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G D Wilson

Publications and source records attributed to G D Wilson.

At least 73 records · Page 4Linked to original sources

Flow cytometric analysis of p53 oncoprotein expression in cutaneous melanoma.

BACKGROUND: Mutation of the p53 tumour suppressor gene is thought to represent a significant step in the development of over half of all human cancers. Qualitative investigations of p53 protein in cutaneous melanoma suggest that overexpression is more common than in other tumours. This study analysed p53 protein expression using flow cytometry as a quantitative assay. METHODS: Expression of p53 protein was assayed in ten benign melanocytic naevi (BMN) and 50 surgically excised ethanol-fixed melanomas. Nuclei were stained for the p53 protein with Pab1801 and analysed using flow cytometry. Protein expression was further correlated with clinicopathological parameters. RESULTS: None of the BMN was immunopositive for p53. Forty-one of the 50 melanomas stained positively. The overall median positivity for p53 in primary melanoma was 12.8 per cent compared with 22.5 per cent in metastatic disease. No association was found between p53 expression and Breslow thickness or presence of nodal and skin metastases. Overexpression of p53 was associated with ulceration, mitotic figures and lymphocytic infiltration, and there was a striking increase of p53 expression with age. Expression of p53 was inversely correlated with disease-free interval. CONCLUSION: Although overexpression of p53 is common in melanoma, there was no association with major clinical parameters such as Breslow thickness and overall survival. The association found between p53 and other clinicopathological features suggests that the tumour suppressor gene does play a role in the evolution of melanoma.

Adult↗

Cell proliferation in renal cell carcinoma--a comparative study of cell kinetic methods.

Four different methods of assessing cell proliferation in renal cell carcinoma were compared in a total of 136 samples to analyze their degree of agreement and usefulness. The methods compared were flow cytometric S-phase (S-FCM) analysis, proliferating cell nuclear antigen expression detected by immunohistochemistry (PCNA-IHC), in vivo iododeoxyuridine incorporation analyzed with immunohistochemistry (IdUrd-IHC), and flow cytometry (IdUrd-FCM). The mean S-FCM fraction was 5.9%, compared with a mean PCNA-IHC labeling index of 4.7%. The mean labeling indices obtained by IdUrd-IHC and IdUrd-FCM were 1.2% and 1.7%, respectively. The four methods correlated well with each other. When the methods were compared according to Bland and Altman, good agreement was shown. A statistically significant difference in proliferation between diploid and aneuploid tumor samples was found with all methods (P < 0.001). The results showed that the four different methods provided comparable information on proliferative activity, although different cell cycle compartments were monitored.

Aneuploidy↗

C-myc oncogene expression in human melanoma and its relationship with tumour antigenicity.

Melanoma produces specific tumour antigens which are capable of eliciting an immune response. However, this tumour evades the immune system, in part, by downregulation of class I HLA antigens on the cell surface, which are required for T cell recognition. It has been suggested that the oncogene c-myc may have a role in effecting this change in vitro, however, the relationship between oncoprotein level and tumour antigenicity has not been established in human tumours. This study measured c-myc oncoprotein in 94 melanoma specimens (46 primary tumours and 48 regional metastases) using flow cytometry and evaluated class I HLA expression with immunohistochemistry. C-myc expression was found in 91 tumours (96%) with higher expression in metastases than primary melanomas (P<0.005). Class I HLA expression was found to show great variation although metastases showed less antigenicity than primary tumours (P<0.01). Analysis of the relationship between these two parameters revealed a highly significant correlation in both primary (P<0.01) and metastatic disease (P<0.01), with high oncoprotein being associated with down regulation of cell surface antigens. Knowledge of the control of tumour antigenicity is likely to provide an objective platform for the development of new strategies for immunotherapy.

Antigens, Neoplasm↗

Bcl-2 expression in malignant melanoma and its prognostic significance.

Programmed cell death (apoptosis) is now recognized as an important factor in tumour growth. Bcl-2 is an oncogene which promotes tumour progression by specifically inhibiting programmed cell death. Bcl-2 oncoprotein was measured using flow cytometry in 42 surgically excised regional lymph node metastases from patients with a median follow-up of 45 months. Fifteen patients in the study were found to have bcl-2 expression which was associated with significantly shorter survival (log-rank test, P<0.002). In addition, multivariate analysis confirmed the predictive value of bcl-2 independent of other established prognostic markers (chi(2)=7.02, P<0.01). Oncogenic control of programmed cell death is therefore important in melanoma progression and bcl-2 measurement provides a useful marker of prognosis for regional lymph node metastases.

Biomarkers, Tumor↗

The flow cytometric analysis of total p53 protein content and proliferation indices in colorectal cancer, in relation to clinical outcome.

