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ABSTRACT: Planar triazinium cationic species from vanadyl-assisted cyclization of 1-(2-thiazolylazo)-2-naphthol (H-TAN, ), 1-(2-pyridylazo)-2-naphthol (H-PAN, ), 2-(2'-thiazolylazo)-p-cresol (H-TAC, ) and 6-(2'-thiazolylazo)-resorcinol (H-TAR, ) were prepared and characterized. A dioxovanadium(v) species [VO(2)(TAR)] () was also isolated. Compounds , and were structurally characterized. Both and have planar structures. Complex has V(V)O(3)N(2) coordination geometry. The cyclised triazinium compound forms a radical species within -0.06 to -0.29 V vs. SCE in DMF-0.1 M tetrabutylammonium perchlorate with a second response due to formation of an anionic species. A confocal microscopic study showed higher nuclear uptake for having a fused thiazole moiety than with a fused pyridine ring. The compounds showed a partial intercalative mode of binding to calf thymus DNA. Compound showed plasmid DNA photo-cleavage activity under argon and photocytotoxicity in HeLa and MCF-7 cells with IC(50) values of 15.1 and 3.4 μM respectively in visible light of 400-700 nm, while being essentially non-toxic in the dark with IC(50) values of 90.4 and 21.9 μM. A TDDFT study was done to rationalize the experimental data.
Dalton Transactions 01/2013; · 3.84 Impact Factor
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Arivazhagan Arimappamagan,
Kumaravel Somasundaram,
Kandavel Thennarasu,
Sreekanthreddy Peddagangannagari,
Harish Srinivasan,
Bangalore C Shailaja,
Cini Samuel,
Irene Rosita Pia Patric,
Sudhanshu Shukla,
Balaram Thota,
Krishnarao Venkatesh Prasanna,
Paritosh Pandey,
Anandh Balasubramaniam,
Vani Santosh,
Bangalore Ashwathnarayanara Chandramouli,
Alangar Sathyaranjandas Hegde, Paturu Kondaiah,
Manchanahalli R Sathyanarayana Rao
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ABSTRACT: Recent research on glioblastoma (GBM) has focused on deducing gene signatures predicting prognosis. The present study evaluated the mRNA expression of selected genes and correlated with outcome to arrive at a prognostic gene signature.
Patients with GBM (n = 123) were prospectively recruited, treated with a uniform protocol and followed up. Expression of 175 genes in GBM tissue was determined using qRT-PCR. A supervised principal component analysis followed by derivation of gene signature was performed. Independent validation of the signature was done using TCGA data. Gene Ontology and KEGG pathway analysis was carried out among patients from TCGA cohort.
A 14 gene signature was identified that predicted outcome in GBM. A weighted gene (WG) score was found to be an independent predictor of survival in multivariate analysis in the present cohort (HR = 2(.)507; B = 0(.)919; p<0(.)001) and in TCGA cohort. Risk stratification by standardized WG score classified patients into low and high risk predicting survival both in our cohort (p = <0(.)001) and TCGA cohort (p = 0(.)001). Pathway analysis using the most differentially regulated genes (n = 76) between the low and high risk groups revealed association of activated inflammatory/immune response pathways and mesenchymal subtype in the high risk group.
We have identified a 14 gene expression signature that can predict survival in GBM patients. A network analysis revealed activation of inflammatory response pathway specifically in high risk group. These findings may have implications in understanding of gliomagenesis, development of targeted therapies and selection of high risk cancer patients for alternate adjuvant therapies.
