median...
A. Immunoblot analysis of tuberin and OGG1 in different kidney regions. Different kidney regions ... more A. Immunoblot analysis of tuberin and OGG1 in different kidney regions. Different kidney regions were homogenized and protein extracts were loaded onto 7% SDS-polyacrylamide gels and transfered to PVDF membrane. The membrane was incubated with anti-tuberin or anti-OGG1 followed by different specific HRP-conjugated secondary antibodies. The proteins were visualized by ECL. GAPDH was used as loading control. B. Histograms represent means ± SE (n = 3). Significant difference from wild type rat is indicated by * P < 0.05 and ** P < 0.01.<b>Copyright information:</b>Taken from "Tuberin haploinsufficiency is associated with the loss of OGG1 in rat kidney tumors"http://www.molecular-cancer.com/content/7/1/10Molecular Cancer 2008;7():10-10.Published online 24 Jan 2008PMCID:PMC2265742.
28 Endothelial nitric oxide synthase (eNOS) is the predominant isoform that generates nitric oxid... more 28 Endothelial nitric oxide synthase (eNOS) is the predominant isoform that generates nitric oxide 29 (NO) in the blood vessels. Many different regulators govern eNOS function, including heat 30 shock protein 90 (Hsp90). Hsp90-dependent phosphorylation of eNOS is a critical event that 31 determines eNOS activity. In our earlier study, we demonstrated the occurrence of an Hsp9032 inhibitor kappa B kinase beta (IKKβ) interaction under high glucose environment. In the present 33 study, we have further defined the putative binding domain of IKKβ on Hsp90. Interestingly, 34 IKK binds to the middle domain of Hsp90, which has been shown to interact with eNOS to 35 stimulate its activity. This new finding suggests that there is a tighter regulation of eNOS activity 36 than previously assumed. Furthermore, addition of purified recombinant IKKβ to the eNOS37 Hsp90 complex reduces the eNOS-Hsp90 interaction and eNOS activity, indicating the existence 38 of a competition for Hsp90 between eNOS...
The role of tumor protein 63 (TP63) in regulating insulin receptor substrate 1 (IRS-1) and other ... more The role of tumor protein 63 (TP63) in regulating insulin receptor substrate 1 (IRS-1) and other downstream signal proteins in diabetes has not been characterized. RNAs extracted from kidneys of diabetic mice (db/db) were sequenced to identify genes that are involved in kidney complications. RNA sequence analysis showed more than 4- to 6-fold increases in TP63 expression in the diabetic mice’s kidneys, compared to wild-type mice at age 10 and 12 months old. In addition, the kidneys from diabetic mice showed significant increases in TP63 mRNA and protein expression compared to WT mice. Mouse proximal tubular cells exposed to high glucose (HG) for 48 h showed significant decreases in IRS-1 expression and increases in TP63, compared to cells grown in normal glucose (NG). When TP63 was downregulated by siRNA, significant increases in IRS-1 and activation of AMP-activated protein kinase (AMPK (p-AMPK-Th172)) occurred under NG and HG conditions. Moreover, activation of AMPK by pretreating...
Proteasome inhibitor (PI) therapy has improved the survival of multiple myeloma (MM) patients. Ho... more Proteasome inhibitor (PI) therapy has improved the survival of multiple myeloma (MM) patients. However, inevitably, primary or acquired resistance to PIs leads to disease progression; resistance mechanisms are unclear. Obesity is a risk factor for MM mortality. Oxidized LDL (OxLDL), a central mediator of atherosclerosis that is elevated in metabolic syndrome (co‐occurrence of obesity, insulin resistance, dyslipidemia and hypertension), has been linked to an increased risk of solid cancers and shown to stimulate pro‐oncogenic/survival signaling. We hypothesized that OxLDL is a mediator of chemoresistance and evaluated its effects on MM cell killing by PIs. OxLDL potently suppressed the ability of the boronic acid‐based PIs bortezomib (BTZ) and ixazomib, but not the epoxyketone‐based PI carfilzomib, to kill human MM cell lines and primary cells. OxLDL suppressed BTZ‐induced inhibition of proteasome activity and induction of pro‐apoptotic signaling. These cytoprotective effects were ab...
