Generation of metastatic variants by transfection of a rat non-metastatic epithelial cell line with genomic DNA from rat prostatic carcinoma cells

Ke, Youqiang; Beesley, Carol; Smith, Paul; Barraclough, Roger; Rudland, Philip S.; Foster, Christopher S.

doi:10.1038/bjc.1998.45

Cited by 12 publications

(15 citation statements)

References 31 publications

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“…Our previous work, has found that the C-FABP gene was overexpressed in malignant breast and prostate epithelial cell lines. When C-FABP gene was transfected into the benign rat Rama 37 model cells (24,25) and the transfectants subsequently inoculated into Wistar Furth rats (Wistar OLA strain), a significant number of animals developed metastasis (11). Further studies demonstrated that C-FABP promoted tumourigenesis and metastasis of prostate cancer cells by facilitating angiogenesis through up-regulating VEGF gene (34,35).…”

Section: Discussionmentioning

confidence: 99%

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Expression of cutaneous fatty acid-binding protein (C-FABP) in prostate cancer: Potential prognostic marker and target for tumourigenicity-suppression

Morgan

Forootan²,

Adamson³

et al. 2008

Int J Oncol

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Abstract. C-FABP or E-FABP is a metastasis inducing gene over expressed in human prostate carcinomas. To study its prognostic significance, an archival set of prostate tissues was analysed immunohistochemically. Levels of both nuclear and cytoplasmic C-FABP expression in carcinoma cells were significantly higher than those in normal and BPH tissues and the increased C-FABP was significantly associated with a reduced patient survival time. To test the therapeutic potential of targeting C-FABP, a clone (Si-clone-2) of cells was established by interfering C-FABP expression in highly malignant PC-3M cells. Suppression of C-FABP in cancer cells significantly inhibited their proliferation and tumourigenicity in vitro. When Si-clone-2 cells were orthotopically implanted into the prostate gland of mouse, 2/13 mice produced primary tumours with an average size of 23±5 mg, and no metastasis was produced in any of the 13 animals. Whereas in the control group, all 14 mice produced primary tumours with an average size of 1450±370 mg and 9/14 (64.3%) produced metastasis. When inoculated subcutaneously, all 5 mice inoculated with control cells developed tumours from day 4, with an average size of 1471±544 mm 3 at 5 weeks after the inoculation; whereas Si-clone-2 cells produced no tumours in any of the 5 animals at any time-point, indicating the suppression occurred at the initiation stage. Our results suggest that C-FABP may be used as a potential prognostic marker to predict patient outcome and the increased C-FABP expression is a possible target to inhibit the malignant progression of prostate cancer cells.

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Section: Discussionmentioning

confidence: 99%

“…The SiRNA-C-FABP silencing construct was transfected into PC-3M cells by calcium phosphate co-precipitation technique as described previously (24,25). Exponentially growing PC-3M cells were seeded in 2 Petri dishes (9 cm in diameter) at a density of 0.5x10 6 /dish.…”

Section: Histological and Immunohistochemical Staining Of Tissuesmentioning

confidence: 99%

Expression of cutaneous fatty acid-binding protein (C-FABP) in prostate cancer: Potential prognostic marker and target for tumourigenicity-suppression

Morgan

Forootan²,

Adamson³

et al. 2008

Int J Oncol

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“…The remaining three cell lines in this group were the highly metastatic cell lines AT-3, MAT-Lu and AT-6.1, which can induce more than 75% of animals with metastasis. The last group of cell lines used in this study was the benign rat cell line Rama 37 (Dunnington et al, 1983) and its three malignant metastatic sublines established by transfecting the Rama 37 cells with DNA extracted from prostate carcinoma cells (Ke et al, 1998). The human and rat cell lines were grown as monolayer cultures in RPMI-1640 (human cells) and Dulbecco's modified Eagles medium (DMEM) (rat cells) media, supplemented with 10% (v/v) fetal calf serum, hydrocortisone (5 ng ml -1 ), penicillin (100 units ml -1 ) and streptomycin (100 mg ml -1 ), with either testosterone (5 ng ml -1 , for human and rat prostate cells), or insulin (5 ng ml -1 , for Rama 37 and its derivative lines).…”

Section: Cell Lines and Culture Conditionsmentioning

confidence: 99%

“…This glaring clinical fact has lent fresh impetus to the need for better understanding of mechanisms underlying metastatic behaviour. Progress on this front has been heightened recently by reproducible models of inducing metastases through transfection of fragmented DNA from malignant tumours into non-metastatic cells (Ke et al, 1998). Identification and characterization of genes involved in the metastatic phenotype of prostate cancer cells through these model systems hold the promise of propelling much needed advancements in therapy.…”

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confidence: 99%

“…Then, we have examined whether the ability of producing high level of VEGF can be transferred as the transformation of the metastatic capability of the cells by comparing the VEGF levels in three metastatic variants with that in their parental benign cells. These metastatic variants have been generated by transfection of a benign cell line with genomic DNA extracted from prostate carcinoma cells (Ke et al, 1998). Finally, we employed an in vivo angiogenesis assay to determine whether the VEGF produced by the malignant cells can induce new blood vessel formation.…”

