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PubMedDate not supplied

Thermal radiosensitization of human tumour cell lines with different sensitivities to 41.1 degrees C.

While much work on radiosensitization by hyperthermia in the 43 degrees C and higher temperature range has been done, relatively little work has been done at temperatures in the 41-42 degrees C range. In this moderate hyperthermia range there are dramatic differences in the resistance of mammalian cells to hyperthermia. Therefore, thermal radiosensitization was measured in two human colon adenocarcinoma cell lines, one that expresses chronic thermotolerance and proliferates at 41.1 degrees C, NSY 42129 (NSY) cells and one that is slowly killed at 41.1 degrees C, HCT15 cells. Heat-resistant NSY cells were found to be more radioresistant than heat-sensitive HCT15 cells. Hyperthermia at 41.1 degrees C enhanced the radiation sensitivity in NSY cells, but no significant induction of heat-induced radiosensitization was observed in HCT15 cells. The radiation sensitivity induced by 41.1 degrees C in NSY cells appeared to be related to both intrinsic heat-induced radiosensitization (HIR) and cell-cycle redistribution at 41.1 degrees C. Incidentally, cells incubated at 41.1 degrees C for between 8-16 h displayed an identical radiosensitivity to those heated for 24 h. This result implies that modest hyperthermia for 2 h or more can have a radiosensitizing effect in heat-resistant cells.

Adenocarcinoma↗

PubMed2001

Relationships between clonogenic cell survival, DNA damage and chromosomal radiosensitivity in nine human cervix carcinoma cell lines.

PURPOSE: To compare clonogenic cell survival, DNA damage and chromosomal radiosensitivity in nine cervix carcinoma cell lines. MATERIALS AND METHODS: Initial and residual (after 24h repair) radiation-induced DNA damage was evaluated using pulsed field gel electrophoresis. Chromosome damage was measured by micronucleus (MN) induction in cytochalasin-B-induced binucleate cells. RESULTS: Significant differences between the cell lines were obtained in the induced levels of initial damage, residual damage and MN. Values for SF2 for the nine cell lines ranged from 0.36 to 0.92. No correlation was found between clonogenic measurements of radiosensitivity and initial DNA damage dose response slopes. However, borderline significant correlations were seen between clonogenic radiosensitivity data and the levels of residual DNA damage. There was no correlation between clonogenic radiosensitivity and the levels of radiation-induced MN. Cell lines with high levels of initial damage had high yields of MN induced by radiation and the correlation seen was significant. CONCLUSIONS: The poor correlation between the different endpoints precludes their use in a clinical setting on primary tumour samples in vitro. It may be that tumour cell lines in vitro are a poor model for tumours in vivo. Studies aimed at assessing assays for measuring tumour radiosensitivity therefore should employ clinical samples. In vitro cell line work should concentrate on unravelling the complex mechanisms involved in determining a radiosensitive or radioresistant phenotype.

Cell Nucleus↗

PubMed2001

Chemical agents that promote chromatin compaction radiosensitize tumour cells.

PURPOSE: Previous studies indicated that cells whose chromatin is naturally compacted at the time of radiation are hypersensitive to radiation-induced killing, primarily by single-hit inactivation. Some chemicals that are known to promote chromatin compaction in interphase cells are here investigated for their radiosensitizing potential. MATERIALS AND METHODS: Okadaic acid (OA), a protein phosphatase inhibitor, fostriecin (FC), a topoisomerase II inhibitor and trichostatin A (TSA), a histone deacetylase inhibitor, were reported to promote chromatin compaction in mammalian cells. Asynchronous populations of HT-29 (human colon carcinoma) cells were exposed to various concentrations of OA, FC and TSA for various times before irradiation with various doses of Cs-137 gamma-rays and toxicity and radiosensitization were measured. Induced chromatin compaction was visualized by electron microscopy (EM). Histone 1 (H1) and histone 3 (H3) phosphorylation was measured by Western blotting, whole-cell fluorescence microscopy and confocal microscopy. RESULTS: OA and FC produced significant radiosensitization at 2 Gy after short (2 h) exposures. These chemical treatments also produced increased phosphorylation of H3 and increased chromatin compaction as measured by EM. A 2-h exposure of cells to TSA had no effect on cell radiosensitivity, histone phosphorylation or chromatin condensation. However, a 16-h exposure to TSA produced significant radiosensitization, histone phosphorylation and chromatin condensation, presumably by secondary mechanisms. CONCLUSIONS: These data are consistent with the hypothesis that compacted chromatin is a hypersensitive target for radiation killing. Furthermore, the modulation of chromatin conformation by drugs selectively in tumour cells might radiosensitize tumours whose cells are intrinsically radioresistant.

