Soft agarose culture human tumour colony forming assay for drug sensitivity testing: [3H]-thymidine incorporation vs colony counting

Jones, CM; Tsukamoto, Taiji; O’Brien, Patrícia C. M.; Uhl, Cindy B.; Alley, Michael C.; Lieber, Michael M.

doi:10.1038/bjc.1985.194

Cited by 24 publications

(6 citation statements)

References 16 publications

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“…Colony formation of human lung cancer cells in soft agar was performed, as previously described (34), with 2,500 cells seeded in 6-well plate in DMEM with 10% FBS. Colonies were counted after 2 weeks.…”

Section: Methodsmentioning

confidence: 99%

Multivalent Forms of the Notch Ligand DLL-1 Enhance Antitumor T-cell Immunity in Lung Cancer and Improve Efficacy of EGFR-Targeted Therapy

et al. 2015

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Activation of Notch signaling in hematopoietic cells by tumors contributes to immune escape. T cell defects in tumors can be reversed by treating tumor-bearing mice with multivalent forms of the Notch receptor ligand DLL-1, but the immunologic correlates of this effect have not been elucidated. Here we report mechanistic insights along with the efficacy of combinational treatments of multivalent DLL-1 with oncoprotein targeting drugs in preclinical mouse models of lung cancer. Systemic DLL-1 administration increased T cell infiltration into tumors and elevated numbers of CD44+CD62L+CD8+ memory T cells, while decreasing the number of regulatory T cells and limiting tumor vascularization. This treatment was associated with upregulation of Notch and its ligands in tumor-infiltrating T cells, enhanced expression of T-bet and phosphorylation of Stat1/2. Adoptive transfer of T cells from DLL1-treated tumor-bearing immunocompetent hosts into tumor-bearing SCID-NOD immunocompromised mice attenuated tumor growth and extended tumor-free survival in the recipients. When combined with the EGFR-targeted drug erlotinib, DLL-1 significantly improved progression-free survival by inducing robust tumor-specific T cell immunity. In tissue culture, DLL1 induced proliferation of human peripheral T cells but lacked proliferative or clonogenic effects on lung cancer cells. Our findings offer preclinical mechanistic support for the development of multivalent DLL1 to stimulate anti-tumor immunity.

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Section: Methodsmentioning

confidence: 99%

Multivalent Forms of the Notch Ligand DLL-1 Enhance Antitumor T-cell Immunity in Lung Cancer and Improve Efficacy of EGFR-Targeted Therapy

et al. 2015

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“…The influence of GCS on cell growth in soft agar was analyzed by [ 3 H]thymidine incorporation (15,16). Cells (2ϫ10 4 ) were suspended in 0.5 ml RPMI 1640 medium (supplemented with 2 U/ml insulin and 10% FBS), containing 0.25% agarose III and the indicated concentrations of adriamycin.…”

Section: Colony Formation In Soft Agarmentioning

confidence: 99%

Ceramide glycosylation potentiates cellular multidrug resistance

et al. 2001

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Ceramide glycosylation, through glucosylceramide synthase (GCS), allows cellular escape from ceramide-induced programmed cell death. This glycosylation event confers cancer cell resistance to cytotoxic anticancer agents [Liu, Y. Y., Han, T. Y., Giuliano, A. E., and M. C. Cabot. (1999) J. Biol. Chem. 274, 1140-1146]. We previously found that glucosylceramide, the glycosylated form of ceramide, accumulates in adriamycin-resistant breast carcinoma cells, in vinblastine-resistant epithelioid carcinoma cells, and in tumor specimens from patients showing poor response to chemotherapy. Here we show that multidrug resistance can be increased over baseline and then totally reversed in human breast cancer cells by GCS gene targeting. In adriamycin-resistant MCF-7-AdrR cells, transfection of GCS upgraded multidrug resistance, whereas transfection of GCS antisense markedly restored cellular sensitivity to anthracyclines, Vinca alkaloids, taxanes, and other anticancer drugs. Sensitivity to the various drugs by GCS antisense transfection increased 7- to 240-fold and was consistent with the resumption of ceramide-caspase-apoptotic signaling. GCS targeting had little influence on cellular sensitivity to either 5-FU or cisplatin, nor did it modify P-glycoprotein expression or rhodamine-123 efflux. GCS antisense transfection did enhance rhodamine-123 uptake compared with parent MCF-7-AdrR cells. This study reveals that GCS is a novel mechanism of multidrug resistance and positions GCS antisense as an innovative force to overcome multidrug resistance in cancer chemotherapy.

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“…In vitro test systems aid in predicting antitumor drug effects [2,3]. SD is an ATP-producing key enzyme of the citric cycle and as the activity of this enzyme correlates well with the cell viability, it can be used to predict the clinical response [5,6], The SD1 test has not been popular compared with the clonogenic assay or isotope incorporation method for in vitro chemosensitivity testing [2], However the SD1 test is a simple, inexpensive and rapid technique for screen ing antitumor drugs.…”

Section: Discussionmentioning

confidence: 99%

“…In vitro and in vivo trials for predicting the che mosensitivity of individual tumors have been reported [1][2][3]. The in vitro succinate dehydrogenase inhibition (SDI) test was introduced for chemosensitivity testing, based on the correlation of succinate dehydrogenase (SD; EC 1.3.99.1) activity using tetrazolium salt as a hydrogen acceptor, with cell viability [4].…”

Section: Introductionmentioning

confidence: 99%

Comparison between Succinate Dehydrogenase Inhibition Test and Subrenal Capsule Assay for Chemosensitivity Testing

Anai

Maehara²,

Kusumoto

et al. 1987

Oncology

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The chemosensitivity result of the succinate dehydrogenase inhibition (SDI) test was compared with that of the subrenal capsule (SRC) assay in 23 human tumor tissues exposed to adriamycin (ADM), mitomycin C (MMC), cisplatin (DDP) and 5-fluorouracil (5-FU). The chemosensitivity was considered as positive when the succinate dehydrogenase (SD) activity of the drug-exposed cells was decreased to below 50% of that of control cells on day 3 in the SDI test, and the tumor size on day 6 was decreased to below 10% of that on day 0 in the SRC assay. Correlation rates between the decrease of SD activity in the SDI test and the decrease of tumor size in the SRC assay, using 23 evaluable cases in both assays, were r = 0.717 for ADM, r = 0.699 for MMC, r = 0.796 for DDP and r = 0.735 for 5-FU. The correlations of the chemosensitivity results were 73.9% for ADM, 73.9% for MMC, 82.6% for DDP and 60.9% for 5-FU. A positive correlation was noted between the in vitro and in vivo chemosensitivity results. This SDI test can serve as an effective tool for chemosensitivity testing.

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Soft agarose culture human tumour colony forming assay for drug sensitivity testing: [3H]-thymidine incorporation vs colony counting

Cited by 24 publications

References 16 publications

Multivalent Forms of the Notch Ligand DLL-1 Enhance Antitumor T-cell Immunity in Lung Cancer and Improve Efficacy of EGFR-Targeted Therapy

Multivalent Forms of the Notch Ligand DLL-1 Enhance Antitumor T-cell Immunity in Lung Cancer and Improve Efficacy of EGFR-Targeted Therapy

Ceramide glycosylation potentiates cellular multidrug resistance

Comparison between Succinate Dehydrogenase Inhibition Test and Subrenal Capsule Assay for Chemosensitivity Testing

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