Ectopic expression of PTTG1/securin promotes tumorigenesis in human embryonic kidney cells

BACKGROUND: Pituitary tumor transforming gene1 (PTTG1) is a novel oncogene that is expressed in most tumors. It encodes a protein that is primarily involved in the regulation of sister chromatid separation during cell division. The oncogenic potential of PTTG1 has been well characterized in the mouse, particularly mouse fibroblast (NIH3T3) cells, in which it induces cell proliferation, promotes tumor formation and angiogenesis. Human tumorigenesis is a complex and a multistep process often requiring concordant expression of a number of genes. Also due to differences between rodent and human cell biology it is difficult to extrapolate results from mouse models to humans. To determine if PTTG1 functions similarly as an oncogene in humans, we have characterized its effects on human embryonic kidney (HEK293) cells. RESULTS: We report that introduction of human PTTG1 into HEK293 cells through transfection with PTTG1 cDNA resulted in increased cell proliferation, anchorage-independent growth in soft agar, and formation of tumors after subcutaneous injection of nu/nu mice. Pathologic analysis revealed that these tumors were poorly differentiated. Both analysis of HEK293 cells transiently transfected with PTTG1 cDNA and analysis of tumors developed on injection of HEK293 cells that had been stably transfected with PTTG1 cDNA indicated significantly higher levels of secretion and expression of bFGF, VEGF and IL-8 compared to HEK293 cells transfected with pcDNA3.1 vector or uninvolved tissues collected from the mice. Mutation of the proline-rich motifs at the C-terminal of PTTG1 abolished its oncogenic properties. Mice injected with this mutated PTTG1 either did not form tumors or formed very small tumors. Taken together our results suggest that PTTG1 is a human oncogene that possesses the ability to promote tumorigenesis in human cells at least in part through the regulation of expression or secretion of bFGF, VEGF and IL-8. CONCLUSIONS: Our results demonstrate that PTTG1 is a potent human oncogene and has the ability to induce cellular transformation of human cells. Overexpression of PTTG1 in HEK293 cells leads to an increase in the secretion and expression of bFGF, VEGF and IL-8. Mutation of C-terminal proline-rich motifs abrogates the oncogenic function of PTTG1. To our knowledge, this is the first study demonstrating the importance of PTTG1 in human tumorigenesis

X-linked ichthyosis associated with psychosis and behavioral abnormalities: a case report

Abstract Background X-linked ichthyosis is a dermatological condition caused by deficiency for the enzyme steroid sulfatase. Previously, X-linked ichthyosis/steroid sulfatase deficiency has been associated with developmental and neurological phenotypes. Here, we show for the first time, that X-linked ichthyosis may be comorbid with an additional psychiatric phenotype (psychosis). Case presentation We report the case of an 11-year-old Saudi Arabian boy with X-linked ichthyosis associated with psychosis, mental retardation, autism spectrum disorder, inattentive attention deficit hyperactivity disorder, and epilepsy. Genetic analysis revealed a 1.68 Mb deletion encompassing STS in 95% of cells while biochemical analysis revealed correspondingly low steroid sulfatase activity consistent with a diagnosis of X-linked ichthyosis. The psychotic symptoms could be reasonably well controlled by administration of an atypical antipsychotic. Conclusions This report describes a case of comorbid X-linked ichthyosis and psychosis (most closely corresponding to early-onset schizophrenia) for the first time, and suggests that deficiency for steroid sulfatase and contiguous genes may increase vulnerability to psychosis as well as other psychological disorders

Cucurbitacin D ameliorates benzo[a]pyrene induced liver injury via Activation of Nrf2 Antioxidant pathway