Alkenes↗

PubMed2002

Chromosome aberrations detected by FISH and correlation with cell survival after irradiation at various dose-rates and after bromodeoxyuridine radiosensitization.

PURPOSE: To determine whether measurement of chromosome aberrations by fluorescence in situ hybridization (FISH) predicts cell survival after irradiation at different dose-rates and after radiosensitization by bromodeoxyurdine (BrdU) in a lung carcinoma cell line. MATERIALS AND METHODS: The human lung carcinoma cell line SW1573 was irradiated at high dose-rate (HDR: 0.8 Gy min-1) or at pulsed low dose-rate (p-LDR: average dose-rate 1 Gy h-1) with or without radiosensitization by bromodeoxyuridine (BrdU). Cell survival was determined by clonogenic assay. Chromosome aberrations (colour junctions) were measured by whole-chromosome FISH of chromosome 2 and 18 and were scored according to the PAINT method. RESULTS: Clear radiosensitization by BrdU was observed both after HDR and p-LDR irradiation. Chromosome 18 was more radiosensitive than chromosome 2. There was a good correlation between induction of colour junctions and cell survival both after HDR and p-LDR irradiation and after radiosensitization by BrdU. CONCLUSIONS: Determination of chromosome aberrations by FISH can predict cell survival after different dose-rates and after radiosensitization by BrdU

Bromodeoxyuridine↗

PubMed2003

Influence of intrinsic radiosensitivity on the survival of breast cancer patients.

PURPOSE: The association between enhanced intrinsic radiosensitivity, measured by the level of chromosome damage induced in either the G2 or G0 phases of the cell cycle, and the risk of breast cancer has been well demonstrated. Although not originally designed for this purpose, early studies are now sufficiently mature (median follow-up 4.2 years) to enable an assessment of the impact of radiosensitivity on patient outcome. MATERIALS AND METHODS: Breast cancer patients assessed in previous studies of G2 and G0 lymphocyte chromosomal radiosensitivity between 1993 and 1997 were identified. Tumour characteristics, recurrence and survival data were obtained from clinical records and outcomes were compared between those identified as having normal and elevated radiosensitivity in the two assays. RESULTS: There was a significant association between G0 sensitivity and tumour grade (p=0.03), but otherwise there were no associations in these series between radiosensitivity in the two assays and tumour characteristics. Overall survival in this cohort of patients was high (94% at 5 years) and there was no association between the G2 or G0 assay results and outcome as measured by cancer-specific survival or local recurrence. CONCLUSIONS: Radiosensitivity, as measured by these two assays, was not a strong prognostic factor following standard radiotherapy treatment. However, the statistical power of the present study was limited and modest prognostic utility could not be discounted. Further studies designed specially to investigate outcome are needed, possibly in association with trials of altered radiotherapy fractionation.

Aged↗

PubMed1991

Trypsin-induced changes in cell shape and chromatin structure result in radiosensitization of monolayer Chinese hamster V79 cells.