The role of tumor protein 63 (TP63) in regulating insulin receptor substrate 1 (IRS-1) and other ... more The role of tumor protein 63 (TP63) in regulating insulin receptor substrate 1 (IRS-1) and other downstream signal proteins in diabetes has not been characterized. RNAs extracted from kidneys of diabetic mice (db/db) were sequenced to identify genes that are involved in kidney complications. RNA sequence analysis showed more than 4- to 6-fold increases in TP63 expression in the diabetic mice’s kidneys, compared to wild-type mice at age 10 and 12 months old. In addition, the kidneys from diabetic mice showed significant increases in TP63 mRNA and protein expression compared to WT mice. Mouse proximal tubular cells exposed to high glucose (HG) for 48 h showed significant decreases in IRS-1 expression and increases in TP63, compared to cells grown in normal glucose (NG). When TP63 was downregulated by siRNA, significant increases in IRS-1 and activation of AMP-activated protein kinase (AMPK (p-AMPK-Th172)) occurred under NG and HG conditions. Moreover, activation of AMPK by pretreating...
Epithelial–mesenchymal transition (EMT) plays an important role in tissue fibrosis following chro... more Epithelial–mesenchymal transition (EMT) plays an important role in tissue fibrosis following chronic exposure to hyperglycemia. This study investigates the role of chronic diabetes in regulating tuberin/snail/AMPK to enhance EMT and increase renal fibrosis. A new mouse model of db/db/TSC2+/− was generated by backcrossing db/db mice and TSC2+/− mice. Wild type (WT), db/db, TSC2+/− and dbdb/TSC2+/− mice were sacrificed at ages 6 and 8 months old. Tuberin protein level was significantly decreased in kidneys from diabetic compared to WT mice at both ages. In addition, tuberin and E‐cadherin protein levels were significantly decreased in dbdb/TSC2+/− compared to TSC2+/− and db/db mice. In contrast, p‐PS6K, NFkB, snail, vimentin, fibronectin, and α‐SMA protein levels were significantly increased in dbdb/TSC2+/− compared to db/db and TSC2+/− mice at ages 6 and 8 months. Both downregulation of AMPK by DN‐AMPK and downregulation of tuberin by siRNA resulted in increased NFkB, snail, and fibronectin protein expression and decreased E‐cadherin protein expression in mouse primary renal proximal tubular cells. Interestingly, downregulation of snail by siRNA increased tuberin expression via feedback through activation of AMPK and reversed the expression of epithelial proteins such as E‐cadherin as well as mesenchymal proteins such as fibronectin, NF‐KB, vimentin, and α‐SMA in mouse primary renal proximal tubular cells isolated from kidneys of four mice genotypes. The data show that chronic diabetes significantly decreases tuberin expression and that provides strong evidence that tuberin is a major key protein involved in regulating EMT. These data also demonstrated a novel role for snail in regulating of AMPK/tuberin to enhance EMT and renal cell fibrosis in diabetes.
Insulin resistance is a major risk for development of type 2 diabetes. Several genes involved in ... more Insulin resistance is a major risk for development of type 2 diabetes. Several genes involved in regulation of insulin receptor substrate 1 (IRS1) and lead to chronic diabetes. Our RNA sequence data from kidney of diabetic mice show a new candidate gene involved in the regulation of IRS1. New candidate gene, Tripartite motif protein 29 (TRIM29), acts as an E3 ligase targeting insulin receptor (IR) and insulin receptor substrate 1 (IRS1) for ubiquitin-dependent degradation, resulting in insulin resistance. RNA seq data show that TRIM29 is higher 2.5 and 3.6 fold in kidney cortex of diabetic mice (db/db) at age of 10 and 12 months compared to wild type mice, respectively. Mouse renal proximal tubular epithelial cells treated with high glucose (25mM) for 48 hours showed significant increased in TRIM29 and decreased in IRS-1 compared to cells grown in normal glucose (5mM). Pre-transfected cells with siRNA against TRIM29 resulted in significant increased IRS-1 and cell proliferative mark...
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