Sita Aggarwal - Academia.edu (original) (raw)

Papers by Sita Aggarwal

The abbreviations used are: EMSA, electrophoretic mobility shift assay; EGFR, epidermal growth fa... more The abbreviations used are: EMSA, electrophoretic mobility shift assay; EGFR, epidermal growth factor receptor; HER, human epidermal growth factor receptor, IAP, inhibitor-of-apoptosis protein; IκB, inhibitory subunit of NF-κB; FLICE, Fas associated death domain protein-like interleukin-1β-converting enzyme inhibitory protein; FLIP, FLICE-inhibitory protein; iNOS, inducible nitric oxide synthase; MMP, matrix metalloproteinase; NF-κB, nuclear factor-κB; NIK, NF-κB-inducing kinase; PBS, phosphate-buffered saline; SDS, sodium dodecyl sulfate; SEAP, secretory alkaline phosphatase; VEGF, vascular endothelial growth factor, XIAP, X-chromosome-linked IAP.

Suppression of active CREB induces apoptosis and inhibition of cell growth in human non-small-cell Lung cancer cells

Cancer Research, 2006

2625 Genes that are regulated by cyclicAMP response element binding (CREB) protein suppress apopt... more 2625 Genes that are regulated by cyclicAMP response element binding (CREB) protein suppress apoptosis, induce cell proliferation, and mediate inflammation, angiogenesis and tumor metastasis. It was not known whether CREB is involved in lung cancer. We hypothesized that constitutively active CREB is an important target in the treatment of non-small cell lung cancer (NSCLC) and that its inhibition should block cell proliferation and induce apoptosis in NSCLC cells. We found that the NSCLC cell lines we examined overexpressed constitutively active CREB, two of its upstream kinases, phospho- ribosomal s6-kinase (Rsk), phospho-extracellular signal kinase(Erk1/2) and cell survival genes such as Bcl-2 and Bcl-XL. Ectopic expression of a dominant-repressor CREB (KCREB), mutant CREB (CREB133, Ser133Ala), and small interfering (si) RNA against CREB (siCREB) suppressed the cell proliferation of NSCLC (H1734) cells. Furthermore, treatment of H1734 cells with a Protein Kinase C inhibitor (Ro-31-...

Targeting protein kinase C beta enhances cell death of non-small cell lung cancer cells

Cancer Epidemiology and Prevention Biomarkers, 2006

Fifth AACR International Conference on Frontiers in Cancer Prevention Research, Nov 12-15, 2006 A... more Fifth AACR International Conference on Frontiers in Cancer Prevention Research, Nov 12-15, 2006 A127 The protein kinase C (PKC) family consists of structurally related serine-threonine kinases that influence drug resistance and cell survival in various cancers. In this study we identified that two of the classical PKC isoforms, PKC-α and PKC-β that played an important role in cell survival of non-small cell lung cancer (NSCLC) cells. Various NSCLC cells (H1734, A549, H226, H1703, H292) expressed high levels of phosphorylated PKC-α, PKC-β and PKC-μ when compared to normal human tracheobronchial epithelial (NHTBE) cells. The levels of phosphorylated PKC-δ or PKC-ζ, ι/λ were reduced in NSCLC when compared to NHTBE. Knock down of protein kinase expression by small interfering (si) RNAs, siPKCβ alone or by combined siPKCα/β and to a lesser extent by siPKCα alone inhibited the expression of the cAMP response element (CRE)-binding protein (CREB) transcription factor in H1734 NSCLC cell lin...

Chemico-Biological Interactions, 2020

Juglone and thymoquinone are cytotoxic to pancreatic cancer cells. The aim of this study was to i... more Juglone and thymoquinone are cytotoxic to pancreatic cancer cells. The aim of this study was to investigate, using an analysis of isobolograms, the type and degree of interactions between juglone and thymoquinone on MIA PaCa-2 pancreatic cancer cells. Cell viability was evaluated using the MTT assay. Cell death was determined by flow cytometry. The IC 50 value for juglone and TQ in combination was found to be 24.75 μM, which was higher than juglone or TQ alone. Juglone alone killed Mia Paca-2 cells by ferroptosis. At concentrations where 10, 20 or 50% of cells were affected, there existed a moderate antagonistic relationship between juglone and TQ as indicated by the combination index (CI) value determined by the Compusyn software. At concentrations that affected 75% and 90% of cells, there were nearly an additive effect with CI value of 1.09249 and 0.92391, respectively. Moderate synergism was only seen at concentration where 95% of cells were affected, and the corresponding concentration of juglone and TQ at that combination was 40.90 μM and 511.19 μM, respectively.

