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In vitro sensitivity testing of minimally passaged and uncultured gliomas with TRAIL and/or chemotherapy drugs.

Ashley DM, Riffkin CD, Lovric MM, Mikeska T, Dobrovic A, Maxwell JA, Friedman HS, Drummond KJ, Kaye AH, Gan HK, Johns TG, Hawkins CJ - Br. J. Cancer (2008)

Bottom Line: Cross-linked TRAIL, but not soluble TRAIL, killed both normal cell types and cells from three tumours.This study highlights the widespread resistance of glioma cells to TRAIL-based agents, but suggests that a minority of high-grade glioma patients may benefit from particular combinations of TRAIL and chemotherapy drugs.In vitro sensitivity assays may help identify effective drug combinations for individual glioma patients.

View Article: PubMed Central - PubMed

Affiliation: Children's Cancer Centre, Murdoch Children's Research Institute, Royal Children's Hospital, Parkville, Victoria 3052, Australia.

ABSTRACT
TRAIL/Apo-2L has shown promise as an anti-glioma drug, based on investigations of TRAIL sensitivity in established glioma cell lines, but it is not known how accurately TRAIL signalling pathways of glioma cells in vivo are reproduced in these cell lines in vitro. To replicate as closely as possible the in vivo behaviour of malignant glioma cells, 17 early passage glioma cell lines and 5 freshly resected gliomas were exposed to TRAIL-based agents and/or chemotherapeutic drugs. Normal human hepatocytes and astrocytes and established glioma cell lines were also tested. Cross-linked TRAIL, but not soluble TRAIL, killed both normal cell types and cells from three tumours. Cells from only one glioma were killed by soluble TRAIL, although only inefficiently. High concentrations of cisplatin were lethal to glioma cells, hepatocytes and astrocytes. Isolated combinations of TRAIL and chemotherapy drugs were more toxic to particular gliomas than normal cells, but no combination was generally selective for glioma cells. This study highlights the widespread resistance of glioma cells to TRAIL-based agents, but suggests that a minority of high-grade glioma patients may benefit from particular combinations of TRAIL and chemotherapy drugs. In vitro sensitivity assays may help identify effective drug combinations for individual glioma patients.

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Related in: MedlinePlus

Propidium iodide uptake assay of glioma cell sensitivity to combination treatments. Cells from the indicated early passage glioma cell lines or ex vivo gliomas were incubated in vitro with soluble TRAIL at 100 ng ml−1 (+) or 1000 ng ml−1 (++) alone or together with temozolomide (13.7 μg ml−1) or cisplatin (54 μg ml−1) for 48 h. Flow cytometry measurement of propidium iodide exclusion was used to quantitate the proportion of surviving cells. The areas of the circles denote survival following each treatment. Small circles indicate efficient killing, large circles reflect survival.
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fig4: Propidium iodide uptake assay of glioma cell sensitivity to combination treatments. Cells from the indicated early passage glioma cell lines or ex vivo gliomas were incubated in vitro with soluble TRAIL at 100 ng ml−1 (+) or 1000 ng ml−1 (++) alone or together with temozolomide (13.7 μg ml−1) or cisplatin (54 μg ml−1) for 48 h. Flow cytometry measurement of propidium iodide exclusion was used to quantitate the proportion of surviving cells. The areas of the circles denote survival following each treatment. Small circles indicate efficient killing, large circles reflect survival.

Mentions: Previous studies have shown that co-treatment with traditional anti-cancer agents can sensitise some cells to TRAIL, in vitro and in vivo. We tested the ability of the various TRAIL formulations to kill glioma cells in conjunction with a number of chemotherapy drugs used in glioma therapy (Table 2). Figure 2 illustrates the effect of these combination treatments on D2247 (the most TRAIL-sensitive line), D2235 and D2248 (two of the TRAIL resistant lines) and RMH020 (one of the uncultured tumours). Average responses across all early passage lines and ex vivo tumours are shown in Figure 3. Many samples were killed by high-dose cisplatin, however a 10-fold lower concentration was much less effective. The higher dose of vincristine was weakly toxic to many of the glioma samples. Only the D2235 cells were markedly sensitive to temozolomide. The other drugs were ineffective as sole agents. All chemotherapy drugs tested further sensitised D2247 to F-LZ-TRAIL and anti-DR5 (Figure 2), but only additive toxicity was observed when TRAIL-resistant cells were exposed to the combination treatments (Figures 2 and 3 and data not shown). The possibility that prior treatment with chemotherapy drugs may enhance sensitivity to TRAIL was also explored. Cells from a TRAIL-resistant early passage line, D2302, were incubated with chemotherapy drugs only for 24 h, then TRAIL or anti-DR5 antibody was added for an additional 48 h period. These treatments had similar effects on cellular ATP levels to co-incubations with TRAIL plus chemotherapy drugs for either 48 or 72 h (Supplementary Figure 1), indicating that prior exposure to chemotherapy drugs did not sensitise D2302 cells to TRAIL. Propidium iodide uptake assays were performed on many of the samples, using selected drug combinations. This method, which gives a direct measure of the proportion of cells killed, yielded similar data to the CellTiter-Glo assay, which quantifies cellular ATP (Figure 4).