Trypsin is the enzyme commonly used to prepare single cell suspensions from monolayer and spheroid cultures, both to determine survival and to assay DNA damage. Trypsin induces rounding, dissociation and radiosensitization of anchorage-dependent cells. Radiosensitivity and chromatin structure were compared between trypsin-treated (0.05%) round V79 cells from monolayers and spheroids vs. untreated spread monolayer cells in situ. The fluorescent halo technique was used to measure the changes in DNA supercoiling in nucleoids isolated from control and irradiated round and spread cells. Maximal halo diameters, the amount of initial and residual radiation-induced DNA damage (estimated from nucleoid halo diameter changes), and the radiosensitivity were higher in round cells than in spread monolayer V79 cells. The effects on cellular radiosensitivity and maximal halo diameter of other agents which also round and dissociate cells, e.g. 0.25% trypsin, pronase E and a non-enzymatic cell-dissociation solution, were similar to those of 0.05% trypsin. In LY-S cells, which are anchorage-independent, DNA loop size, the initial amount of DNA damage and radiosensitivity were not affected by trypsin. We suggest that the higher radiosensitivity of anchorage-dependent cells under immediate trypsinization and plating conditions, compared to cells with postirradiation in situ repair incubation, is due to correlated changes in cell shape and chromatin structure.

Animals↗

PubMed1992

Steepness of the clinical dose-control curve and variation in the in vitro radiosensitivity of head and neck squamous cell carcinoma.

Inter-tumour heterogeneity in radiobiological parameters has been proposed as an explanation for the quite shallow dose-response curves for local tumour control after radiotherapy observed in clinical data. Variability in the intrinsic radiosensitivity is potentially a very strong source of variation in local control. A method is presented for forcing such variability into a direct analysis (maximum-likelihood estimation) of tumour control data. The method is used to reanalyse a series of local tumour control data in 181 patients with squamous cell carcinoma of the oropharynx taking the distribution of in vitro radiosensitivities from an independent series of patients into account. It is concluded that direct application of the in vitro radiosensitivities leads to an unrealistically high estimate for the number of target cells per cm3. A more realistic fit is obtained after including a dose-modifying factor to correct for the apparent difference between in vitro and clinical radiosensitivities. The value of this factor is estimated at 2.4 with approximate 95% confidence interval (CI) (1.3, 5.9). It is suggested that hypoxia plays a role in reducing the radiosensitivity of tumours in clinical radiotherapy. Using this method provides more biologically reasonable estimates of other radiobiological parameters. The target-cell doubling time during treatment is estimated at 3.2 days with 95% CI (1.7, 8.7) days. Estimates of the target cell density in typical patients vary between 1.8 x 10(-6) and 6.6 x 10(-4) when the delay before accelerated tumour growth is assumed to vary between 0 and 28 days. Using the method presented here, the shallow clinical dose-control curve is interpreted as a superposition of quite steep dose-response relationships in individual patients. The steepness of the dose-control curve for a typical patient is characterized by a normalized dose-response gradient (the percentage change in tumour control for a 1% change in total dose) of 7.3 after stratification for intrinsic radiosensitivity as compared with 1.6 if such stratification is not performed.

Carcinoma, Squamous Cell↗

PubMed1994

Nicotinamide and carbogen: relationship between pO2 and radiosensitivity in three tumour lines.