PLoS ONE 01/2013; 8(4):e62042. · 4.09 Impact Factor
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ABSTRACT: Oral submucous fibrosis (OSF) is a chronic inflammatory disease characterized by the accumulation of excess collagen, and areca nut chewing has been proposed as an important etiological factor for disease manifestation. Activation of transforming growth factor-b signaling has been postulated as the main causative event for increased collagen production in OSF. Oral epithelium plays important roles in OSF, and arecoline has been shown to induce TGF-b in epithelial cells. In an attempt to understand the role of areca nut constituents in the manifestation of OSF, we studied the global gene expression profile in epithelial cells (HaCaT) following treatment with areca nut water extract or TGF-b. Interestingly, 64% of the differentially regulated genes by areca nut water extract matches with the TGF-b induced gene expression profile. Out of these, expression of 57% of genes was compromised in the presence of ALK5 (TbRI) inhibitor and 7% were independently induced by areca nut, highlighting the importance of TGF-b in areca nut actions. Areca nut water extract treatment induced p-SMAD2 and TGF-b downstream targets in HaCaT cells but not in human gingival fibroblast cells (hGF), suggesting epithelial cells could be the source of TGF-b in promoting OSF. Water extract of areca nut consists of polyphenols and alkaloids. Both polyphenol and alkaloid fractions of areca nut were able to induce TGF-b signaling and its downstream targets. Also, SMAD-2 was phosphorylated following treatment of HaCaT cells by Catechin, Tannin and alkaloids namely Arecoline, Arecaidine and Guvacine. Moreover, both polyphenols and alkaloids induced TGF-b2 and THBS1 (activator of latent TGF-b) in HaCaT cells suggesting areca nut mediated activation of p-SMAD2 involves up-regulation and activation of TGF-b. These data suggest a major causative role for TGF-b that is induced by areca nut in OSF progression.
PLoS ONE 12/2012; · 4.09 Impact Factor
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ABSTRACT: Lipoplexes formed by the pEGFP-C3 plasmid DNA (pDNA) and lipid mixtures containing cationic gemini surfactant of the 1, 2-bis(hexadecyl dimethyl ammonium) alkanes family referred to as C(16)C(n)C(16), where n = 2, 3, 5, or 12, and the zwitterionic helper lipid, 1, 2-dioleoyl-sn-glycero-3-phosphatidylethanolamine (DOPE) have been studied from a wide variety of physical, chemical and biological standpoints. The study has been carried out using several experimental methods, such as zeta potential, gel electrophoresis, small-angle X-ray scattering (SAXS), cryo-TEM, gene transfection, cell viability/cytotoxicity and confocal fluorescence microscopy. As reported recently in a communication (J. Am. Chem. Soc. 2011, 133, 18014), the detailed physicochemical and biological studies confirm that, in the presence of the studied series lipid mixtures, plasmid DNA is compacted with a large number of its associated Na(+) counterions. This in turn yields a much lower effective negative charge, q(pDNA), a value that has been experimentally obtained for each mixed lipid mixture. Consequently, the cationic lipid (CL) complexes prepared with pDNA and CL/DOPE mixtures to be used in gene transfection require significantly less amount of CL than the one estimated assuming a value of q(DNA)=-2. This drives to a considerably lower cytotoxicity of the gene vector. Depending on the CL molar composition, α, of the lipid mixture, and the effective charge ratio of the lipoplex, ρ(eff), the reported SAXS data indicate the presence of two or three structures in the same lipoplex, one in the DOPE rich region, other in the CL rich region, and another one present at any CL composition. Cryo-TEM and SAXS studies with C(16)C(n)C(16)/DOPE-pDNA lipoplexes indicate that pDNA is localized between the mixed lipid bilayers of lamellar structures within a monolayer of ~2 nm. This is consistent with a highly compacted supercoiled pDNA conformation compared with that of linear DNA. Transfection studies were carried out with HEK293T, HeLa, CHO, U343 and H460 cells. The α and ρ(eff) values for each lipid mixture were optimized on HEK293T cells for transfection and, using these values, the remaining cells were also transfected in absence (-FBS-FBS) and presence (-FBS+FBS) of serum. The transfection efficiency was higher with the CLs of shorter gemini spacers (n = 2, or 3). Each formulation expressed GFP on pDNA transfection and confocal fluorescence microscopy corroborated the results. C(16)C(2)C(16)/DOPE mixtures were the most efficient toward transfection among all the lipid mixtures and in presence of serum even better than the Lipofectamine2000, a commercial transfecting agent. Each lipid combination was safe and did not show any significant levels of toxicity. Probably, the presence of two coexisting lamellar structures in lipoplexes synergizes the transfection efficiency of the lipid mixtures which are plentiful in the lipoplexes formed by CLs with short spacer (n = 2, 3) than those with the long spacer (n = 5, 12).