In vitro sensitivity testing of minimally passaged and uncultured gliomas with TRAIL and/or chemotherapy drugs.

Ashley DM, Riffkin CD, Lovric MM, Mikeska T, Dobrovic A, Maxwell JA, Friedman HS, Drummond KJ, Kaye AH, Gan HK, Johns TG, Hawkins CJ - Br. J. Cancer (2008)

Propidium iodide uptake assay of glioma cell sensitivity to combination treatments. Cells from the indicated early passage glioma cell lines or ex vivo gliomas were incubated in vitro with soluble TRAIL at 100 ng ml−1 (+) or 1000 ng ml−1 (++) alone or together with temozolomide (13.7 μg ml−1) or cisplatin (54 μg ml−1) for 48 h. Flow cytometry measurement of propidium iodide exclusion was used to quantitate the proportion of surviving cells. The areas of the circles denote survival following each treatment. Small circles indicate efficient killing, large circles reflect survival.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2480982&req=5

fig4: Propidium iodide uptake assay of glioma cell sensitivity to combination treatments. Cells from the indicated early passage glioma cell lines or ex vivo gliomas were incubated in vitro with soluble TRAIL at 100 ng ml−1 (+) or 1000 ng ml−1 (++) alone or together with temozolomide (13.7 μg ml−1) or cisplatin (54 μg ml−1) for 48 h. Flow cytometry measurement of propidium iodide exclusion was used to quantitate the proportion of surviving cells. The areas of the circles denote survival following each treatment. Small circles indicate efficient killing, large circles reflect survival.
Mentions: Previous studies have shown that co-treatment with traditional anti-cancer agents can sensitise some cells to TRAIL, in vitro and in vivo. We tested the ability of the various TRAIL formulations to kill glioma cells in conjunction with a number of chemotherapy drugs used in glioma therapy (Table 2). Figure 2 illustrates the effect of these combination treatments on D2247 (the most TRAIL-sensitive line), D2235 and D2248 (two of the TRAIL resistant lines) and RMH020 (one of the uncultured tumours). Average responses across all early passage lines and ex vivo tumours are shown in Figure 3. Many samples were killed by high-dose cisplatin, however a 10-fold lower concentration was much less effective. The higher dose of vincristine was weakly toxic to many of the glioma samples. Only the D2235 cells were markedly sensitive to temozolomide. The other drugs were ineffective as sole agents. All chemotherapy drugs tested further sensitised D2247 to F-LZ-TRAIL and anti-DR5 (Figure 2), but only additive toxicity was observed when TRAIL-resistant cells were exposed to the combination treatments (Figures 2 and 3 and data not shown). The possibility that prior treatment with chemotherapy drugs may enhance sensitivity to TRAIL was also explored. Cells from a TRAIL-resistant early passage line, D2302, were incubated with chemotherapy drugs only for 24 h, then TRAIL or anti-DR5 antibody was added for an additional 48 h period. These treatments had similar effects on cellular ATP levels to co-incubations with TRAIL plus chemotherapy drugs for either 48 or 72 h (Supplementary Figure 1), indicating that prior exposure to chemotherapy drugs did not sensitise D2302 cells to TRAIL. Propidium iodide uptake assays were performed on many of the samples, using selected drug combinations. This method, which gives a direct measure of the proportion of cells killed, yielded similar data to the CellTiter-Glo assay, which quantifies cellular ATP (Figure 4).

Bottom Line: Cross-linked TRAIL, but not soluble TRAIL, killed both normal cell types and cells from three tumours.This study highlights the widespread resistance of glioma cells to TRAIL-based agents, but suggests that a minority of high-grade glioma patients may benefit from particular combinations of TRAIL and chemotherapy drugs.In vitro sensitivity assays may help identify effective drug combinations for individual glioma patients.

View Article: PubMed Central - PubMed

Affiliation: Children's Cancer Centre, Murdoch Children's Research Institute, Royal Children's Hospital, Parkville, Victoria 3052, Australia.

ABSTRACT
TRAIL/Apo-2L has shown promise as an anti-glioma drug, based on investigations of TRAIL sensitivity in established glioma cell lines, but it is not known how accurately TRAIL signalling pathways of glioma cells in vivo are reproduced in these cell lines in vitro. To replicate as closely as possible the in vivo behaviour of malignant glioma cells, 17 early passage glioma cell lines and 5 freshly resected gliomas were exposed to TRAIL-based agents and/or chemotherapeutic drugs. Normal human hepatocytes and astrocytes and established glioma cell lines were also tested. Cross-linked TRAIL, but not soluble TRAIL, killed both normal cell types and cells from three tumours. Cells from only one glioma were killed by soluble TRAIL, although only inefficiently. High concentrations of cisplatin were lethal to glioma cells, hepatocytes and astrocytes. Isolated combinations of TRAIL and chemotherapy drugs were more toxic to particular gliomas than normal cells, but no combination was generally selective for glioma cells. This study highlights the widespread resistance of glioma cells to TRAIL-based agents, but suggests that a minority of high-grade glioma patients may benefit from particular combinations of TRAIL and chemotherapy drugs. In vitro sensitivity assays may help identify effective drug combinations for individual glioma patients.

Show MeSH
Related in: MedlinePlus