The effects of carbogen breathing, nicotinamide injection and their combination on tumour radiosensitivity were correlated with changes in tumour O2 tension to determine the relationship between radiosensitivity and measured pO2. The radiosensitivity (in vivo-in vitro colony assay) and O2 tension (computerized pO2 histograph KIMOC 6650) of two human xenografted tumours (HRT18 and Na11+) and one murine tumour (EMT6) were measured under similar experimental conditions. A single dose of radiation was delivered (8 Gy for HRT18 and Na11+, 12 Gy for EMT6). Carbogen breathing, nicotinamide injection, and their combination all significantly increased radiosensitivity in the three cell lines (p < 0.05); the most efficient treatment was carbogen plus nicotinamide. The radiosensitization was optimum for EMT6 and Na11+. Mean and median pO2 increased with all three treatments, except for carbogen breathing in EMT6. Carbogen breathing had little effect on the proportion of low pO2 values, but induced pO2 values > 30 mmHg in all three tumour lines. Nicotinamide decreased the proportion of low pO2 values. This effect is larger with the combination carbogen plus nicotinamide. Almost all pO2 values < 2.5 mmHg were eliminated for HRT18 and EMT6. The relationship between radiosensitization and pO2 was significant when pO2 was expressed as the percentage of values below 7, 8, 9 and 10 mmHg for the three lines.

Animals↗

PubMed1994

Use of fluorescence in situ hybridization to measure chromosome aberrations as a predictor of radiosensitivity in human tumour cells.

Fluorescence in situ hybridization (FISH) is a potential assay for determining cellular radiosensitivity based on the detection of chromosome damage. This approach was chosen because of its relative simplicity and short assay time. Two radiosensitive and two radioresistant human tumour cell lines were used. The radiosensitive lines were an ovarian carcinoma line (A1847) and a squamous carcinoma line (SCC61). The radioresistant cells were a lung adenocarcinoma line (A549) and a second squamous line (SQ20B). Whole chromosome-specific probes were used to detect radiation-induced chromosome aberrations in mitotic cells. Available probes were first screened to characterize the intrinsic chromosome aberrations before irradiation and the appropriate probes (minimum fluorescent spots) were selected for each cell line. Maximum radiation-induced aberrations were found 24 h after irradiation. Dose-response curves corrected for target size (proportion of genome probed) differed for all cell lines. The radiosensitive A1847 cell line showed more induced aberrations compared with the radioresistant A549 cell line, in agreement with the survival data. In contrast, the SQ20B cell line showed more induced chromosome aberrations than the more radiosensitive SCC61 cell line, leading to the hypothesis that the SQ20B cells could tolerate more aberrations. Dose-response curves obtained in surviving cells 14 days postirradiation indeed showed elevated levels of chromosome aberrations for SQ20B cells. The difference in chromosome aberrations between 1 and 14 days showed a good correlation with the survival data for all four cell lines. In conclusion, FISH of mitotic cells with whole chromosome probes appears to be a suitable assay to predict radiosensitivity. It seems necessary, however, to determine both induced and remaining chromosome aberrations, since different processing or tolerance of radiation-induced aberrations, including stable types, could lead to different correlations with cell survival.

Cell Survival↗

PubMed1999

Chromosome radiosensitivity in human G2 lymphocytes and cell-cycle progression.

PURPOSE: To investigate the possibility that the differential G2-phase radiosensitivity of human peripheral blood lymphocytes, found in normal individuals using the 'G2-phase chromosome radiosensitivity assay', could be attributed to heterogeneity in cellular progression to mitosis rather than differences in radiosensitivity. MATERIALS AND METHODS: Human peripheral blood lymphocytes, from four different donors, were exposed to 50 cGy X-rays and sampled at different times. The progression of cells into mitosis was monitored by 5-bromo 2'-deoxyuridine (BrdUrd) incorporation. RESULTS: The heterogeneous G2-phase chromosome radiosensitivity among different donors was abolished when homogeneous G2-phase cell populations were scored; they contained similar frequencies of cells in early or late G2-phase. CONCLUSIONS: The heterogeneous G2-phase chromosome radiosensitivity, usually found in different normal donors, is caused by the analysis of different cell populations rather than reflecting intrinsic differences in radiosensitivity.

Adult↗

PubMed1999

Heritability of cellular radiosensitivity: a marker of low-penetrance predisposition genes in breast cancer?