Background: Co-morbidity variables, such as smoking, are strongly linked to the development and progression of liver cancer. Further, benzo[a]pyrene, a major component of tobacco smoke, is highly carcinogenic and triggers liver damage. Cucurbitacin, a kind of triterpene, has a wide range of biological properties, including antioxidant, anti-inflammatory, and anti-cancer effects. However, the precise mechanism of its hepatoprotective effects is obscure. Objective: The aim of this study is to investigate the cytoprotective effects of novel analog of cucurbitacin, cucurbitacin D, against benzo[a]pyrene-induced liver injury in HepG2 cells. Method: The cytoprotective efficacy of cucurbitacin D against benzo[a]pyrene-induced liver damage was studied using proliferation, clonogenicity, migration, invasion, Western blotting, and qPCR analysis. The levels of intracellular reactive oxygen species (ROS) in liver cells was measured using the DCFDA assay. Results: In human HepG2 cells, functional experiments revealed that cucurbitacin D has cytoprotective effects against dose-dependent growth inhibition by benzo[a]pyrene. The mitigation of ROS observed by fluorimeter and fluorescence microscopy suggested that this protective effect was likely due to cucurbitacin D\u27s antioxidant property. Additional research is ongoing to identify the effect of cucurbitacin D on oxidative stress markers by using qPCR and western blotting techniques. Overall, these findings showed that cucurbitacin D diminishes benzo[a]pyrene-induced liver injury via its antioxidant activity. Conclusion: These findings show that cucurbitacin D has hepatoprotective properties against benzo[a]pyrene-induced liver injury, making it an attractive food supplement ingredient

Hepatoprotective role of Cucurbitacin D on benzo[a]pyrene induced liver injury

Background: Epidemiological findings show the strong correlation of co-morbidity factors including smoking with the development and progression of liver cancer. Moreover, benzo[a]pyrene, a main component of tobacco smoke, is extremely carcinogenic and contributes to liver injury as well. Cucurbitacin, chemically classified as triterpenes, have shown diverse biological activities including potent antioxidant, anti-inflammatory and anti-cancer activities. However, their hepatoprotective activities are not completely understood. Objective: In the present study, we investigated the cytoprotective activity of novel analog of cucurbitacin, cucurbitacin D, against benzo[a]pyrene-induced liver injury in human HepG2 cells. Method: Proliferation, clonogenicity, migration, invasion, Western blotting and qPCR analysis were conducted to investigate the cytoprotective effect of cucurbitacin D against benzo[a]pyrene induced liver damage. DCFDA assay was performed to analyze intracellular reactive oxygen species (ROS) level in liver cells. Results: Functional assays showed that cucurbitacin D exhibited cytoprotective effects against dose-dependent growth inhibition by benzo[a]pyrene in human HepG2 cells. This protective effect was likely associated with antioxidant potential of cucurbitacin D, as evidenced by the attenuation of ROS observed by fluorimeter and fluorescence microscopy. Further study is ongoing to examine the effect of cucurbitacin D on oxidative stress markers by employing western blotting and qPCR techniques. Collectively, these results exhibited that cucurbitacin D alleviate benzo[a]pyreneinduced liver injury through its antioxidant effects. Conclusion: These results have demonstrated hepatoprotective effects of cucurbitacin D against benzo[a]pyrene-induced liver damage, rendering it as an effective potential ingredient in food supplements

Therapeutic efficacy of ormeloxifene against hepatocellular carcinoma

Background: Hepatocellular carcinoma (HCC) is the most common primary liver cancer and leading cause of cancer related deaths worldwide. Severe toxicity and drug resistance to available chemotherapeutic agents display ineffective clinical response. Therefore, drug repurposing is gaining attention owing to their known biological activities and excellent safety profiles. Ormeloxifene (ORM), non-steroidal, selective estrogen receptor modulator (SERM), and exhibit diverse pharmacological activities. The aim of this study is to assess the therapeutic activity of ORM and to investigate the underlying molecular mechanism against hepatocellular carcinoma. Objective: To investigate the therapeutic activity of ormeloxifene in human hepatocellular carcinoma cells. Methodology: MTT and colony formation assays were performed in SK-Hep-1, Hep3B and C3A cells. In vitro functional assays were carried out for investigating effect of ORM on migration and invasion abilities of HCC cells using Boyden chamber and Matrigel assays respectively. Results: Functional analysis revealed that ORM treatment led to suppression of proliferation and colony formation in human hepatocellular carcinoma cells in dose and time-dependent manner compared to vehicle treated group. ORM treatment, as shown by wound healing and Matrigel invasion assay, respectively, suppresses the migration and invasion of human hepatocellular carcinoma cells. Further, experiments are underway to determine the effect of ORM on EMT markers using western blotting and qPCR techniques. Conclusion: Taken together, ORM exhibited potent anticancer effects against HCC and could be further explored as a novel therapeutic modality for the treatment of HCC