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Chen¹,

Treweeke²,

Ke³

et al. 2000

Br J Cancer

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Vascular endothelial growth factor (VEGF) is one of the most potent factors for stimulating angiogenesis, an essential process required for expansion of primary tumour and dissemination of malignant cells. To investigate the possible role of VEGF in facilitating metastasis of prostate cancer via stimulating angiogenesis, we have used Northern and slot blotting, reverse transcription polymerase chain reaction, nucleotide sequence analysis and enzyme-linked immunosorbent assay to compare the VEGF expression in series of human and rat cell lines with either benign or malignant characteristics. We have also employed the chick chorioallantoic membrane (CAM) assay to measure the angiogenic activity of the VEGF derived from both benign and malignant cells. The level of VEGF mRNA expressed in the seven malignant human and rat cell lines is 3.5- to 10-fold higher than that expressed in the benign cell lines. The three metastatic variants, generated by transfection of a benign cell line with DNA extracted from prostate carcinoma cells, expressed 2.5 to 5 times more VEGF mRNA than their parental benign cells. While VEGF 121 and 165 were predominantly expressed by both the benign and malignant cells, the transcript representing VEGF 189 isoform was only detected in the malignant cells. At protein level, three human malignant cell lines produced more VEGF (2.7–7.9 ng ml −1 ) than the benign cell line (1.3 ng ml −1 ). CAM assay detected a VEGF-dependent angiogenic activity in the medium from malignant cells, but only a relatively weak VEGF-independent activity in the medium from benign cells. These results demonstrated that malignant cells did over-express VEGF and only the VEGF derived from malignant cells was angiogenically active. Thus, we suggest that the VEGF produced by malignant cells might play an important role in facilitating metastasis of prostatic cancer. © 2000 Cancer Research Campaign

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Relevance of a new rat model of osteoblastic metastases from prostate carcinoma for preclinical studies using zoledronic acid

Lamoureux

Ory

Battaglia

et al. 2007

Intl Journal of Cancer

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Animal models that mimic osteoblastic metastases associated with prostate carcinoma are required to improve the therapeutic options in humans. A new model was then developed and characterized in immunocompetent rats. The bisphosphonate zoledronic acid (ZOL) was tested to validate this model as a therapeutic application. Rat AT6-1 prostate tumor cells were characterized in vitro at the transcriptional (bone and epithelial markers) and functional (induction of mineralized nodules) levels. The bone lesions induced after their direct injection into the femur bone marrow were characterized by radiography, microscanner and histology analyses. ZOL effects were studied in vivo on bone lesion development and in vitro on AT6-1 cell proliferation, apoptosis and cell cycle analysis. Apart from epithelial markers, AT6-1 cells express an osteoblast phenotype as they express osteoblastic markers and are able to induce mineralized nodule formation in vitro. A disorganization of the trabecular bone at the growth zone level was observed in vivo after intraosseous AT6-1 cell injection as well as cortical erosion. The tumor itself is associated with bone formation as revealed by SEM analysis and polarized light microscopy. ZOL prevents the development of such osteoblastic lesions, related to a direct inhibitory effect on tumor cell proliferation independent of caspase 3 activation, but associated with cell cycle arrest. A new rat model of osteoblastic bone metastases was validated in immunocompetent rats and used to show the relevance of using ZOL in such lesions, as this compound shows bifunctional effects on both bone remodelling and tumor cell proliferation. ' 2007 Wiley-Liss, Inc.Key words: prostate carcinoma; osteoblastic metastases; experimental model; zoledronic acid Prostate adenocarcinoma, the second leading cause of cancer in men of North America is associated with significant morbidity and mortality due to skeletal metastases that commonly occur in advanced disease, affecting 80-100% of patients. 1,2 Therefore, new therapeutic strategies are needed to improve the prostate cancer patient's outcome. In contrast to bone metastases of breast cancer and myeloma, which are mainly osteolytic, a high percentage of prostate cancer metastases exhibit the radiographic appearance of osteoblastic lesions. 3 To investigate new targets for the development of novel therapies, experimental models that reproduce these osteoblastic bone lesions are required. However, there is a paucity of good preclinical models to study osteoblastic lesions.Animal models are important tools to investigate the pathogenesis and develop treatment strategies for bone metastases in humans. Because spontaneous bone metastasis in animals is uncommon, most animal models of bone metastasis must be derived experimentally. Experimental models of osteolytic, osteoblastic and mixed osteolytic/osteoblastic bone metastases include syngeneic transplantation of spontaneous occurring rodent cancers, chemical induction of cancers in selected strains of rats and mice, newly ...

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Generation of metastatic variants by transfection of a rat non-metastatic epithelial cell line with genomic DNA from rat prostatic carcinoma cells

Cited by 12 publications

References 31 publications

Expression of cutaneous fatty acid-binding protein (C-FABP) in prostate cancer: Potential prognostic marker and target for tumourigenicity-suppression

Expression of cutaneous fatty acid-binding protein (C-FABP) in prostate cancer: Potential prognostic marker and target for tumourigenicity-suppression

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Relevance of a new rat model of osteoblastic metastases from prostate carcinoma for preclinical studies using zoledronic acid

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