This study was undertaken to assess the value of flow cytometric measurements of total p53 protein content and proliferation indices derived from in vivo halogenated pyrimidine labelling. Two series of colorectal cancer specimens were studied for which clinical outcome data were recorded. A series of 84 archival, ethanol-fixed, bromodeoxyuridine (BrdUrd) labelled colorectal tumours were analysed by flow cytometry for their total and cell cycle phase p53 protein content using the pAb1801 monoclonal antibody. A second series of 33 freshly obtained tumours was used for assay evaluation and for comparison with the archival material. In the archival series (n=84), the median p53-pAb1801 LI was 81.9% (range: 11.1-99.8%). In only three tumours could significant amounts of p53 protein not be detected. The median phase specific p53-pAb1801 LI in G0/G1 was 71.6%, in S was 95.5%, and in G2/M was 98.5%. In the series of fresh tumours (n=33), the median p53-pAb1801 labelling index (LI) was 94.6% (range: 17.9-99.9%). Only two tumours failed to express significant amounts of p53 protein. There was no significant difference in the generally high levels of p53 protein content between the fresh and archival series. Life-table analysis of the patients in the archival series failed to demonstrate a statistical difference in life expectancy in relation to Dukes' stage when tumours were stratified by the median total p53 labelling index. In this study, p53 content and proliferative indices measured by flow cytometry do not have independent predictive value over Dukes' grading in determining the outcome of colorectal cancer. Flow cytometry is confirmed as a practical tool for multi-parametric and cell cycle analysis of oncoprotein expression in human tumour biopsies.

Adenocarcinoma↗

Direct comparison of bromodeoxyuridine and Ki-67 labelling indices in human tumours.

Direct comparison of bromodeoxyuridine (BrdUrd) and Ki-67 labelling indices was achieved by selecting similar areas from serial sections of human tumours. Fifteen patients were selected who had been administered BrdUrd in vivo and both proliferation markers were assessed by immunohistochemistry. The data show a good correlation between both BrdUrd LI and MIB-1 LI and Tpot (calculated using the flow cytometry derived duration of S phase) and MIB-1 LI. The contribution of BrdUrd LI to growth fraction varied as a function of proliferation characteristics. In tumours with a high LI, the number of DNA synthesizing cells represented half the growth fraction, whilst in tumours with lower LI's ( < 10%) the ratio of DNA precursor labelled cells as a function of growth fraction fell to between 10% and 20%. Tpot showed a linear correlation with MIB-1/BrdUrd ratio with a slope approaching unity. It was apparent that both intra- and interpatient variation in proliferation index was greater for BrdUrd labelling than for MIB-1 expression.

Antibody Specificity↗

Conventional vs accelerated fractionation in head and neck cancer.

From October 1990 to March 1994, 90 patients entered a prospectively randomised trial in head and neck cancer. All patients had verified squamous cell carcinoma and were referred for primary radiation therapy. Tumours originated in the oral cavity in 25, oropharynx in 37, larynx in 15 and hypopharynx in 13 cases. Patients' stages were predominantely T3 and T4 (71/90) and had lymph node metastases (60/90). Seventy-nine male patients and 11 female patients, with a median age of 57 years (range 37-76 years) were treated. Patients were randomised to one of three treatment options: conventional fractionation (CF) consisting of 70 Gy in 35 fractions over 7 weeks or continuous hyperfractionated accelerated radiation therapy (Vienna-CHART) or Vienna-CHART with administration of a single dose of mitomycin C on day 5 of treatment (V-CHART + MMC). By the accelerated regimen a total dose of 55.3 Gy was given in 33 fractions within 17 consecutive days. Acute mucositis was the main toxicity recorded in those patients treated by accelerated fractionation, although the overall duration of mucosal reaction did not differ in the three treatment groups. There was no influence on local toxicity if MMC was added to radiation therapy or not. Those patients treated with additional MMC experienced a grade III/IV haematological toxicity in 4/28 cases. Complete remission (CR) was recorded in 48% following CF, 79% after Vienna-CHART (P < 0.05) and 71% after Vienna-CHART + MMC. The overall local failure rates were 73%, 59% and 42% (P = NS) for patients treated by CF, Vienna-CHART and Vienna-CHART + MMC respectively.

Adult↗

Simultaneous triple staining for hypoxia, proliferation, and DNA content in murine tumours.