Biomacromolecules 11/2012; · 5.48 Impact Factor
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ABSTRACT: Curcumin (Hcur) as a cellular imaging and PDT agent shows remarkable photocytotoxicity in HeLa cells in visible light of 400-700 nm giving IC(50) = 8.2 ± 0.2 μM and its degradation is arrested on formation of photocytotoxic dipyridophenazine (dppz) complex [VO(cur)(dppz)Cl] (IC(50) = 3.3 ± 0.4 μM), while both are less toxic in the dark.
Chemical Communications 06/2012; 48(62):7702-4. · 6.17 Impact Factor
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ABSTRACT: Iron(II) complexes [Fe(L)(2)](2+) as perchlorate (1-3) and chloride (1a-3a) salts, where L is 4'-phenyl-2,2':6',2″-terpyridine (phtpy in 1, 1a), 4'-(9-anthracenyl)-2,2':6',2″-terpyridine (antpy in 2, 2a) and 4'-(1-pyrenyl)-2,2':6',2″-terpyridine (pytpy in 3, 3a), were prepared and their photocytotoxicity studied. The diamagnetic complexes 1-3 having an FeN(6) core showed an Fe(III)-Fe(II) redox couple near 1.0V vs. saturated calomel electrode in MeCN-0.1M tetrabutylammonium perchlorate. Complexes 2 and 3, in addition, displayed a quasi-reversible ligand-based redox process near 0.0V. The redox and spectral properties are rationalized from the theoretical studies. The complexes bind to DNA in a partial intercalative mode. The pytpy complex efficiently photo-cleaves DNA in green light via superoxide and hydroxyl radical formation. The antpy and pytpy complexes exhibited a remarkable photocytotoxic effect in HeLa cancer cells (IC(50), ~9μM) in visible light (400-700nm), while remaining essentially nontoxic in dark (IC(50), ~90μM). Formation of reactive oxygen species (ROS) inside the HeLa cells was evidenced from the fluorescence enhancement of dichlorofluorescein upon treatment with the pytpy complex followed by photo-exposure. The antpy and pytpy complexes were used for cellular imaging. Confocal imaging and dual staining study using propidium iodide (PI) showed nuclear localization of the complexes.
Journal of inorganic biochemistry 06/2012; 116C:77-87. · 3.25 Impact Factor
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ABSTRACT: A red light for cancer cells: an iron(III) complex (1, see picture) that contains an anthracenyl fluorophore moiety and a catecholate ligand is a potent, metal-based PDT agent that efficiently photocleaves DNA in near-infrared light, has significant nuclear uptake, and high photocytotoxicity in red light by an apoptotic pathway in HeLa and MCF-7 cancer cells.
Angewandte Chemie International Edition 01/2012; 51(11):2658-61. · 13.45 Impact Factor
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ABSTRACT: A cationic amphiphile, cholest-5en-3β-oxyethyl pyridinium bromide (PY(+) -Chol), is able to efficiently disperse exfoliated graphene (GR) in water by the physical adsorption of PY(+) -Chol on the surface of GR to form stable, dark aqueous suspensions at room temperature. The GR-PY(+) -Chol suspension can then be used to solubilize Tamoxifen Citrate (TmC), a breast cancer drug, in water. The resulting TmC-GR-PY(+) -Chol is stable for a long time without any precipitation. Fluorescence emission and UV absorption spectra indicate the existence of noncovalent interactions between TmC, GR, and PY(+) -Chol in these suspensions. Electron microscopy shows the existence of segregated GR sheets and TmC 'ribbons' in the composite suspensions. Atomic force microscopy indicates the presence of 'extended' structures of GR-PY(+) -Chol, which grows wider in the presence of TmC. The slow time-dependent release of TmC is noticed in a reconstituted cell culture medium, a property useful as a drug carrier. TmC-GR-PY(+) -Chol selectively enhanced the cell death (apoptosis) of the transformed cancer cells compared to normal cells. This potency is found to be true for a wide range of transformed cancer cells viz. HeLa, A549, ras oncogene-transformed NIH3T3, HepG2, MDA-MB231, MCF-7, and HEK293T compared to the normal cell HEK293 in vitro. Confocal microscopy confirmed the high efficiency of TmC-GR-PY(+) -Chol in delivering the drug to the cells, compared to the suspensions devoid of GR.
