Characterization of novel carcinoma cell lines for the analysis of therapeutical strategies fighting pancreatic cancer

Abstract

BackgroundPreclinical evaluations of chemotherapies depend on clinically relevant animal models for pancreatic cancer. The injection of syngeneic murine adenocarcinoma cells is one efficient option to generate carcinomas in mice with an intact immune system. However, this option is constrained by the paucity of appropriate cell lines.ResultsThe murine pancreatic adenocarcinoma cell lines 6606PDA and 7265PDA were compared to the 6606l cell line isolated from a liver metastasis from mice suffering from pancreatic cancer. In tissue culture 6606PDA and 6606l proliferated faster than 7265PDA. 7265PDA cells were, however, significantly more sensitive to gemcitabine as assessed by BrdU-incorporation and trypan blue exclusion assays in vitro. Within 1 week after injection of either one of these three cell lines into the pancreas of C57BL/6J mice, carcinomas were observed by T2 weighted magnetic resonance imaging and histology. Three weeks after injecting 6606PDA or 6606l cells large carcinomas could be characterized, which were surrounded by extensive desmoplastic reaction. After injection of 7265PDA cells, however, remission of cancer was observed between the first and the third week. Compared to 6606PDA cell derived carcinomas a higher apparent diffusion coefficient was quantified by diffusion weighted magnetic resonance imaging in these tumors. This correlated with reduced cancer cell density observed on histological sections.ConclusionAll three cell lines can be used in vitro for testing combinatorial therapies with gemcitabine. The 6606PDA and 6606l cell lines but not the 7265PDA cell line can be used for evaluating distinct therapies in a syngeneic carcinoma model using C57BL/6J mice. Diffusion-weighted MRI proved to be an appropriate method to predict tumor remission.

Intratumoral heterogeneity of the therapeutical response to gemcitabine and metformin

“…Treating mice with both substances lead to an even larger reduction of the tumor weight (Figure 2A ). Independent of treatment, all tumors had similar histological features as described previously [ 19 , 20 ]. They were cyst-like, completely encapsulated by fibroblast like cells and had few necrotic areas close to the encapsulation (0–120 μm distance from the desmoplastic reaction), but more necrotic areas at 360–570 μm distance from the desmoplastic reaction (Figure 2B and 2C ).…”

“…The murine cell line, 6606PDA and 7265PDA were a gift of Prof. Tuveson (University of Cambridge, UK) and were grown in DMEM high glucose medium (Biochrom GmbH, Berlin, Germany) as previously described [ 19 , 20 ]. The human MIA PaCa-2 cells were ordered from ATCC (LGC Standards GmbH, Wesel, Germany).…”

“…Male C57BL/6J mice were purchased from The Jackson Laboratory (Bar Harbor, ME) and bred in our local animal facility. As published previously, laparotomy was performed on anesthetized mice (1.2–2.5% isoflurane), 2.5 × 10 5 carcinoma cells were injected into the pancreas head, and the abdominal cavity was closed by sutures [ 20 ]. For pain relief 5 mg/kg carprofen (Pfizer GmbH, Berlin, Germany) was injected (sc) before surgery and 800 mg/L metamizol (Ratiopharm GmbH, Ulm, Germany) was added to the drinking water until euthanasia of the mice.…”

“…Data presentation and statistics were performed as described previously [ 19 , 20 ]. Box plots indicate the median, the 25th and 75th percentiles in the form of a box, and the 5th and 95th percentiles as whiskers.…”

Investigation of Anti-Tumor Effects of an MLK1 Inhibitor in Prostate and Pancreatic Cancers

“…The mouse pancreatic cell lines (6606PDA, 6606I, and 7265PDA) were kindly provided by Dr. David Tuveson (Cambridge University) via a material transfer agreement. The three pancreatic cancer cell lines were maintained in Dulbecco’s modified Eagle medium/F12 (Invitrogen, Carlsbad, CA, USA) with 10% fetal calf serum, according to established procedures [ 10 ]. In experiments with an AR inhibitor, MDV3100 (Selleckchem, Houston, TX, USA), cells were incubated with 10 µM MDV3100 for 24 h in 10% serum-containing RPMI 1640 medium.…”