Hypoxia and proliferation rate are two important biological factors influencing the outcome of radiotherapy regimes for solid tumours. Hypoxic cells are more resistant to radiation than aerobic cells and a rapidly dividing tumour may repopulate faster during treatment. Clinical trials are underway to assess the importance of both these parameters. In this article we describe a method to simultaneously measure hypoxia and proliferation using multiparameter flow cytometry. Hypoxic cells were detected using a bioreductively bound marker with an immuno-recognisable side-chain, NITP and proliferation was measured by bromodeoxyuridine (BrdUrd) incorporation. These parameters were related to cell cycle position by measuring total DNA content with 7-aminoactinomycin D. The data were analysed using single laser excitation on a bench top flow cytometer. Simultaneous measurement of the three parameters shows the presence of cells which have incorporated BrdUrd and are also hypoxic by the criterion of NITP binding. The murine SaF tumour has a relatively constant aneuploid labelling index of 24%. However, the level of aneuploid hypoxia was variable ranging from 0.8 to 40.9% with a mean value of 15.6%. Within the BrdUrd labelled population there is a range of hypoxia from 2.8 to 28.5% (mean 15.1%); this represents 0.7 to 6.6% of the total tumour population. There are approximately twice as many oxygenated cells than hypoxic cells actively in the cell cycle. In vivo tumours contain cells with S phase DNA content which do not incorporate BrdUrd. This cell population has equivalent proportions of hypoxic and oxic cells. However, there are up to 12-fold more hypoxic cells in the unlabelled S than the BrdUrd labelled population. These data show that proliferation and hypoxia can be measured simultaneously using flow cytometry and the technique may form the basis of a predictive assay for these two important biological determinants of radiotherapy outcome.

Animals↗

Pan cycle expression of proliferating cell nuclear antigen in human colorectal cancer and its proliferative correlations.

Proliferating cell nuclear antigen (PCNA) is a key cycle regulatory protein of known structure and function that also has an important role in DNA repair, its use as a marker of proliferation can be assessed directly using a thymidine analogue in suitably labelled pathological material. Following optimisation studies, a quantitative and cell cycle phase-specific analysis of PCNA expression in fresh biopsies from 50 colorectal tumours (Series A) was undertaken using dual parameter flow cytometry (FCM). For comparison, quantitative histometric studies of PCNA expression were undertaken on 50 archival bromodeoxyuridine (BrdUrd) labelled colorectal tumours (Series B). In FCM assays, PCNA-specific fluorescence was displayed throughout the cell cycle in both cells and nuclei under all preparation conditions, but to a very variable extent. The mean PCNA labelling index ranged from 38.7% to 53.0% according to the method of cell extraction used. In the 27 diploid tumours in Series A, the median PCNA LI in G0/G1 was 71.5% (range 27.0-90.6%), in S it was 10.5% (3.3-29.5%), and in G2/M it was 17.4% (5.7-43.5%). In the histometric studies of Series B tumours, the mean [S.D.] PCNA labelling index (LI) was 38.8 [9.8]%, compared with the BrdUrd (histometric) LI of 21.1 [9.0]%. The BrdUrd LI measured by FCM was 12.4 [6.5]%. PCNA-PC10 is expressed throughout the cell cycle in human colorectal tumour biopsies, which is in keeping with the range of DNA repair, synthesis, and regulatory functions that it is now recognised to perform throughout the cell cycle.

Bromodeoxyuridine↗

Proliferation measurements with flow cytometry Tpot in cancer of the uterine cervix: correlation between two laboratories and preliminary clinical results.

PURPOSE: To assess the prognostic value of the pretreatment potential doubling time (Tpot) in carcinoma of the uterine cervix, relative to other established clinical factors. METHODS AND MATERIALS: Fifty-two patients with cervical cancer were studied prospectively from March 1991 to October 1993. Pretreatment evaluation included examination under anesthesia and tumor biopsy 6 h following the intravenous administration of bromodeoxyuridine (200 mg). Tpot was determined by deriving the labeling index (LI) and S-phase synthesis time (Ts) using flow cytometry. Six patients were not evaluable and excluded. The remaining 46 patients (average age 55 years) were treated uniformly with radical radiation therapy. There were 39 squamous carcinomas and 7 adenocarcinomas. Fédération Internationale de Gynécologie et d'Obstétrique (FIGO) stages were: Ib and IIa, 12 patients; IIb, 18 patients; III and IV, 16 patients. The median external beam dose was 50 Gy (range, 45-52.8 Gy) delivered in 25 fractions. The median intracavitary dose was 40 Gy (range. 25.5-40 Gy) delivered with a single line source to a point 2 cm lateral of the midline, with a mean dose rate of 0.71 Gy/h. The median overall treatment time was 45 days (range, 34-73 days). As of July 31, 1994, 12 patients had died of disease, and the average follow-up for alive patients was 1.4 years (range, 0.5-3.3 years). RESULTS: There were 27 tumors with diploid deoxyribonucleic acid (DNA) content and 19 tumors were aneuploid. The median and mean Tpot for the 46 patients were 5.5 and 6.6 days, respectively [range, 2.0-25.6 days; coefficient of variation (CV), 74%]. For 25 patients where Tpot measurements were performed at two separate laboratories, there was a fair correlation (r = 0.74), but systematic differences were detected suggesting that the lack of agreement was not simply due to intratumoral variation. To date, 30 patients remained disease free, while 8 patients had pelvic failure and 9 patients developed distant metastases as the first failure site (1 patient developed both at the same time). In univariate analysis, the only significant prognostic factor for disease-free survival was tumor size (p = 0.004). A short Tpot (or high LI) and long overall treatment time (OTT) were weakly associated with poorer disease-free survival, although not statistically significant (1/Tpot, p = 0.14; LI, p = 0.23; OTT, p = 0.04). Age, FIGO stage, hemoglobin level, S-phase fraction, DNA ploidy, and Ts were not associated with disease-free survival. Multiple regression analysis was not performed because of the relatively small number of patients and short follow-up. CONCLUSIONS: Tpot values determined with current techniques by different laboratories cannot be used interchangeably for the purpose of therapy decisions. Vigorous quality assurance and standardization of the laboratory procedures and analysis methods are important to reduce interlaboratory variation. In this uniformly treated group of patients with cancer of the uterine cervix, traditional clinical prognostic factors remain the most important. Preliminary data suggest that the flow cytometry-determined Tpot and labeling index predict for disease-free survival, although a larger number of patients with longer follow-up is required to assess the true prognostic significance of these assays and to determine if their effect is independent of other clinical factors.