Many inherited cancer-prone conditions show an elevated sensitivity to the induction of chromosome damage in cells exposed to ionizing radiation, indicative of defects in the processing of DNA damage. We earlier found that 40% of patients with breast cancer and 5%-10% of controls showed evidence of enhanced chromosomal radiosensitivity and that this sensitivity was not age related. We suggested that this could be a marker of cancer-predisposing genes of low penetrance. To further test this hypothesis, we have studied the heritability of radiosensitivity in families of patients with breast cancer. Of 37 first-degree relatives of 16 sensitive patients, 23 (62%) were themselves sensitive, compared with 1 (7%) of 15 first-degree relatives of four patients with normal responses. The distribution of radiosensitivities among the family members showed a trimodal distribution, suggesting the presence of a limited number of major genes determining radiosensitivity. Segregation analysis of 95 family members showed clear evidence of heritability of radiosensitivity, with a single major gene accounting for 82% of the variance between family members. The two alleles combine in an additive (codominant) manner, giving complete heterozygote expression. A better fit was obtained to a model that includes a second, rarer gene with a similar, additive effect on radiosensitivity, but the data are clearly consistent with a range of models. Novel genes involved in predisposition to breast cancer can now be sought through linkage studies using this quantitative trait.

Alleles↗

PubMed1992

Differential expression of proteins in radioresistant and radiosensitive human squamous carcinoma cells.

BACKGROUND: Previous studies support a genetic basis for cellular radioresistance. The associated biochemical and molecular events, however, are not fully understood. PURPOSE: We investigated the differential protein pattern as a molecular determinant of resistance or sensitivity of head and neck squamous carcinoma cells to ionizing radiation. METHODS: Using two-dimensional polyacrylamide gel electrophoresis followed by computer-assisted quantitative analysis, we compared the protein profiles of three relatively radioresistant and three relatively radiosensitive head and neck squamous carcinoma cell lines (previously characterized by in vitro and clinical parameters as radioresistant or radiosensitive) to determine which proteins were consistently expressed or enhanced in the radioresistant compared with the radiosensitive phenotype. RESULTS: Our analysis indicated that 14 proteins were preferentially expressed in the radio-resistant cell lines SQ-20B, JSQ-3, and SCC-35, with one protein (molecular mass of 92 kd and pI of 5.5) distinctly expressed in the radioresistant cell lines. Four proteins were enhanced by greater than 10-fold, three were enhanced fivefold to 10-fold, and six were enhanced twofold to fivefold in the radioresistant cell lines. In addition, we observed a second set of 15 proteins preferentially expressed in the radio-sensitive cell lines SQ-9G, SQ-38, and SCC-9. A 40-kd protein (pI 7.1) was distinctly expressed in the radiosensitive cell lines. The remaining radiosensitive cell-specific proteins were enhanced by greater than 10-fold (two proteins), fivefold to 10-fold (two proteins), or twofold to fivefold (10 proteins) compared with their counterparts in the radioresistant cell lysates. CONCLUSION: These results provide evidence for differential protein expression associated with phenotypic expression of cellular radioresistance or radiosensitivity. IMPLICATIONS: This study will facilitate the characterization of these proteins correlated with the radiation response-specific phenotype.

Carcinoma, Squamous Cell↗

PubMed1996

Radiosensitizing nucleosides.

Chemotherapeutic drugs that perturb nucleotide metabolism have the potential to produce substantial sensitization of tumor cells to radiation treatment. The process is called radiosensitization, and the agents that induce it are called radiosensitizers. The clinical effectiveness of fluoropyrimidines as radiosensitizers has been proven in multiple randomized trials. Thymidine analogues and hydroxyurea also appear to produce clinically relevant increases in radiation sensitivity. Recent laboratory investigations have identified difluorodeoxycytidine (gemcitabine) and fludarabine as promising agents to use in combination with radiation. Until recently, little was known about how the biochemical changes caused by these drugs produced radiosensitization. However, advances in related fields, such as cell cycle checkpoint control, have permitted the development of a hypothesis that may explain the relative tumor selectivity of fluoropyrimidine-mediated radiosensitization. In addition, recent findings suggest that the rational manipulation of drug administration schedules and the use of combinations of radiosensitizers have the potential to improve the efficacy of the currently used agents and to establish the benefit of new ones.

