Gayatri Rath

Potent growth suppressive activity of curcumin in human breast cancer cells: Modulation of Wnt/β-catenin signaling

Abnormal activation of the Wnt/beta-catenin signaling pathway and subsequent upregulation of beta-catenin driven downstream targets-c-Myc and cyclin D1 is associated with development of breast cancer. The objective of our study was to determine if curcumin could modulate the key elements of Wnt pathway in breast cancer cells; an effect that might underscore its usefulness for chemoprevention/treatment of this malignancy. Curcumin showed a cytotoxic effect on MCF-7 cells with 50% inhibitory concentration (IC(50)) of 35microM; while IC(50) for MDA-MB-231 cells was 30microM. Treatment with low cytostatic dose of 20microM curcumin showed G(2)/M arrest in both breast cancer cells. The effect of curcumin (20microM) treatment on expression of Wnt/beta-catenin pathway components in breast cancer cells (MCF-7 and MDA-MB-231) was analyzed by immunofluorescence and Western blotting. Curcumin was found to effectively inhibit the expression of several Wnt/beta-catenin pathway components-disheveled, beta-catenin, cyclin D1 and slug in both MCF-7 and MDA-MB-231. Immunofluorescence analysis showed that curcumin markedly reduced the nuclear expression of disheveled and beta-catenin proteins. Further, the protein levels of the positively regulated beta-catenin targets-cyclin D1 and slug, were downregulated by curcumin treatment. The expression levels of two integral proteins of Wnt signaling, GSK3beta and E-cadherin were also altered by curcumin treatment. In conclusion, our data demonstrated that the efficacy of curcumin in inhibition of cell proliferation and induction of apoptosis might occur through modulation of beta-catenin pathway in human breast cancer cells.

Expression analysis of E-cadherin, Slug and GSK3β in invasive ductal carcinoma of breast

BackgroundCancer progression is linked to a partially dedifferentiated epithelial cell phenotype. The signaling pathways Wnt, Hedgehog, TGF-β and Notch have been implicated in experimental and developmental epithelial mesenchymal transition (EMT). Recent findings from our laboratory confirm that active Wnt/β-catenin signaling is critically involved in invasive ductal carcinomas (IDCs) of breast.MethodsIn the current study, we analyzed the expression patterns and relationships between the key Wnt/β-catenin signaling components- E-cadherin, Slug and GSK3β in IDCs of breast.ResultsOf the 98 IDCs analyzed, 53 (54%) showed loss/or reduced membranous staining of E-cadherin in tumor cells. Nuclear accumulation of Slug was observed in 33 (34%) IDCs examined. Loss or reduced level of cytoplasmic GSK3β expression was observed in 52/98 (53%) cases; while 34/98 (35%) tumors showed nuclear accumulation of GSK3β. Statistical analysis revealed associations of nuclear Slug expression with loss of membranous E-cadherin (p = 0.001); nuclear β-catenin (p = 0.001), and cytoplasmic β-catenin (p = 0.005), suggesting Slug mediated E-cadherin suppression via the activation of Wnt/β-catenin signaling pathway in IDCs. Our study also demonstrated significant correlation between GSK3β nuclear localization and tumor grade (p = 0.02), suggesting its association with tumor progression.ConclusionThe present study for the first time provided the clinical evidence in support of Wnt/β-catenin signaling upregulation in IDCs and key components of this pathway - E-cadherin, Slug and GSK3β with β-catenin in implementing EMT in these cells.

Soluble and membranous vascular endothelial growth factor receptor-1 in pregnancies complicated by pre-eclampsia

Vascular endothelial growth factor receptor-1 (VEGFR-1) is essential for the normal development and function of the placenta. Defective placental vasculogenesis and trophoblast function may lead to pre-eclampsia, a pregnancy-specific syndrome of hypertension and proteinuria. In order to study the association of VEGFR-1 with the development of pre-eclampsia, a cross-sectional study was carried out to evaluate the concentration of soluble VEGFR-1 (sVEGFR-1) in 360 serum samples and to analyze the expression of membranous VEGFR-1 in 40 placental samples of normal and pre-eclamptic pregnant women. Serum and placental samples at different gestational ages were collected from the Department of Obstetrics and Gynaecology, VMMC and Safdarjang Hospital, New Delhi. The serum levels of sVEGFR-1 and the expression of membranous VEGFR-1 were estimated by enzyme-linked immunosorbent assay and immunohistochemistry, respectively. The serum levels of sVEGFR-1 were seen to be positively increased (p=0.0001) in patients with pre-eclampsia at different gestational intervals as compared to the healthy pregnant women they were matched with. However, receiver operating characteristic (ROC) curve analysis showed a higher sensitivity (89.17%) and specificity (90.0%) in early onset (< or =34 weeks) in contrast with the late-onset (>34 weeks) pre-eclamptic group. Also, significant up-regulation of membranous VEGFR-1 immunoreactivity was observed in all placental cells (syncytiotrophoblast, cytotrophoblast, endothelial cells and Hofbauer cells) of pre-eclamptic groups in both < or =34 weeks (p=0.0001) and >34 weeks (p=0.0001) as compared to the normal group. Elevated sVEGFR-1 serum levels and up-regulated membranous VEGFR-1 expression in placenta denote abnormality in VEGF-mediated function in all placental cells, and thus may contribute to etiopathogenesis of pre-eclampsia. Nevertheless, this study also shows the possible diagnostic utility of sVEGFR-1 as a sensitive and specific biomarker for the early onset (< or =34 weeks) of pre-eclampsia.