Pharmacological restoration of PKD1: A novel strategy for prostate cancer therapy

Background: Prostate cancer has poor prognosis owing to late diagnosis and ineffective multimodal clinical treatment. Extensive efforts are ongoing to establish methods that can resolve the expression of genes implicated in disease development and treatment. Previously, we reported that Protein Kinase D1 (PKD1), a serine threonine kinase, controls a number of tumor suppressor functions including cell aggregation, cell motility, cell proliferation, and cell invasion. Thus, PKD1 is considered as an emerging therapeutic target for prostate cancer treatment. Objective: To investigate the restoration of PKD1 by a pharmacological modulator ormeloxifene, which showed well-defined PK/PD and safety profiles in humans. Methods: Proliferation, clonogenicity, migration, invasion, western blotting and qPCR analysis were performed to investigate the anticancer effect of ORM, docetaxel and/or their combination on PKD1 and related signaling mechanisms in prostate cancer. Results: ORM treatment inhibited cell proliferation, invasion, migration and colony formation abilities of prostate cancer cells in a dose-dependent manner compared to vehicle treated group. ORM treatment selectively induces the expression of PKD1 both at mRNA and protein levels in C4-2 cells. Moreover, our results have also shown that ORM effectively attenuates MTA1 expression in prostate cancer cells. MTA1 physically interact and shown to have inverse relationship with PKD1. In addition, we observed that ORM treatment enhances the therapeutic efficacy of docetaxel in C4-2 cells. Our results also indicate that ORM treatment potentiate the effects of docetaxel as determined by MTS and colony formation assays. Conclusion: These results suggest that ORM exhibit potent anticancer activity via restoration of PKD1 in prostate cancer

Molecular Insights into Targeting PKD1 for Prostate Cancer Treatment

Background: Prostate cancer has a poor prognosis due to late diagnosis and ineffective multimodal clinical treatment. Efforts are underway to create strategies for resolving the abnormal expression of molecular targets implicated in disease development and progression. We previously reported that the serine threonine kinase Protein Kinase D1 (PKD1) regulates a multitude of tumor suppressor functions, including cell aggregation, motility, proliferation, and invasion in prostate cancer. Thus, PKD1 is regarded as a promising therapeutic target for the treatment of prostate cancer. Objective: The goal of this study was to investigate the therapeutic potential of ormeloxifene (ORM), a pharmacological modulator with well-defined PK/PD and safety profiles in humans, for PKD1 restoration in prostate cancer. Methods: The anticancer effect of ORM on PKD1 and associated signaling mechanisms in prostate cancer was investigated using proliferation, clonogenicity, migration, invasion, western blotting, and qPCR analysis. Results: In comparison to the vehicle-treated group, ORM treatment decreased prostate cancer cell proliferation, invasion, migration, and colony formation in a dose-dependent manner. In C4-2 cells, ORM treatment selectively induces PKD1 expression at both the mRNA and protein levels. Furthermore, our findings revealed that ORM efficiently suppresses MTA1 expression in prostate cancer cells. MTA1 physically interacts with PKD1 and has been shown to have an inverse correlation with it. Our results also showed that ORM treatment enhances the therapeutic efficacy of decetaxel. Conclusion: Taken together, these findings show that ORM has anticancer properties in prostate cancer via restoring PKD1

