Limits...
Functional Characterization and Drug Response of Freshly Established Patient-Derived Tumor Models with CpG Island Methylator Phenotype.

Maletzki C, Huehns M, Knapp P, Waukosin N, Klar E, Prall F, Linnebacher M - PLoS ONE (2015)

Bottom Line: Initial DNA fingerprint analysis confirmed identity with the patient in all four cases.These freshly established cells showed characteristic features associated with the CIMP-phenotype (HROC40: APCwt, TP53 mut, KRAS mut; 3/8 marker methylated; HROC43: APC mut, TP53 mut, KRAS mut; 4/8 marker methylated; HROC60: APCwt, TP53 mut, KRASwt; 4/8 marker methylated; HROC183: APC mut, TP53 mut, KRAS mut; 6/8 marker methylated).Of note, most cell lines were sensitive towards "non-classical" CRC standard drugs (sensitivity: Gemcitabin > Rapamycin > Nilotinib).

View Article: PubMed Central - PubMed

Affiliation: Molecular Oncology and Immunotherapy, University of Rostock, Rostock, Germany.

ABSTRACT
Patient-individual tumor models constitute a powerful platform for basic and translational analyses both in vitro and in vivo. However, due to the labor-intensive and highly time-consuming process, only few well-characterized patient-derived cell lines and/or corresponding xenografts exist. In this study, we describe successful generation and functional analysis of novel tumor models from patients with sporadic primary colorectal carcinomas (CRC) showing CpG island methylator phenotype (CIMP). Initial DNA fingerprint analysis confirmed identity with the patient in all four cases. These freshly established cells showed characteristic features associated with the CIMP-phenotype (HROC40: APCwt, TP53 mut, KRAS mut; 3/8 marker methylated; HROC43: APC mut, TP53 mut, KRAS mut; 4/8 marker methylated; HROC60: APCwt, TP53 mut, KRASwt; 4/8 marker methylated; HROC183: APC mut, TP53 mut, KRAS mut; 6/8 marker methylated). Cell lines were of epithelial origin (EpCAM+) with distinct morphology and growth kinetics. Response to chemotherapeutics was quite individual between cells, with stage I-derived cell line HROC60 being most susceptible towards standard clinically approved chemotherapeutics (e.g. 5-FU, Irinotecan). Of note, most cell lines were sensitive towards "non-classical" CRC standard drugs (sensitivity: Gemcitabin > Rapamycin > Nilotinib). This comprehensive analysis of tumor biology, genetic alterations and assessment of chemosensitivity towards a broad range of (chemo-) therapeutics helps bringing forward the concept of personalized tumor therapy.

Show MeSH

Related in: MedlinePlus

Response to Aza-based drug combinations.Representative quantitative analysis of HROC40 (left graph) and HROC183 cells (right graph) treated with Aza, standard drugs or combinations thereof. Cells received two (upper panel) or four (lower panel) treatment cycles in concentrations as indicated in the material and methods section. Cytotoxicity was quantified upon crystal violet staining and measurement at 570 nm (reference wavelength: 620 nm). Results show the mean + standard deviation of three independent experiments.
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4664421&req=5

pone.0143194.g006: Response to Aza-based drug combinations.Representative quantitative analysis of HROC40 (left graph) and HROC183 cells (right graph) treated with Aza, standard drugs or combinations thereof. Cells received two (upper panel) or four (lower panel) treatment cycles in concentrations as indicated in the material and methods section. Cytotoxicity was quantified upon crystal violet staining and measurement at 570 nm (reference wavelength: 620 nm). Results show the mean + standard deviation of three independent experiments.

Mentions: Additionally, Azacytidine alone and in combination with chemotherapeutic drugs was studied. Here, IC50 values for Azacytidine were above 30 μM, even in the CIMP-H cell line HROC183. Longer incubation time increased the antitumoral effect of Azacytidine (Fig 6, lower panel). Best effects were seen in HROC183 cells. Drug combinations, either given simultaneous or metronomic, slightly increased the antitumoral effect of Azacytidine (representative data are given in Fig 6). However, the different drug combinations were not better than single drug treatments and in virtually all cases; even antagonistic effects were observed (e.g. Azacytidine + 5-FU or Irinotecan; Fig 6).