Small 11/2011; 8(1):131-43. · 8.35 Impact Factor
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ABSTRACT: The most important objective of the present study was to explain why cationic lipid (CL)-mediated delivery of plasmid DNA (pDNA) is better than that of linear DNA in gene therapy, a question that, until now, has remained unanswered. Herein for the first time we experimentally show that for different types of CLs, pDNA, in contrast to linear DNA, is compacted with a large amount of its counterions, yielding a lower effective negative charge. This feature has been confirmed through a number of physicochemical and biochemical investigations. This is significant for both in vitro and in vivo transfection studies. For an effective DNA transfection, the lower the amount of the CL, the lower is the cytotoxicity. The study also points out that it is absolutely necessary to consider both effective charge ratios between CL and pDNA and effective pDNA charges, which can be determined from physicochemical experiments.
Journal of the American Chemical Society 11/2011; 133(45):18014-7. · 9.91 Impact Factor
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ABSTRACT: We explored the effect of a novel synthetic triterpenoid compound cyano enone of methyl boswellates (CEMB) on various prostate cancer and glioma cancer cell lines. CEMB displayed concentration-dependent cytotoxic activity with submicromolar lethal dose 50% (LD(50)) values in 10 of 10 tumor cell lines tested. CEMB-induced cytotoxicity is accompanied by activation of downstream effector caspases (caspases 3 and 7) and by upstream initiator caspases involved in both the extrinsic (caspase 8) and intrinsic (caspase 9) apoptotic pathways. By using short interfering RNAs (siRNA), we show evidence that knockdown of caspase 8, DR4, Apaf-1, and Bid impairs CEMB-induced cell death. Similar to other proapoptotic synthetic triterpenoid compounds, CEMB-induced apoptosis involved endoplasmic reticulum stress, as shown by partial rescue of tumor cells by siRNA-mediated knockdown of expression of genes involved in the unfolded protein response such as IRE1α, PERK, and ATF6. Altogether, our results suggest that CEMB stimulates several apoptotic pathways in cancer cells, suggesting that this compound should be evaluated further as a potential agent for cancer therapy.
Molecular Cancer Therapeutics 07/2011; 10(9):1635-43. · 5.23 Impact Factor
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ABSTRACT: Triterpenoids are pentacyclic secondary metabolites present in many terrestrial plants. Natural triterpenoids have been reported to exhibit anti-inflammatory and anti-carcinogenic activities. Here, we show that modifications of ring A of boswellic acid (2 cyano, 3 enone) resulted in a highly active growth inhibitory, anti-inflammatory, prodifferentiative and anti-tumour triterpenoid compound called cyano enone of methyl boswellates (CEMB). This compound showed cytotoxic activity on a number of cancer cell lines with IC₅₀ ranging from 0.2 to 0.6 μM. CEMB inhibits DNA synthesis and induces apoptosis in A549 cell line at 0.25 μM and 1 μM concentrations, respectively. CEMB induces adipogenic differentiation in 3T3-L1 cells at a concentration of 0.1 μM. Finally, administration of CEMB intra-tumourally significantly inhibited the growth of C6 glioma tumour xenograft in immuno-compromised mice. Collectively, these results suggest that CEMB is a very potent anti-tumour compound.
Journal of Biosciences 06/2011; 36(2):297-307. · 1.65 Impact Factor
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ABSTRACT: To understand the molecular pathogenesis of oral submucous fibrosis (OSF), which is a chronic inflammatory disease, gene expression profiling was performed in 10 OSF tissues against 8 pooled normal tissues using oligonucleotide arrays. Microarray results revealed differential expression of 5,288 genes (P ≤ 0.05 and fold change ≥ 1.5). Among these, 2,884 are upregulated and 2,404 are downregulated. Validation employing quantitative real-time PCR and immunohistochemistry confirmed upregulation of transforming growth factor-β1 (TGF-β1), TGFBIp, THBS1, SPP1, and TIG1 and downregulation of bone morphogenic protein 7 (BMP7) in OSF tissues. Furthermore, activation of TGF-β pathway was evident in OSF as demonstrated by pSMAD2 strong immunoreactivity. Treatment of keratinocytes and oral fibroblasts by TGF-β confirmed the regulation of few genes identified in microarray including upregulation of connective tissue growth factor, TGM2, THBS1, and downregulation of BMP7, which is a known negative modulator of fibrosis. Taken together, these data suggest activation of TGF-β signaling and suppression of BMP7 expression in the manifestation of OSF.