Animals↗

PubMed1981

Huntington's disease: implications of associated cellular radiosensitivity.

Ionizing radiation sensitivity was studied in a series of Huntington's Disease (HD) patients and controls by measurement of radiation-induced chromosome aberrations in lymphocytes and by clonogenic survival of lymphoblastoid cell lines. As a group, HD patients were found to be significantly more radiosensitive than controls (p less than 0.001), but there was an overlap between values for the two groups such that an absolute distinction is not possible. These data are consistent with an association between HD and radiosensitivity but not with identity between HD and a radiosensitive phenotype, so that cellular radiosensitivity cannot be used for individual diagnosis. Analysis of three families including 5 HD patients and 11 first-degree relative confirmed this conclusion and demonstrated that even within a given family presymptomatic diagnosis cannot be based on measurement of radiosensitivity. However, the common association of cellular radiosensitivity with HD probands and their families provides a potential lead to the identification of HD gene(s) and so to an eventual understanding of the aetiopathogenesis of this disease at the molecular level.

Cell Line↗

PubMed2002

Radiation could induce p53-independent and cell cycle--unrelated apoptosis in 5-fluorouracil radiosensitized head and neck carcinoma cells.

The effect of chemoresistance induction in radiosensitivity and cellular behavior after irradiation remains misunderstood. This study was designed to understand the relationship between radiation-induced cell cycle arrest, apoptosis, and radiosensitivity in KB cell line and KB3 subline selected after 5-fluorouracil (5FU) exposure. Exposure of KB cells to 5FU led to an increase in radiosensitivity. G2/M cell cycle arrest was observed in the two cell lines after irradiation. The radioresistant KB cell line reached the maximum arrest two hours before KB3. The cellular exit from this arrest was found to be related to the wild type p53 protein expression induction. After irradiation, only KB3 cell line underwent apoptosis. This apoptosis induction seemed to be independent of G2/M arrest exit, which was carried out later. The difference in radiosensitivity between KB and KB3 subline may result therefore from both a difference in apoptosis induction and a difference in G2/M arrest maximum duration. Moreover, 5FU exposure has led to an increase in constitutive p53 protein expression, which may be associated with an increase in basal apoptosis cell fraction. Given the existing correlation between radiosensitivity and the percentage of basal apoptosis, the constitutive p53 protein expression may be related to intrinsic radiosensitivity in our cellular model.

Annexin A5↗

PubMed2006

Tumor cell radiosensitivity is a major determinant of tumor response to radiation.

Substantial evidence suggests that the radiosensitivity of the tumor cells is the primary determinant of tumor response to radiation. More recent studies suggest that tumor stroma radiosensitivity is the principle determinant of response. To assess the relationship between intrinsic tumor cell radiosensitivity and tumor response, we altered the intrinsic radiosensitivity of a cloned tumor cell line and analyzed the effect of this alteration on tumor response. A cloned tumor cell line derived from DNA double-strand break repair--deficient severe combined immunodeficient mice was transfected with the double-strand break repair gene DNA-PKcs. The intrinsic radiosensitivity of the transfected tumor line was decreased by a factor of approximately 1.5. The isogenic lines were used to initiate tumors in NCr-nu/nu mice. When transplanted in the same strain of mice and exposed to the same dose of radiation, the isogenic tumors may be expected to exhibit a similar response to radiation if radiation damage to host stroma is the principle determinant of response. This was not observed. Over the dose range of 20 Gy in four 5-Gy fractions to a single dose of 30 Gy, the 1.5-fold increase in intrinsic tumor cell radioresistance conferred by the introduction of DNA-PKcs caused a 1.5-fold decrease in tumor growth delay. The results show that the intrinsic radiosensitivity of tumor cells is a major determinant of tumor response to radiation.