Food Production, Processing and Nutrition, 2020

Glucolipotocixity induces IL-1 β secretion which impairs pancreatic β-cell insulin secretion. Ell... more Glucolipotocixity induces IL-1 β secretion which impairs pancreatic β-cell insulin secretion. Ellagic acid and urolithin A have strong anti-inflammatory effect on cells. Muscadine and amla are very good sources of ellagic acid. The present study examined the effect of ellagic acid, ellagic acid-rich muscadine or amla extract, or urolothin A on inflammation in β cells under glucolipotoxic conditions. Rat NIT-1 β cells were incubated in glucolipotoxic conditions (33.3 mM glucose, 250 μM palmitic acid or 33.3 mM glucose + 250 μM palmitic acid with or without ellagic acid, ellagic acid-rich muscadine or amla extracts standardized to its ellagic acid content, or urolithin A). Inflammatory status was evidenced by ELISA analysis of insulin and IL-1β secretion. Ellagic acid-rich muscadine or amla extracts dose-dependently stimulated insulin secretion and down-regulated IL-1β better than pure ellagic acid, or urolithin A. Urolithin A did not statistically stimulate insulin secretion and did ...

Abstract 4128: Anti-angiogenic and anti-metastatic activities of juglone in pancreatic cancer cells

Cancer Research, 2016

Pancreatic cancer is the fourth leading cause of cancer related deaths in the US. The late diagno... more Pancreatic cancer is the fourth leading cause of cancer related deaths in the US. The late diagnosis, early metastasis and poor survival rate make this disease lethal. This current study was designed to examine the effect of juglone, 5 hydroxy-1, 4-naphthoquinone in modulating the angiogenic and metastatic potential of pancreatic cancer cells in vitro, using the MIA Paca-2 human pancreatic cancer cell line. Juglone is a natural quinoid abundantly found abundantly in plants of Juglandacea family. To understand the anti-angiogenic and anti-metastatic properties of juglone, expression of markers related to angiogenesis were measured by Western blot or ELISA and the effect on ability of cancer cells to migrate and invade after treatment was analyzed by transwell invasion and migration assay, wound healing assay, and in vitro tube formation assay in human umbilical vein endothelial cells. The Akt pathway is highly upregulated and is responsible for the regulation of various biological pr...

Journal of Food Bioactives, 2018

Hypoxia-inducible factor -1 alpha (HIF-1α), vascular endothelial growth factor (VEGF), and phosph... more Hypoxia-inducible factor -1 alpha (HIF-1α), vascular endothelial growth factor (VEGF), and phosphorylated Akt (p-Akt) are critical in pancreatic cancer cell growth, angiogenesis and metastasis. The ability of MIA Paca-2 pancreatic cancer cells to migrate and invade after treatment with juglone (5 hydroxy-1, 4-naphthoquinone) was analyzed by a transwell invasion and migration assay, a wound healing assay, and an in vitro tube formation assay using human umbilical vein endothelial cells (HUVEC). ELISA was used to determine the levels of HIF1-α. Western blot analysis was used to determine the level of VEGF, p-Akt, and carbonic anhydrase IX expression. Juglone down-regulated the expression of HIF-1α, VEGF, and significantly suppressed the phosphorylation of Akt. Juglone dose-dependently inhibited the metastatic potential of pancreatic cancer cells by inhibiting cell migration, invasion and wound healing assays. Juglone attenuated the aggressiveness of pancreatic cancer cells in vitro.