Expression analysis of maspin in invasive ductal carcinoma of breast and modulation of its expression by curcumin in breast cancer cell lines

In breast cancer, maspin, a serine protease inhibitor, can suppress tumor growth and metastasis in vivo and tumor cell motility and invasion in vitro. The clinical significance of maspin expression in breast cancer, especially in the sequence of ductal carcinoma in situ (DCIS)-invasive cancer-lymph node metastasis is well known in the Western countries, but its status in the rapidly increasing breast cancers in India remains unknown. The present study was designed to determine the clinical significance of maspin expression in invasive ductal carcinomas of breast (IDCs) in North Indian population and modulation of its expression by curcumin. Immunohistochemical analysis of maspin showed loss or reduced cytoplasmic expression in 36 of 59 (61%) tumors. Furthermore, breast cancer cells (MCF-7 (wild type p53) and MDA-MB-231 (mutant p53)) were treated with curcumin and the effect on expression of maspin gene at transcription and translation levels was analyzed by RT-PCR, immunofluorescence and Western blotting. Maspin expression was also correlated with p53 and Bcl-2 levels. Curcumin inhibited cell growth, induced apoptosis and upregulated maspin gene expression in MCF-7 cells and these findings were further correlated with the upregulation of p53 protein and downregulation of Bcl-2, suggesting maspin mediated apoptosis in MCF-7 cells. To our knowledge this is the first report showing the upregulation of maspin expression by curcumin in breast cancer cells and taken together with the clinical data suggests a potential therapeutic role for curcumin in inducing maspin mediated inhibition of invasion of breast carcinoma cells.

Clinical significance of inactivated glycogen synthase kinase 3β in HPV-associated cervical cancer: Relationship with Wnt/β-catenin pathway activation.

To determine the role of inactivated GSK3β with respect to Wnt/β‐catenin pathway activation in HPV‐16/18‐associated cervical cancer.

Unusual Organization of Auriculotemporal Nerve and Its Clinical Implications

S J he auriculotemporal nerve is a highly significant tructure of infratemporal fossa. It may be comressed due to hypertrophy of the lateral pterygoid uscle leading to paresthesia over its area of distribuion and it may even be injured during temporomanibular joint surgery. Thus the precise anatomy of uriculotemporal nerve and its variations may prove eneficial to clinicians, especially to oral and maxilloacial surgeons. This study documents a rare variation f auriculotemporal nerve and its clinical implicaions.

HIF-1 Alpha and Placental Growth Factor in Pregnancies Complicated With Preeclampsia: A Qualitative and Quantitative Analysis

The pathophysiology of preeclampsia is not clearly understood worldwide. Hypoxia inducible factor 1α (HIF‐1α) is thought to be the preliminary factor for the hypoxic conditions prevailing in preeclampsia, which causes imbalance in the expression of angiogenic proteins. A proangiogenic protein, placental growth factor (PIGF), is reported to be dysregulated in preeclampsia. Therefore, this study focuses on the investigation of HIF‐1α and PIGF in preeclamptic conditions and a possible molecular association between them.

Apoptosis and Bcl-2 protein expression in human placenta over the course of normal pregnancy.

With 2 figures and 2 tables

Anomalous pectoral musculature

Anomalous disposition of pectoral muscles was encountered in an adult female cadaver on the left side. A prominent cleft separating the sternocostal and clavicular portions of the pectoralis major was noticed. The fibers of pectoralis major were partially fused with the deltoid, resulting in obliteration of the deltopectoral groove. Interestingly, cephalic vein was seen traversing superficial to the clavicular portion of the pectoralis major and pierced it to drain into the axillary vein. The pectoralis minor was inserted mainly on the coracoid process and few fibers were found blending with the coracobrachialis and short head of biceps brachii. Further, pectoralis minimus, a rare anatomic variant, was also observed lying superior to pectoralis minor. It was innervated by a twig from the lateral pectoral nerve at its superficial surface. Awareness of possibility of such anomalous muscles is important for surgeons operating on the chest wall.

Unilateral Double Axillary and Double Brachial Arteries: Embryological Basis and Clinical Implications

Este reporte describe la presencia de las arterias axilar y braquial duplicadas en el lado derecho de un cadaver adulto de sexo masculino. La arteria axilar derecha se bifurco en arterias axilares I y II, localizadas posteriormente al musculo pectoral menor. Ambas arterias, con sus respectivas ramas atravesaron bajo el musculo redondo mayor y continuaron como arterias braquiales I y II, respectivamente. La arteria axilar I, que se continuo como arteria braquial I, tuvo un trayecto superficial y tortuoso, mientras que la arteria axilar II fue de poco calibre y de localizacion profunda. Las arterias toracoacromial, toracica lateral y subescapular se originaron de la arteria axilar I. La arteria toracica superior se origino de la arteria toracoacromial. Las arterias circunflejas humerales anterior y posterior se originaron desde la arteria axilar II. La arteria braquial profundase origino de la arteria braquial II, mientras que a nivel de la linea intercondilea, la arteria ulnar se origino de la arteria braquial I. En la fosa cubital, la arteria braquial I se dividio en arterias radial e interosea comun, mientras que la arteria braquial II termino anastomosandose con la arteria braquial I. Las arterias ulnar, radial e interoseas comun continuaron como arterias independientes y tuvieron un trayecto y distribucion normales en el antebrazo. La persistencia de arterias axilar y braquial dobles tiene una base embriologica, pudiendo ser de interes para la embriologia experimental. El conocimiento de estas variaciones arteriales multiples, como el presente caso, es de importancia fundamental durante la medicion de flujo en los vasos sanguineos a traves de Doppler, para el diagnostico clinico y manejo quirurgico de enfermedades que afectan al miembro superior