In Vivo Selection and Characterization of Metastatic Variants from Human Pancreatic Adenocarcinoma by Using Orthotopic Implantation in Nude Mice

Abstract

We determined whether the implantation of human pancreatic cancer cells into the pancreas of nude mice can be used to select variants with increasing metastatic potential. COLO 357 line fast-growing cells were injected into the spleen or pancreas of nude mice. Hepatic metastases were harvested, and tumor cells were reinjected into the spleen or pancreas. This cycle was repeated several times to yield cell lines L3.6sl (spleen to liver) and L3.6pl (pancreas to liver). The variant cells produced significantly higher incidence and number of lymph node and liver metastases than the parental cells. Their increased metastatic potential was associated with increased expression (mRNA and protein) of the proangiogenic molecules basic fibroblast growth factor, vascular endothelial growth factor, and interleukin-8. The metastatic cells also exhibited increased motility and invasiveness, which were associated with increased expression of collagenase type IV (MMP-9) and decreased expression of E-cadherin. Collectively, the data show that the orthotopic implantation of human pancreatic cancer cells in nude mice is a relevant model with which to study the biology of pancreatic cancer metastasis and to select variant cell lines with enhanced metastatic potential.

Effect of the vascular endothelial growth factor receptor‐2 antibody DC101 plus gemcitabine on growth, metastasis and angiogenesis of human pancreatic cancer growing orthotopically in nude mice

“…Orthotopic tumor cell injection into the tail of the pancreas was carried out as described previously. 15 …”

“…15,19 A positive reaction was visualized by incubating the slides with stable DAB (PCNA and Hypoxyprobe) or AEC (CD31) for 10 -20 min. The sections were rinsed with distilled water, counterstained with Gill hematoxylin solution for 1 min and mounted with Universal Mount (Research Genetics).…”

“…12,30 Consequently, a variety of different inhibitors of VEGF activity have been developed, including humanized neutralizing anti-VEGF monoclonal antibodies, 31 soluble VEGF-Rs, 32 anti-sense VEGF mRNA expressing constructs, 33 VEGF-toxin conjugates, 34 and inhibitors of VEGF-R function. 35 Because VEGF is implicated in the angiogenesis of human pancreatic cancer 15,19,36 and VEGF expression has been shown to be associated with a poor prognosis in human pancreatic cancer, [37][38][39] we investigated the efficacy of a VEGF-R2 antibody, DC101, with or without gemcitabine on primary tumor growth and metastasis of human pancreatic cancers growing orthotopically in nude mice. Treatment with gemcitabine or DC101 alone resulted in an ϳ65% and ϳ70% reduction of primary pancreatic tumor volume, respectively, as compared to control.…”

CD44v6‐Peptide Functionalized Nanoparticles Selectively Bind to Metastatic Cancer Cells

“…The cells were injected into the pancreas of NCI‐nu as previously described 24. Mice were injected i.p.…”

Potential of tumour cells for delivering oncolytic viruses

“…11 Furthermore, it is possible to select metastasis-prone populations from the primary tumour or its draining lymph nodes or from established metastases with the help of tissue-specific endothelial matrices, hypoxic growth conditions or distinct cell markers such as CD44. [12][13][14][15] Carriers generated ex tempore from these ATCs should be endowed with an enhanced capacity to pick up the trail of neoplastic dissemination and to transport their OV load to the right location after systemic application. This was recently demonstrated by our finding that rat hepatoma cells can transfer H-1PV to lung metastases originating from these cells and can initiate oncolysis within these metastases.…”