The mouse dead-end gene isoform a is necessary for germ cell and embryonic viability

Biochem Biophys Res Commun, 2007

Curcumin Conjugates for Treating and Preventing Cancers

Abstract B56: Inactivation of LKB1-AMPK pathway is mediated by inflammation in pancreatic cancer

Cancer Research, 2015

Goal: The goal of this project is to determine if there is link between cancer cell metabolism an... more Goal: The goal of this project is to determine if there is link between cancer cell metabolism and inflammation. LKB1is a known serine/threonine kinase tumor suppressor gene and it functions by sensing changes in cellular energy homeostasis and modulates anabolic and catabolic processes by activating its downstream kinase, AMP-activated protein kinase (AMPK). It is associated with Peutz-Jeghers syndrome, as well as various cancers including pancreatic cancer. Thus; it is a unique link between cell metabolism and inflammation. Hypothesis: Our hypothesis is that LKB1-AMK succumbs to inactivation processes because of increased NF-κB expression in pancreatic cancer cells and is associated with increased pancreatic cancer growth. Results: We tested five pancreatic cancer cells that express constitutive active NF-κB (pp65) and LKB1proteins by western blot analysis. In these cancer cells there was loss of active AMPK, phospho AMK (pAMPK) protein and its substrate pACC protein, suggesting a...

This information is current as by TNF Induced Activation and Suppresses Apoptosis B κ Gene-7/IL-2... more This information is current as by TNF Induced Activation and Suppresses Apoptosis B κ Gene-7/IL-24 Gene Enhances NF-Melanoma Differentiation-Associated

Studies demonstrate that flexor tendons contain loosely associated biomolecules which inhibit its... more Studies demonstrate that flexor tendons contain loosely associated biomolecules which inhibit its mineralization under physiological conditions. Based upon their molecular weights, these inhibitory biomolecules, could be classified into two categories, having molecular weights less than and greater than 13,000 daltons. The main inhibitory biomolecule was found to be an acidic polypeptide having molecular weight of 12,400 daltons.

International Journal of Obesity, 2012

Background-Cellular glucose uptake can be enhanced by upregulating Ras signaling in either insuli... more Background-Cellular glucose uptake can be enhanced by upregulating Ras signaling in either insulin-dependent or-independent manner. In presence of insulin and intact insulin signaling, Ras has a negligible role in glucose uptake. Conversely, when insulin signaling is impaired in obesity or diabetes, the insulin-independent Ras pathway may be valuable for enhancing glucose disposal. We previously reported that Ad36, a human adenovirus, enhances cellular glucose uptake by upregulating the Ras/Glut4 pathway. Here, we investigated if Ad36-upregulated Ras via the insulin-independent pathway, to enhance glucose uptake. Furthermore, uncontrolled upregulation of Ras is linked with oncogenic cell transformation, if the tumor-suppressor gene p53 is also downregulated. Hence, we determined if upregulation of Ras by Ad36 would induce oncogenic cell transformation. Finally, we determined the relevance of Ad36 to insulin resistance in humans. Methods-Insulin receptor (IR) was knocked down with small interfering RNA in 3T3-L1 adipocytes, to determine if Ad36 increases the Ras/Glut4 pathway and glucose uptake without IRsignaling. Next, the effects of Ad36 on cell transformation and p53 abundance were determined. Finally, overweight or obese women were screened for seropositivity to Ad36, as an indicator of natural Ad36 infection. Associations of Ad36 infection with adiposity and C-reactive proteins (CRPs)-two key markers of insulin resistance, and with glucose disposal, were determined. Results-Unaffected by IR knock-down, Ad36 significantly increased the Ras pathway, Glut4 translocation and glucose uptake in 3T3-L1 adipocytes. Despite Ras upregulation, Ad36 did not transform 3T3-L1 cells. This may be because Ad36 significantly increased p53 protein in 3T3-L1

Laser capture microdissection of pancreatic ductal adeno-carcinoma cells to analyze EzH2 by Western Blot analysis

