Molecular alterations of tumor-associated markers in human pancreatic tumor cell lines (based on [27], [28]).

a<p>mutated;</p>b<p>deleted,</p>c<p>wild type,</p>*<p>no consistent information available.</p

Cytochrome 2B1 protein expression in PCCWmCMV-transduced and non-transduced parental cell lines.

<p>Total cellular lysates from 8×10⁵ cells were separated on a 10% polyacrylamide gel under denaturing conditions. After blotting, CYP2B1 protein was detected with a CYP-specific antibody.</p

Summary of orthotopical tumor growth.

<p>Number of tumors growing after i.p. injection into mice and percentage of tumors that infiltrated the pancreatic tissue.</p>a<p>infiltrative tumor growth rate into the tail of the pancreas; percentages indicate the proportion of mice with infiltrative tumor growth among those with orthotopic tumor growth.</p

Tumor growth development of subcutaneously growing tumors.

<p>5×10⁶ cells of each pancreatic cell line were subcutaneously injected into the left flank of C.B-17/IcrHsd-<i>Prkcd^scid Lyst^bg</i> mice at day 0. Visible tumors were measured twice a week with a caliper and size was calculated by the formula “length×width×width/2”.</p

Cytosine deaminase gene expression in PCCDWmCMVpuro-transduced and in parental cell lines.

<p>Total cellular RNA was isolated from indicated cell lines and mRNA was reverse transcribed using oligo(dT) primers. CD- as well as GAPH-specific gene fragments were amplified using specific primers and separated on an agarose gel showing the expected sizes of ∼480 bp for CD and ∼350 bp for GAPDH. The cDNA amount used for PCR amplification was adjusted to yield comparable amounts of the GAPDH gene fragment.</p

Pathohistological analyses of tumor tissues.

<p>(<b>2A</b>) Sections of pancreatic tumors derived of the indicated pancreatic cell lines were stained with haematoxylin and eosin. The Hs-766T specimen was dissected from a subcutaneous tumor; all other samples were taken from orthotopically grown tumors. (<b>2B</b>) Different sections of a BxPC-3-derived tumor were subjected to immunohistological analysis with antibodies against Mucin-1 (MUC-1), pan-cytokeratin (Pan-CK), and α-smooth muscle actin (SMA). The black bar represents a length of 53 µm, except for anti-MUC-1 staining (27 µm).</p

Sensitivity of human pancreatic cancer cell lines towards ifosfamide.

<p>Cells stably transduced with the CYP2B1 gene as well as non-transduced parental cells (mock) were cultured in increasing concentrations of ifosfamide for five days as described in Materials and Methods. LD₅₀ values were calculated using Excel Fit software. Each experiment was performed in quadruplicates. Data from three independent experiments are shown.</p

Sensitivity of human pancreatic cancer cell lines towards 5-FC and 5-FU.

<p>Cells stably transduced with the yeast CD gene as well as non-transduced parental cells (mock) were cultured in increasing concentrations of 5-FC or 5-FU for five days as described in Materials and Methods. LD₅₀ values were calculated using Excel Fit software. Each experiment was performed in quadruplicates. Shown are data from three independent experiments.</p