RNAi experiment The RNAi experiment was performed with the Lipofectamine RNAiMAX reagent Enzastaurin PKC in accordance with the manufacturers instructions. The sequences of siRNA for c Myc were 5 AGA CCU UCA UCA AAA ACA UTT 3 and 5 AUG UUU UUG AUG AAG GUC UCG 3, which were designed by Ambion, and the non silencing control siRNA was Inhibitors,Modulators,Libraries purchased from Invitrogen. After incubation with the siRNA for 48 h at 37 C, the mRNA expressions of c Myc and Angptl4 were quantitatively determined by real time PCR. Short hairpin RNA targeting the Angptl4 including entry vector was designed and prepared by Invitrogen. The shRNA was subcloned to a retrovirus vector and Inhibitors,Modulators,Libraries used in the experiments as described in a previous study. ChIP assay The ChIP assay was performed using the ChIP IT Express kit, in accordance with the manufacturers instructions.

LN229 cells were fixed with 1% formaldehyde for 10 min. The cells were then washed, lysed, and sonicated to reduce DNA lengths to the range of 200 to 1500 bp. The chromatin/DNA complexes were incubated with antibodies to c Myc or IgG overnight at 4 C. The immune complexes were precipitated, eluted, reverse crosslinked, and treated with proteinase K. After extraction of the DNA fragments, Inhibitors,Modulators,Libraries real time PCR analysis was performed using Power SYBR green PCR master mixes. The primer for the promoter of Angptl4 was purchased from BioScience The predicted PCR product in cluded a c Myc binding sequence. Relative enrichment was comparatively calculated using IgG negative control as de scribed in eBioScience instructions.

Statistical analysis Significant differences were analyzed by an unpaired Students t test or analysis of variance with Tukeys post hoc test using the GraphPad Prism software. p 0. 05 was considered to indicate statisti cally significant difference. Background Melanoma is the most aggressive form of skin Inhibitors,Modulators,Libraries cancer. Its incidence and mortality have risen dramatically in all de veloped countries during the last half century. Although most cases of melanoma are diagnosed early and surgically resected, later stages of this tumour have very poor survival rates because of the lack of available effective therapies. Recently, promising therapeutic approaches for mel anoma management have been introduced into the clinical practice, based mostly on the use of small molecule inhibi tors directed against oncogenic molecular targets as well as on immunotherapy.

However, a high molecular het erogeneity of melanoma tumours and a complex network 1 of proliferation and survival Inhibitors,Modulators,Libraries pathways involved in its pathogenesis have been reported. For this reason, there is a growing interest in seeking pharmacological agents that could target multiple gene products in order to interfere, at different levels, with pathogenetic pathways in melanoma. During the last decades, several dietary agents have been reported to exert anticancer inhibitor Seliciclib activity.