Methods in molecular biology (Clifton, N.J.), 2011

Pure populations of tumor cells are essential for the identification of tumor-associated proteins... more Pure populations of tumor cells are essential for the identification of tumor-associated proteins for the development of targeted therapy. In recent years, laser capture microdissection (LCM) has been used successfully to obtain distinct populations of cells for subsequent molecular analysis. The polycomb group (PcG) protein, enhancer of zeste homolog 2 (EzH2), a methyl-transferase that plays a key role in -transcriptional gene repression, is frequently overexpressed in several malignant tumors. High levels of EzH2 are often associated with advanced disease stage in many solid tumors; however, its role in the pathogenesis of pancreatic ductal adeno-carcinoma (PDAC) is poorly understood. Because of the limited sample availability and the absence of in vitro amplification steps for proteins, the use of LCM for proteomics studies largely depends on highly sensitive protein detection methods. Here, we developed a faster and sensitive Western blot protocol and validated it for the detect...

Indian journal of biochemistry & biophysics, 2005

The role of biomolecule(s) from renal stone matrix in urolithiasis was investigated. The ability ... more The role of biomolecule(s) from renal stone matrix in urolithiasis was investigated. The ability of a particular fraction (> 10 kDa fraction) isolated from the EDTA extract of powdered human renal stones to influence calcium oxalate monohydrate (COM) crystal growth was studied. The most potent inhibitor of COM crystal growth obtained from > 10 kDa fraction was purified by various chromatographic techniques and SDS-PAGE, etc. and was found to have a molecular mass of 36 kDa. The urine and serum samples obtained from normal persons were found to be more potent in inhibiting the growth of COM crystals as compared to the kidney-stone patients. Polyclonal antibodies were raised against this inhibitor and were employed to determine the concentration of 36 kDa inhibitor in urine and serum samples of normal persons and kidney-stone patients.

IL-24 in Regulation of Antitumor Immune Response and in Signaling

Targeted Cancer Immune Therapy, 2009

ABSTRACT In cancer therapy, cytokines are generally used to increase immunity. Cytokines are eith... more ABSTRACT In cancer therapy, cytokines are generally used to increase immunity. Cytokines are either proteins or glycoproteins, which are secreted by immune cells. It is now known that the tumor microenvironment secretes a mixture of cytokines that plays an important role in carcinogenesis. In chronic inflammation, cytokines released at the site of a tumor facilitate tumor growth, instead of promoting antitumor immunity. In 1995, a protein called melanoma differentiation-associated gene-7 (MDA-7), also known as suppressor of tumorigenicity-16 (ST-16), was discovered; it was renamed as interlukine-24 because of its cytokine-like properties and was found to inhibit the growth and proliferation in melanoma cells. The rat counterpart of IL-24 was named as mob-5 or C49a. The murine counterpart was named as FISP. Ectopic expression of MDA-7/IL-24 by means of a replication defective adenovirus (Ad-MDA-7/IL-24) results in growth suppression, inhibits angiogenesis and apoptosis not only in melanoma cells but also in numerous other cancer cell types such as glioblastoma, carcinomas of breast, colon, lung, ovarian and prostate sparing normal epithelial and fibroblasts. Therefore, this article reviews anti-cytokine therapies of MDA-7/IL-24 being pursued in cancer and more details about signaling pathways associated with MDA-7/IL-24 in cancer.

Abstract 5600: Destruction of prostate cancer cell xenografts by FSH-Lytic peptide conjugates

Cancer Research, 2013

Background: In previous studies (Hansel, et al., Mol. and Cell. Endocrinol. 269:26-33, 2007),we s... more Background: In previous studies (Hansel, et al., Mol. and Cell. Endocrinol. 269:26-33, 2007),we showed that conjugates of membrane destroying lytic peptides with either LHRH or with a 15-amino acid segment of the β chain of human chorionic gonadotropin (hCG) target and destroy human prostate, breast and ovarian cancer cells in tumor bearing nude mice. Recently (Radu et al., N. England J. Med. 363:1621-1630, 2012) reported that endothelial cells in blood vessels supplying cancers express follicle-stimulating hormone (FSH) receptors. Objectives:The objectives of this study were to synthesize a bioconjugate of a lyticpeptide (Phor18) to each of three segments of the β chain of FSH that are known to bind to the FSH receptor, and test these conjugates (FSH90-95-Phor18, FSH81-95-Phor18 and FSH33-53-Phor18) for their ability to target and lyse prostate cancer cells in vitro and in vivo. Results: In in vivo experiments, administration of FSH90-95-Phor18 and FSH81-95-Phor18 significantly (p ...

