For rapidly growing or proliferating cells like cancer cells, enhanced membrane synthesis is required
For rapidly growing or proliferating cells like cancer cells, enhanced membrane synthesis is required. observed that growth of melanoma cells cultured in conditioned medium (CM) from orlistat\treated adipocytes was reduced. Adipokines (leptin and resistin), via activating Akt and modulation of FASN as well as Cav\1 respectively, enhanced melanoma cell growth and proliferation. Together, we demonstrate that controlling body weight reduces adipose mass thereby diminishing melanoma progression. Therefore, strategic means of controlling obesity by reduced caloric diet or with antiobesity drugs treatment may render obesity\promoted tumor progression in check and prolong survival of patients. synthesis of fatty acids (Kridel et?al., 2004). Orlistat, at higher dosage, has been reported to exhibit antitumor properties as cancer cells rely on availability of fatty acids and related molecules for their survival (Menendez and Lupu, 2007; Seguin et?al., 2012). However, the equivalent anticancer dose of orlistat in humans, due to its severe adverse side effects, could be clinically unfeasible. Although a number of antiobesity drugs are available, diet\control interventions still remain to be the preferred line of therapy for effective management of obesity. Also, the role of dietary and nutritional factors towards cancer risk has been recently reported by many research groups (Kampman et?al., 2012; Prieto\Hontoria et?al., 2011; Rock et?al., 2012). However, the comprehensive investigations on the impact of effective management of obesity on tumor progression are lacking. Therefore, we hypothesized that controlling obesity would be an appropriate approach in minimizing the risk of obesity\promoted cancer progression. Hexaminolevulinate HCl In this study, we investigated the implications of therapeutic and dietary interventions for controlling obesity on the progression of melanoma. The underlying molecular events and role of specific adipokines were explored using appropriate and models. We demonstrate that controlling obesity is associated with normalization in levels of obesity\associated factors which parallels with reduction in melanoma progression and it may possibly be true for other cancer types too. 2.?Materials and methods 2.1. Experimental animals and diets Mice were procured from Experimental Animal Facility (EAF) at National Centre for Cell Science (NCCS), Pune, India. High fat diet (24% fat) was purchased from Provimi Animal Nutrition Pvt. Ltd., Bangalore, India, and normal diet (5% fat) was obtained from Amrut Hexaminolevulinate HCl Laboratory, Pune, India. Diet\induced obesity was developed in the mice by feeding with high fat diet as described previously (Pandey et?al., 2012). The composition Hexaminolevulinate HCl of the diets used is shown in Supplementary Table 1. Briefly, male C57BL/6J or female NOD/SCID mice (6C8 weeks old) were divided into normal diet (ND) and high fat diet (HFD) group. ND group (to all the mice. All animal experiments were carried out as per the requirement and guidelines of the Committee for the Purpose of Control and Supervision of Experiments on Animals (CPCSEA), Government of India, and after obtaining permission of the Institutional Animal Ethics Committee (IAEC). 2.2. Cells and Hexaminolevulinate HCl culture conditions Murine melanoma cells B16F10, human melanoma cells A375 and murine preadipocyte cells 3T3\L1 were procured from American Type Culture Collection (ATCC, Manassas, VA, USA) and maintained at our in\house cell repository at National Centre for Cell Science, Pune, India. Cells were routinely cultured in Dulbecco’s Modified Eagles Medium (DMEM) supplemented with 10% heat inactivated fetal bovine serum (Hyclone, UT, USA or Gibco, NY, USA), penicillin (100?U/ml) and streptomycin (100?g/ml) (Invitrogen Life Technologies, CA, USA) and maintained at 37?C in a 5% Rabbit polyclonal to ZNF19 CO2 humidified incubator (Thermo Fisher Scientific, OH, USA). 2.3. Serum biochemical analysis Blood glucose level was measured using rapid glucose analyzer (Accu\Chek Sensor Comfort, Roche Diagnostics, Mannheim, Germany) by collecting through an approved tail cap method. For serum collection, blood was collected by orbital sinus puncture and centrifuged at 6000 RPM at room temperature. Triglycerides (TG), cholesterol, LDLc and free fatty acids levels in fresh serum were estimated using colorimetric kits (Spinreact, Girona, Germany) as per the manufacturer’s instructions. Insulin,.