This apparent contradiction needs to be resolved before consideri

This apparent contradiction needs to be resolved before considering p8 as a target for treating can cer progression. ref 1 Conclusions In conclusion, we report in this paper that inhibition of p8 expression by an anti sense strategy increases the growth rate of both Panc 1 and BxPc 3 pan creatic cancer derived cells. Moreover, ERK and JNK mediated pathways down regulate p8 expression, whereas p38 Inhibitors,Modulators,Libraries and TGF 1 pathways induce p8 expression. Also, cell growth triggered by expression of a RAS mutated protein transcription is activated by TGF 1 through Smad4 in Interestingly, expression of p8 mRNA seems to be regu lated in a cell type and stimulus specific manner since, for example, p38 can induce p8 expression in response to or by 10% fetal calf serum induces down regulation of p8 expression.

Together, these results indicate that p8 is an intracellular cell growth inhibitor and that it is oppositely regulated by growth promoting or growth inhibiting fac tors in pancreatic cancer derived cells. Material and Methods Cell lines and cell culture conditions The human pancreatic cancer cell lines Panc 1 and BxPc 3 were Inhibitors,Modulators,Libraries a kind gift of Dr C. Susini and A. Hajri respectively. Panc 1 cells were grown in Dulbeccos modified Eagles medium supplemented with 10% fetal calf serum, 2 mM L glutamine, 100 IU/ml penicilin G and 100g/ml streptomycin. BxPc 3 were cultivated in RPMI 1640 medium in the presence of 2 mM L glutamine, 4. 5 g/L glucose, 10 mM Hepes, 1. 0 mM sodium pyruvate, 10% fetal calf serum and 100 IU/ml penicilin G and 100g/ml streptomycin.

Human recombinant TGF 1 was Inhibitors,Modulators,Libraries obtained from Sigma, and specific SB203580, U0126 and p8 expression in arrested and growing cells One million of Panc 1 or BxPc 3 cells were cultivated on 10 cm Petri plates in standard Inhibitors,Modulators,Libraries conditions. After 48 h, culture media were changed for fresh media with FCS restricted to 0. 1%, in order to Inhibitors,Modulators,Libraries stop growth. After 24 hours of growth arrest, culture medium was replaced either by medium with 10% fetal calf serum to resume cell growth or, as control, by medium with 0. 1% fetal calf serum. Twenty four hours later cells were recovered and RNA and protein extracted. p8 expression in TGF 1 treated Panc 1 cells One million of Panc 1 cells were cultivated in 10 cm cul ture dishes for 48 hours under standard conditions before TGF 1 treatment.

Human recombinant TGF 1 was added to cells, without changing the culture inhibitor licensed medium, and cells were collected 12 hours later for RNA and protein preparation. expressionrepresentationcancer derived cells regulating p8 SP600125 inhibitors were from Calbiochem and utilized at 10M. Expression plasmids Expression plamids encoding p38, Erk2, JNK, the wild type Raf and the Raf dominant negative were obtained from O Coso. Plamids encoding the constitutively activated type I TGF receptor, the dominant negative type II TGF receptor and the Smad4 dominant negative were obtained from R Urrutia and the wild type Smad4 was from C Heldin.

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