Functional Characterization and Drug Response of Freshly Established Patient-Derived Tumor Models with CpG Island Methylator Phenotype.

Maletzki C, Huehns M, Knapp P, Waukosin N, Klar E, Prall F, Linnebacher M - PLoS ONE (2015)

Response to Aza-based drug combinations.Representative quantitative analysis of HROC40 (left graph) and HROC183 cells (right graph) treated with Aza, standard drugs or combinations thereof. Cells received two (upper panel) or four (lower panel) treatment cycles in concentrations as indicated in the material and methods section. Cytotoxicity was quantified upon crystal violet staining and measurement at 570 nm (reference wavelength: 620 nm). Results show the mean + standard deviation of three independent experiments.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4664421&req=5

pone.0143194.g006: Response to Aza-based drug combinations.Representative quantitative analysis of HROC40 (left graph) and HROC183 cells (right graph) treated with Aza, standard drugs or combinations thereof. Cells received two (upper panel) or four (lower panel) treatment cycles in concentrations as indicated in the material and methods section. Cytotoxicity was quantified upon crystal violet staining and measurement at 570 nm (reference wavelength: 620 nm). Results show the mean + standard deviation of three independent experiments.
Mentions: Additionally, Azacytidine alone and in combination with chemotherapeutic drugs was studied. Here, IC50 values for Azacytidine were above 30 μM, even in the CIMP-H cell line HROC183. Longer incubation time increased the antitumoral effect of Azacytidine (Fig 6, lower panel). Best effects were seen in HROC183 cells. Drug combinations, either given simultaneous or metronomic, slightly increased the antitumoral effect of Azacytidine (representative data are given in Fig 6). However, the different drug combinations were not better than single drug treatments and in virtually all cases; even antagonistic effects were observed (e.g. Azacytidine + 5-FU or Irinotecan; Fig 6).

Bottom Line: Initial DNA fingerprint analysis confirmed identity with the patient in all four cases.These freshly established cells showed characteristic features associated with the CIMP-phenotype (HROC40: APCwt, TP53 mut, KRAS mut; 3/8 marker methylated; HROC43: APC mut, TP53 mut, KRAS mut; 4/8 marker methylated; HROC60: APCwt, TP53 mut, KRASwt; 4/8 marker methylated; HROC183: APC mut, TP53 mut, KRAS mut; 6/8 marker methylated).Of note, most cell lines were sensitive towards "non-classical" CRC standard drugs (sensitivity: Gemcitabin > Rapamycin > Nilotinib).

View Article: PubMed Central - PubMed

Affiliation: Molecular Oncology and Immunotherapy, University of Rostock, Rostock, Germany.

ABSTRACT
Patient-individual tumor models constitute a powerful platform for basic and translational analyses both in vitro and in vivo. However, due to the labor-intensive and highly time-consuming process, only few well-characterized patient-derived cell lines and/or corresponding xenografts exist. In this study, we describe successful generation and functional analysis of novel tumor models from patients with sporadic primary colorectal carcinomas (CRC) showing CpG island methylator phenotype (CIMP). Initial DNA fingerprint analysis confirmed identity with the patient in all four cases. These freshly established cells showed characteristic features associated with the CIMP-phenotype (HROC40: APCwt, TP53 mut, KRAS mut; 3/8 marker methylated; HROC43: APC mut, TP53 mut, KRAS mut; 4/8 marker methylated; HROC60: APCwt, TP53 mut, KRASwt; 4/8 marker methylated; HROC183: APC mut, TP53 mut, KRAS mut; 6/8 marker methylated). Cell lines were of epithelial origin (EpCAM+) with distinct morphology and growth kinetics. Response to chemotherapeutics was quite individual between cells, with stage I-derived cell line HROC60 being most susceptible towards standard clinically approved chemotherapeutics (e.g. 5-FU, Irinotecan). Of note, most cell lines were sensitive towards "non-classical" CRC standard drugs (sensitivity: Gemcitabin > Rapamycin > Nilotinib). This comprehensive analysis of tumor biology, genetic alterations and assessment of chemosensitivity towards a broad range of (chemo-) therapeutics helps bringing forward the concept of personalized tumor therapy.

Show MeSH
Related in: MedlinePlus