Growth factors (Chur, Switzerland) 05/2011; 29(4):119-27. · 2.47 Impact Factor
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ABSTRACT: We have synthesized five new cholesterol based gemini cationic lipids possessing hydroxyethyl (-CH(2)CH(2)OH) function on each head group, which differ in the length of the polymethylene spacer chain. These gemini lipids are important for gene delivery processes as they possess pre-optimized molecular features, e.g., cholesterol backbone, ether linkage and a variable spacer chain between both the headgroups of the gemini lipids. Cationic liposomes were prepared from each of these lipids individually and as a mixture of individual cationic gemini lipid and 1,2-dioleoyl phosphatidylethanolamine (DOPE). Each gemini lipid based formulation induced better transfection activity than that of their monomeric counterpart. One such gemini lipid with a -(CH(2))(12)- spacer, HG-12, showed dramatic increase in the mean fluorescence intensity due to the expression of green-fluorescence protein (GFP) in the presence of 10% FBS compared to the conditions where there was no serum. Other gemini lipids retained their gene transfection efficiency without any marked decrease in the presence of serum. The only exception was seen with the gemini with a -(CH(2))(3)- spacer, HG-3, which on gene transfection in the presence of 10% FBS lost ~70% of its transfection efficiency. Overall the gemini lipid with a -(CH(2))(5)- spacer, HG-5, showed the highest transfection activity at N/P (lipid/DNA) ratio of 0.5 and lipid : DOPE molar ratio of 2. Upon comparison of the relevant parameters, e.g., %-transfected cells, the amount of DNA transfected to each cell and %-cell viability all together against Lipofectamine 2000, one of the best commercial transfecting agents, the optimized lipid formulation based on DOPE/HG-5 was found to be comparable. In terms of its ability to induce gene-transfer in the presence of serum and shelf-life DOPE/HG-5 liposome was found to be superior to its commercial counterpart. Confocal imaging analysis confirmed that in the presence of 10% serum using a Lipid : DOPE of 1 : 4 and N/P charge ratio of 0.75 with 1.2 μg DNA per well, HG-5 is better than Lipofectamine 2000.
Organic & Biomolecular Chemistry 05/2011; 9(12):4600-13. · 3.70 Impact Factor
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ABSTRACT: Planar triazinium cationic species, from VO(2+)-assisted cyclization of 1-(2-thiazolylazo)-2-naphthol, shows efficient DNA intercalative binding, visible light-induced anaerobic plasmid DNA photocleavage activity and photocytotoxicity in HeLa and MCF-7 cancer cells by an apoptotic pathway with selective localization of the compound in the nucleus as evidenced from the nuclear staining and confocal imaging.
Chemical Communications 02/2011; 47(13):3954-6. · 6.17 Impact Factor
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Inorganica Chimica Acta 01/2011; · 1.85 Impact Factor
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Jayashree Ladha,
Sainitin Donakonda,
Shipra Agrawal,
Balaram Thota,
Mallavarapu R Srividya,
Sambandam Sridevi,
Arimappamagan Arivazhagan,
Kandavel Thennarasu,
Anandh Balasubramaniam,
Bangalore A Chandramouli,
Alangar Sattiyaranjandas Hegde, Paturu Kondaiah,
Kumaravel Somasundaram,
Vani Santosh,
Satyanarayana M R Rao
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ABSTRACT: Glioblastoma (GBM; grade IV astrocytoma) is a very aggressive form of brain cancer with a poor survival and few qualified predictive markers. This study integrates experimentally validated genes that showed specific upregulation in GBM along with their protein-protein interaction information. A system level analysis was used to construct GBM-specific network. Computation of topological parameters of networks showed scale-free pattern and hierarchical organization. From the large network involving 1,447 proteins, we synthesized subnetworks and annotated them with highly enriched biological processes. A careful dissection of the functional modules, important nodes, and their connections identified two novel intermediary molecules CSK21 and protein phosphatase 1 alpha (PP1A) connecting the two subnetworks CDC2-PTEN-TOP2A-CAV1-P53 and CDC2-CAV1-RB-P53-PTEN, respectively. Real-time quantitative reverse transcription-PCR analysis revealed CSK21 to be moderately upregulated and PP1A to be overexpressed by 20-fold in GBM tumor samples. Immunohistochemical staining revealed nuclear expression of PP1A only in GBM samples. Thus, CSK21 and PP1A, whose functions are intimately associated with cell cycle regulation, might play key role in gliomagenesis.
