Ann Surg Treat Res.  2017 Oct;93(4):173-180. 10.4174/astr.2017.93.4.173.

Establishing a colorectal cancer liver metastasis patient-derived tumor xenograft model for the evaluation of personalized chemotherapy

• Jung J
• Kim J
• Lim HK
• Kim KM
• Lee YS
• Park JS
• Yoon DS

• KMID: 2392366
• DOI: http://doi.org/10.4174/astr.2017.93.4.173

Abstract

PURPOSE
In order to suggest optimal anticancer drugs for patient-tailored chemotherapy, we developed a colorectal cancer (CRC)-liver metastasis patient-derived tumor xenograft (PDTX) model.
METHODS
Tissue obtained from a patient with CRC-liver metastasis (F0) was transplanted in a nonobese female mouse with diabetic/severe combined immune deficiency (F1) and the tumor tissue was retransplanted into nude mice (F2). When tumor volumes reached ~500 mm³, the F2 mice were randomly divided into 4 groups (n = 4/group) of doxorubicin, cisplatin, docetaxel, and nontreated control groups. The tumor tissues were investigated using H&E staining, terminal deoxynucleotidyl transferase dUTP nick end labeling assays, and immunohistochemistry. To determine where the mutant allele frequencies varied across the different passages, we isolated genomic DNA from the primary tumor, liver metastasis, and PDTX models (F1/F2).
RESULTS
The physiological properties of the tumor were in accord with those of the patient's tumors. Anticancer drugs delayed tumor growth, inhibited proliferation, and caused apoptosis. Histological assessments revealed no observable heterogeneity among the intragenerational PDTX models. Target exon sequencing analysis without high-quality filter conditions revealed some genetic variations in the 83 cancer-related genes across the generations. However, when de novo mutations were defined as a total count of zero in F0 and ≥5 in F2, exactly prognostic impact of clone cancer profiling (EGFR, KRAS, BRAF, PIK3CA, NRAS, APC and TP53) were detected in the paired.
CONCLUSION
A CRC liver metastasis PDTX model was established for the evaluation of chemotherapeutic efficacy. This model retained the physiological characters of the patient tumors and potentially provides a powerful means of assessing chemotherapeutic efficacy.

Keyword

Colorectal neoplasms; Metastasis; Xenograft model antitumor assays; Sequence analysis

MeSH Terms

Animals
Apoptosis
Cisplatin
Clone Cells
Colorectal Neoplasms*
DNA
DNA Nucleotidylexotransferase
Doxorubicin
Drug Therapy*
Exons
Family Characteristics
Female
Gene Frequency
Genetic Variation
Heterografts*
Humans
Immunohistochemistry
Liver*
Mice
Mice, Nude
Neoplasm Metastasis*
Population Characteristics
Sequence Analysis
Xenograft Model Antitumor Assays
Cisplatin
DNA
DNA Nucleotidylexotransferase
Doxorubicin

Figure

• 

  Fig. 1 Establishment of a patient-derived colorectal cancer-liver metastasis xenograft model. (A) Schematic of the patientderived tumor xenograft (PDTX) model. (B) Tumor growth PDTX model (F2). Tumor tissues were retransplanted into Balb/c nu/nu mice (n = 18). Results are presented as mean ± standard deviation.

• 

  Fig. 2 Histologic evaluation of a patient-derived colorectal cancer-liver metastasis xenograft model. (A) Histology of a patient tumor and of the patient-derived tumor xenograft F1- and F2-derived tumors. Tissue sections were stained with H&E (×400). (B) Identification of the metastasis model. Tissue sections were immunostained for CK7 and CK20 (×200).

• 

  Fig. 3 Chemotherapeutic effects of anticancer drugs in the patient-derived tumor xenograft model. (A) Tumor growth delay of anticancer drugs. (B) Mouse body weights. Results are presented as mean ± standard deviation (n = 4). CDDP, cisplatin; DOX, doxorubicin; DTX, docetaxel.

• 

  Fig. 4 Characterization of tumor tissues after treatment with anticancer drugs. (A) Immunostaining with Ki-67 as a marker of proliferation. Tissue sections were stained with hematoxylin (×200). (B) Tumor tissue apoptosis was quantified using the TUNEL assay. Brown dots represent apoptotic cells. Tissues were observed under a microscope (×400). (C) Tumor necrosis. Tissue sections were stained with H&E (×200).

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