LNCaP Xenograft Model

LNCAP Xenograft Altogen Labs

LNCaP xenograft model

Prostate cancer is the most common malignancy and the third primary cause of cancer deaths in males in the United States, as per the American Cancer Society. Xenograft preclinical studies aid in the discovery of new treatment regimens for prostate cancer patients. The LNCaP epithelial cell line was isolated from the supraclavicular lymph node metastasis of a Caucasian male in 1977. LNCaP can grow in aggregates or as a single cell. The LNCaP cell line has a high affinity for androgen receptors and is receptive to hormones. A 2015 study by Yang et al. published in PLoS One, investigated if the combination of quercetin and 2-Methoxyestradiol (2-ME) could inhibit the LNCaP xenograft tumor growth in vivo. According to the article, the combination of quercetin and 2-ME blocks tumor growth in the LNCaP xenograft model and decreases side effects of quercetin or 2-ME alone. Therefore, it could be an innovative treatment approach in human prostate cancer. The LNCap cell line (human prostate) is used to create the CDX (Cell Line Derived Xenograft) LNCaP xenograft mouse model. The LNCaP xenograft model is androgen sensitive (mutated androgen receptor, AR T877A) and can be treated with modalities such as an androgen receptor antisense oligonucleotide or bicalutamide.

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Basic study design

  1. Cells are grown under asceptic conditions prior to collection and at exponential growth.
  2. LNCaP cells are then collected and viability is determined.  Trypan blue is utilized to ensure a minimum of 98% viability of the cells.
  3. Cell suspension concentrations are adjusted such that in 100 uL of the matrigel + LNCaP cell suspension there are one million cells (volume of injection is 100 uL).  Inoculations are made into  the s.c. flank of a hind leg per mouse.  The mice are nu/nu and 10 weeks old.
  4. Calipers are applied for monitoring tumor growth, with tumor sizes of 50-150 mm3 needed for study initiation.  Animals are then sorted into necessary groupings and client provided compounds are administered.
  5. Tumor measurements and mouse weights are recorded until end of study.  Tumors are removed, their weights recorded and documented digitally.
  6. Tissue samples can be isolated for nucleic acids, submersed in RNAlater, prepared for histological analysis or snap frozen.

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LNCaP Xenograft Model

Xenograft animal models are used to assess the effectiveness of drugs against specific types of cancer. New medicines are tested on staged tumor growths that have been engrafted via subcutaneous or orthotopic inoculation in an immunocompromised mouse or rat model. All clinically approved anti-cancer agents have been evaluated with conventional preclinical in vivo models. Xenograft studies can be highly complex, starting with the selection of the appropriate animal model, choice of tumorigenic cell line, administration method, dosing, analysis of tumor growth rates and tumor analysis (histology, mRNA and protein expression levels).

Altogen Labs provides an array of laboratory services using over 30 standard Cell Line Derived Xenograft (CDX) models and over 20 PDX models. Researchers investigating the role of specific proteins or gene products in regulating tumor growth can benefit from development of protein overexpression (genetically engineered to ectopically express proteins, tumor suppressors, or oncogenes) and RNAi cell lines with long term gene silencing. Altogen Labs provides quantitative gene expression analysis of mRNA expression (RT-PCR) and protein expression analysis using the WES system (ProteinSimple).

The dosing of the experimental compound of interest is initiated, for a staged study, when the mean tumor size reaches a specified volume (typically 50-100 mm3). In an unstaged study, the dosing of the compound of interest is initiated immediately after xenografting. Mice are dosed once or twice a day for 28 days (or other desired study duration) via the chosen route of administration. Tumor volume (mm3) is calculated via the “(W x W x L) / 2” formula, where W is tumor width and L is tumor length.

Animal handling and maintenance at the Altogen Labs facility is IACUC-regulated and GLP-compliant. Following acclimatization to the vivarium environment, mice are sorted according to body mass. The animals are examined daily for tumor appearance and clinical signs. We provide detailed experimental procedures, health reports and data (all-inclusive report is provided to the client that includes methods, results, discussion and raw data along with statistical analysis). Additional services available include collection of tissue, histology, isolation of total protein or RNA and analysis of gene expression. Our animal facilities have the flexibility to use specialized food or water systems for inducible gene expression systems.

Following options are available for the LNCaP xenograft model:

  • LNCaP Tumor Growth Delay (TGD; latency)
  • LNCaP Tumor Growth Inhibition (TGI)
  • Dosing frequency and duration of dose administration
  • Dosing route (intravenous, intratracheal, continuous infusion, intraperitoneal, intratumoral, oral gavage, topical, intramuscular, subcutaneous, intranasal, using cutting-edge micro-injection techniques and pump-controlled IV injection)
  • LNCaP tumor immunohistochemistry
  • Alternative cell engraftment sites (orthotopic transplantation, tail vein injection and left ventricular injection for metastasis studies, injection into the mammary fat pad, intraperitoneal injection)
  • Blood chemistry analysis
  • Toxicity and survival (optional: performing a broad health observation program)
  • Gross necropsies and histopathology
  • Positive control group employing cyclophosphamide, at a dosage of 50 mg/kg administered by intramuscular injection to the control group daily for the study duration
  • Lipid distribution and metabolic assays
  • Imaging studies: Fluorescence-based whole body imaging, MRI

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