Cancer Research 08/2010; 70(16):6437-47. · 7.86 Impact Factor
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Vani Santosh,
Arimappamagan Arivazhagan,
Peddagangannagari Sreekanthreddy,
Harish Srinivasan,
Balaram Thota,
Mallavarapu R Srividya,
Marigowda Vrinda,
Sambandam Sridevi,
Bangalore C Shailaja,
Cini Samuel,
Krishnarao V Prasanna,
Kandavel Thennarasu,
Anandh Balasubramaniam,
Bangalore A Chandramouli,
Alangar S Hegde,
Kumaravel Somasundaram, Paturu Kondaiah,
Manchanahalli R S Rao
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ABSTRACT: Insulin-like growth factor (IGF)-binding protein (IGFBP) isoforms have been implicated in the pathogenesis of human neoplasms including glioma. In view of this, we evaluated the expression of IGFBP isoforms (IGFBP-2, -3, and -5) during malignant progression of astrocytoma and their prognostic significance in glioblastoma.
The expression of IGFBP isoforms was analyzed in diffusely infiltrating astrocytomas by real-time quantitative PCR (n = 203) and immunohistochemistry (n = 256). Statistical methods were used to assess their grade-specific expression pattern and mRNA-protein intercorrelation. Survival analyses were done on a uniformly treated, prospective cohort of adult patients with newly diagnosed glioblastoma (n = 136) by using Cox regression models.
The mean transcript levels of IGFBP-2 and -3 were significantly higher in glioblastomas (GBM) relative to anaplastic astrocytoma (AA), diffuse astrocytoma (DA), and controls whereas IGFBP-5 mRNA was higher in GBM relative to AA and controls (P < 0.05). By immunohistochemistry, the mean labeling index of all isoforms was significantly higher in GBM compared with AA, DA, and control (P < 0.05). A strong positive correlation was observed between their respective mRNA and protein expressions (P < 0.01). Multivariate analysis revealed IGFBP-3 expression (hazard ratio, 1.021; P = 0.030) and patient age (hazard ratio, 1.027; P = 0.007) to be associated with shorter survival in glioblastoma.
This study shows the associations of IGFBP-2, -3, and -5 expression with increasing grades of malignancy in astrocytomas. IGFBP-3 is identified as a novel prognostic glioblastoma biomarker. The strong correlation between their mRNA and protein expression patterns suggests their role in the pathogenesis of these tumors.
IGFBP isoforms have emerged as biomarkers with diagnostic and prognostic utility in astrocytomas.
Cancer Epidemiology Biomarkers & Prevention 06/2010; 19(6):1399-408. · 4.12 Impact Factor
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Peddagangannagari Sreekanthreddy,
Harish Srinivasan,
Durairaj Mohan Kumar,
Mamatha Bangalore Nijaguna,
Sambandam Sridevi,
Marigowda Vrinda,
Arimappamagan Arivazhagan,
Anandh Balasubramaniam,
Alangar Sathyaranjandas Hegde,
Bangalore A Chandramouli,
Vani Santosh,
Manchanahalli R S Rao, Paturu Kondaiah,
Kumaravel Somasundaram
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ABSTRACT: The aim of this study is to identify serum biomarkers with classification and prognosis utility for astrocytoma, in particular glioblastoma (GBM).