PLoS ONE, 2008

Background: The dead-end (Dnd1) gene is essential for maintaining the viability of germ cells. In... more Background: The dead-end (Dnd1) gene is essential for maintaining the viability of germ cells. Inactivation of Dnd1 results in sterility and testicular tumors. The Dnd1 encoded protein, DND1, is able to bind to the 39-untranslated region (UTR) of messenger RNAs (mRNAs) to displace micro-RNA (miRNA) interaction with mRNA. Thus, one function of DND1 is to prevent miRNA mediated repression of mRNA. We report that DND1 interacts specifically with APOBEC3. APOBEC3 is a multifunctional protein. It inhibits retroviral replication. In addition, recent studies show that APOBEC3 interacts with cellular RNA-binding proteins and to mRNA to inhibit miRNA-mediated repression of mRNA. Methodology/Principal Findings: Here we show that DND1 specifically interacts with another cellular protein, APOBEC3. We present our data which shows that DND1 co-immunoprecipitates APOBEC3 from mammalian cells and also endogenous APOBEC3 from mouse gonads. Whether the two proteins interact directly remains to be elucidated. We show that both DND1 and APOBEC3 are expressed in germ cells and in the early gonads of mouse embryo. Expression of fluorescentlytagged DND1 and APOBEC3 indicate they localize to the cytoplasm and when DND1 and APOBEC3 are expressed together in cells, they sequester near peri-nuclear sites. Conclusions/Significance: The 39-UTR of mRNAs generally encode multiple miRNA binding sites as well as binding sites for a variety of RNA binding proteins. In light of our findings of DND1-APOBEC3 interaction and taking into consideration reports that DND1 and APOBEC3 bind to mRNA to inhibit miRNA mediated repression, our studies implicate a possible role of DND1-APOBEC3 interaction in modulating miRNA-mediated mRNA repression. The interaction of DND1 and APOBEC3 could be one mechanism for maintaining viability of germ cells and for preventing germ cell tumor development.

An extract of artemisia drancunculus L. stimulates insulin secretion from β cells, activates AMPK and suppresses inflammation

Pancreatology, 2013

ABSTRACT Artemisia dracunculus L. (Russian tarragon) is a perennial herb belonging to the family ... more ABSTRACT Artemisia dracunculus L. (Russian tarragon) is a perennial herb belonging to the family Compositae and has a history of medicinal use in humans, particularly for treatment of diabetes. In this study a defined plant extract from Artemisia dracunculus L. (termed PMI-5011) is used to improve β cells function and maintain β cell number in pancreatic islets as an alternative drug approach for successful treatment of diabetes. Mouse and human pancreatic beta cells were treated with defined plant extract of Artemisia dracunculus L. (PMI-5011) to understand the mechanism(s) that influence beta cell function and β cell number. We found that the PMI-5011 enhances insulin release from primary β cells, isolated mouse and human islets and it maintains β cell number. Insulin released by PMI-5011 is associated with the activation of AMP-activated protein kinase (AMPK), and protein kinase B (PKB). Furthermore, PMI-5011 suppresses LPS/INFγ-induced inflammation and inflammatory mediator(s) in macrophages. PMI-5011 inhibited Nitric oxide (NO) production and expression of inducible nitric oxide synthase (iNOS) at the protein level and also attenuated pro-inflammatory cytokine (IL-6) production in macrophages. PMI-5011 has potential therapeutic value for diabetes treatment via increasing insulin release from β cells and decreases capacity of macrophages to combat inflammation. Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.

Molecular and Cellular Biochemistry, 2000

Human renal calculi surgically removed from kidney stone patients were obtained and chemically an... more Human renal calculi surgically removed from kidney stone patients were obtained and chemically analysed. Stones with CaOx (calcium oxalate) as the major component were washed in 0.15 M NaCl with gentle stirring for 48 h and then pulverised to a fine powder. The powder was extracted with 0.05 M EGTA, 1 mM PMSF and 1% ß- mercaptoethanol for 4 days