Animals↗

PubMed2004

Enhanced tumor cell radiosensitivity and abrogation of G2 and S phase arrest by the Hsp90 inhibitor 17-(dimethylaminoethylamino)-17-demethoxygeldanamycin.

PURPOSE: Because of the potential for affecting multiple signaling pathways, inhibition of Hsp90 may provide a strategy for enhancing tumor cell radiosensitivity. Therefore, we have investigated the effects of the orally bioavailable Hsp90 inhibitor 17-(dimethylaminoethylamino)-17-demethoxygeldanamycin (17-DMAG) on the radiosensitivity of human tumor cells in vitro and grown as tumor xenografts. EXPERIMENTAL DESIGN: The effect of 17-DMAG on the levels of three proteins (Raf-1, ErbB2, and Akt) previously implicated in the regulation of radiosensitivity was determined in three human solid tumor cell lines. A clonogenic assay was then used to evaluate cell survival after exposure to 17-DMAG followed by irradiation. For mechanistic insight, the G(2)- and S-phase checkpoints were evaluated in 17-DMAG-treated cells. Finally, the effect of in vivo administration of 17-DMAG in combination with radiation on the growth rate of xenograft tumors was determined. RESULTS: 17-DMAG exposure reduced the levels of the three radiosensitivity-associated proteins in a cell line-specific manner with ErbB2 being the most susceptible. Corresponding concentrations of 17-DMAG enhanced the radiosensitivity of each of the tumor cell lines. This sensitization seemed to be the result of a 17-DMAG-mediated abrogation of the G(2)- and S-phase cell cycle checkpoints. The oral administration of 17-DMAG to mice bearing tumor xenografts followed by irradiation resulted in a greater than additive increase in tumor growth delay. CONCLUSIONS: These data indicate that 17-DMAG enhances the in vitro and in vivo radiosensitivity of human tumor cells. The mechanism responsible seems to involve the abrogation of radiation-induced G(2)- and S-phase arrest.

Administration, Oral↗

PubMed2006

2-methoxyestradiol, an endogenous mammalian metabolite, radiosensitizes colon carcinoma cells through c-Jun NH2-terminal kinase activation.

PURPOSE: 2-Methoxyestradiol (2ME), an estrogen metabolite, induces apoptosis in various cell types. We investigated whether 2ME pretreatment can radiosensitize colon adenocarcinoma cells. EXPERIMENTAL DESIGN: Radiosensitizing effects of 2ME were evaluated by cell death, clonogenic assay, nuclear fragmentation, and tumor progression of xenografts. Ionizing radiation-induced DNA damage was evaluated by histone H2AX phosphorylation and its foci. The c-Jun NH2-terminal kinase (JNK) activation was evaluated by anti-phosphorylated JNK antibody and inhibited by the JNK-specific inhibitor SP600125 or dominant-negative SEK1 expression. RESULTS: Clonogenic assays revealed that 2ME, but not estradiol, radiosensitized three colon carcinoma cells, DLD-1, HCT-8, and HCT-15, and strongly suppressed tumor progression of DLD-1 xenografts. Gene transfer-mediated Bcl-xL overexpression largely abolished both augmented apoptosis and reduced survival fractions. Pretreatment with 2ME enhanced H2AX phosphorylation, its foci, and phosphorylation of ATM kinase and delayed re-entry of cell cycle progression after ionizing radiation. Augmentation of both radiosensitivity and H2AX phosphorylation was substantially reduced by SP600125 or overexpression of a dominant-negative mutant SEK1. CONCLUSION: 2ME radiosensitized colon carcinoma cells through enhanced DNA damage via JNK activation, thereby representing a novel radiosensitizing therapy against colon cancer.

2-Methoxyestradiol↗