Our previous glioma microarray database was mined to identify genes that encode secreted or membrane-localized proteins. Subsequent analysis was done using significant analysis of microarrays, followed by reverse transcription-quantitative PCR (RT-qPCR) and immunohistochemical validation in tumor tissues, ELISA and Western blot validation in sera, and correlation with survival of GBM patients.
Significant analysis of microarrays identified 31 upregulated and 3 downregulated genes specifically in GBMs. RT-qPCR validation on an independent set of samples confirmed the GBM-specific differential expression of several genes, including three upregulated (CALU, CXCL9, and TIMP1) and two downregulated (GPX3 and TIMP3) novel genes. With respect to osteopontin (OPN), we show the GBM-specific upregulation by RT-qPCR and immunohistochemical staining of tumor tissues. Elevated serum OPN levels in GBM patients were also shown by ELISA and Western blot. GBM patients with high serum OPN levels had poorer survival than those with low serum OPN levels (median survival 9 versus 22 months respectively; P = 0.0001). Further, we also show high serum TIMP1 levels in GBM patients compared with grade II/III patients by ELISA and downregulation of serum GPX3 and TIMP3 proteins in GBMs compared with normal control by Western blot analysis.
Several novel potential serum biomarkers of GBM are identified and validated. High serum OPN level is found as a poor prognostic indicator in GBMs.
Identified serum biomarkers may have potential utility in astrocytoma classification and GBM prognosis.
Cancer Epidemiology Biomarkers & Prevention 06/2010; 19(6):1409-22. · 4.12 Impact Factor
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ABSTRACT: Limbal stem cell deficiency is a challenging clinical problem and the current treatment involves replenishing the depleted limbal stem cell (LSC) pool by either limbal tissue transplantation or use of cultivated limbal epithelial cells (LEC). Our experience of cultivating the LEC on denuded human amniotic membrane using a feeder cell free method, led to identification of mesenchymal cells of limbus (MC-L), which showed phenotypic resemblance to bone marrow derived mesenchymal stem cells (MSC-BM). To understand the transcriptional profile of these cells, microarray experiments were carried out.
RNA was isolated from cultured LEC, MC-L and MSC-BM and microarray experiments were carried out by using Agilent chip (4x44 k). The microarray data was validated by using Realtime and semiquntitative reverse transcription polymerase chain reaction.
The microarray analysis revealed specific gene signature of LEC and MC-L, and also their complementary role related to cytokine and growth factor profile, thus supporting the nurturing roles of the MC-L. We have also observed similar and differential gene expression between MC-L and MSC-BM.
This study represents the first extensive gene expression analysis of limbal explant culture derived epithelial and mesenchymal cells and as such reveals new insight into the biology, ontogeny, and in vivo function of these cells.
Molecular vision 01/2010; 16:1227-40. · 2.20 Impact Factor
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Anita B. Roberts,
Frédéric Rosa,
Nanette S. Roche,
John E. Coligan,
Mark Garfield,
Martha L. Rebbert, Paturu Kondaiah,
David Danielpour,
John H. Kehrl,
Sharon M. Wahl,
Igor B. Dawid,
Michael B. Sporn
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ABSTRACT: Abstract TGF-β2 and -β5 have been purified from medium conditioned by Xenopus cultured cells (XTC) and identified based on their N-terminal amino acid sequence analysis and biological activity. When applied in high concentrations, Xenopus TGF-β2, like porcine TGF-β2, induces expression of mesodermal markers from cultured Xenopus ectodermal explants, whereas TGF-β5 is inactive in this assay. However, the TGF-β's could be separated from the major mesoderm-inducing activity present in XTC medium. Xenopus TGF-β2 and -β5 are approximately equivalent to TGF-β1 in their abilities to inhibit the growth of mink lung CCL-64 cells, induce anchorage-independent growth of rat NRK cells, inhibit the proliferation and antibody secretion of human B-lymphocytes, and stimulate chemotaxis of human monocytes. These data establish the functional activity of TGF-β5 and suggest that more complex multicellular systems, in contrast to most isolated cells, discriminate between the different TGF-βs.
07/2009; 2(2):135-147.