Cucurbitacin D exhibits potent anticancer activity in cervical cancer

In this study, we for the first time, investigated the potential anti-cancer effects of a novel analogue of cucurbitacin (Cucurbitacin D) against cervical cancer in vitro and in vivo. Cucurbitacin D inhibited viability and growth of cervical cancer cells (CaSki and SiHa) in a dose-dependent manner. IC50 of Cucurbitacin D was recorded at 400 nM and 250 nM in CaSki and SiHa cells, respectively. Induction of apoptosis was observed in Cucurbitacin D treated cervical cancer cells as measured by enhanced Annexin V staining and cleavage in PARP protein. Cucurbitacin D treatment of cervical cancer cells arrested the cell cycle in G1/S phase, inhibited constitutive expression of E6, Cyclin D1, CDK4, pRb, and Rb and induced the protein levels of p21 and p27. Cucurbitacin D also inhibited phosphorylation of STAT3 at Ser727 and Tyr705 residues as well as its downstream target genes c-Myc, and MMP9. Cucurbitacin D enhanced the expression of tumor suppressor microRNAs (miR-145, miRNA-143, and miRNA34a) in cervical cancer cells. Cucurbitacin D treatment (1 mg/kg body weight) effectively inhibited growth of cervical cancer cells derived orthotopic xenograft tumors in athymic nude mice. These results demonstrate the potential therapeutic efficacy of Cucurbitacin D against cervical cancer

How Does The Fasting of Ramadan Affect Breast Milk Constituents?

Background: Breast-feeding of infants is associated with their better biological, psychological and intellectual development. However, many factors affect the volume and composition of human milk such as stage of lactation and maternal diet. Many breast-feeding Muslim mothers fast the lunar month of Ramadan. The effects of fasting on milk constituents have not been previously studied in Sudan. Therefore, we aimed to investigate the variations between milk constituents during fasting and non-fasting periods among a group of Sudanese women.Materials and Methods: Twenty four healthy breast-feeding mothers volunteered to participate in this cross-sectional study. Each mother provided 100 ml of breast milk during fasting and again 2 weeks after end of the fasting month of Ramadan. Milk was properly stored and analyzed for the various constituents, using the appropriate laboratory methods. The main constituents analyzed were: ash, protein, lactose, iron and electrolytes.Results: The age range of lactating women was between 18 and 38 years, mean (+SD) 28.8 (± 5.15 years). Most mothers 17 (70.8%) were house-wives. Analysis of breast milk during fasting and non-fasting periods showed that: lactose, protein, sodium, potassium, calcium and phosphate were significantly decreased in the fasting breast milk compared with the non-fasting milk (p=0.01), while total soluble solid, moisture, ash and iron constituents had not significantly changed during fasting.Conclusion: Fasting of Ramdan significantly affects proteins, carbohydrates and electrolytes in breast milk.Keywords: breast-feeding, milk constituents, Ramadan, fasting

Novel Mechanistic Insight into the Anticancer Activity of Cucurbitacin D against Pancreatic Cancer (Cuc D Attenuates Pancreatic Cancer)

Pancreatic cancer (PanCa) is one of the leading causes of death from cancer in the United States. The current standard treatment for pancreatic cancer is gemcitabine, but its success is poor due to the emergence of drug resistance. Natural products have been widely investigated as potential candidates in cancer therapies, and cucurbitacin D (Cuc D) has shown excellent anticancer properties in various models. However, there is no report on the therapeutic effect of Cuc D in PanCa. In the present study, we investigated the effects of the Cuc D on PanCa cells in vitro and in vivo. Cuc D inhibited the viability of PanCa cells in a dose and time dependent manner, as evident by MTS assays. Furthermore, Cuc D treatment suppressed the colony formation, arrest cell cycle, and decreased the invasion and migration of PanCa cells. Notably, our findings suggest that mucin 13 (MUC13) is down-regulated upon Cuc D treatment, as demonstrated by Western blot and qPCR analyses. Furthermore, we report that the treatment with Cuc D restores miR-145 expression in PanCa cells/tissues. Cuc D treatment suppresses the proliferation of gemcitabine resistant PanCa cells and inhibits RRM1/2 expression. Treatment with Cuc D effectively inhibited the growth of xenograft tumors. Taken together, Cuc D could be utilized as a novel therapeutic agents for the treatment/sensitization of PanCa