Adenocarcinoma↗

Radiation-induced changes in glomerular and tubular cell kinetics and morphology following irradiation of a single kidney in the pig.

PURPOSE: Radiation-induced changes in glomerular and tubular cell kinetics and morphology following irradiation of a single pig kidney were assessed. METHODS AND MATERIALS: The right kidney of 13 adult female Large White pigs was irradiated with a single dose of 9.8 Gy gamma rays. Animals were serially killed between 2 and 24 weeks postirradiation (PI); 1 h prior to postmortem each pig received 500 mg bromodeoxyuridine (BrdUrd). At postmortem, both kidneys were removed and tissue taken to prepare cell suspensions. The labeling index (LI) of these suspensions was measured using flow cytometry; in vivo BrdUrd incorporation in glomerular and tubular cells was determined immunohistochemically. The kidneys were also assessed histologically. RESULTS: Irradiation of the right kidney alone resulted in a significant increase in renal cell LI in both the irradiated and the contralateral unirradiated kidney within 2 weeks of irradiation; peak values of 1.57 +/- 0.32% and 1.04 +/- 0.13%, respectively, were seen 4 weeks PI, significantly greater (P < 0.001) than the preirradiation value of 0.18 +/- 0.01%. The LI values then declined with time, but remained greater than those seen prior to irradiation. A similar pattern of response was determined from counts of labeled glomerular and tubular cells identified immunohistochemically. The increase in labeled glomerular cells was seen 2 weeks PI, whereas that for the tubular cells did not occur until 4 weeks PI. The irradiated kidney exhibited diffuse, progressive glomerular alterations. In contrast, tubular damage was focal; the irradiated kidney also exhibited a prominent vasculopathy, involving arteriolar and peripheral interlobular artery thickening. The contralateral unirradiated kidney appeared unchanged. CONCLUSION: These findings confirm the hypothesis that the morphologic and kinetic responses observed after irradiation of a single kidney are similar to those observed after irradiation of both kidneys. Renal irradiation results in significant alterations in glomerular and tubular cell proliferation and morphology within 2-4 weeks of irradiation; glomerular changes appear predominant.

Animals↗

Chromosome abnormalities and p53 expression in a small cell carcinoma of the bladder.

Chromosome studies on a highly malignant tumor, a small cell carcinoma of the bladder (the first to be studied cytogenetically), showed a hypertriploid mainline and a hypertetraploid minor line. Extensive chromosomal rearrangements were present in both lines, some rearranged chromosomes being seen in only one of the lines, while others, derived from chromosomes 6, 9, 11, 13, and 18, were seen in both. Although different giant chromosomes were present in the two lines, they shared a possibly significant common feature: multiple copies of 2q. DNA flow cytometry confirmed that the tumor had a hypertriploid main mode and showed that dysplastic surface epithelium present in the histologic material also had a hypertriploid DNA index. p53 expression in the tumor was demonstrated by flow cytometry.

Carcinoma, Small Cell↗

Studies with bromodeoxyuridine in head and neck cancer and accelerated radiotherapy.

Using bromodeoxyuridine (BrdUrd), tumour cell proliferation was assessed, by flow cytometric (FCM) and immunohistochemical methods, in patients treated by the CHART regime of radiotherapy. Of 115 cases studied, data were complete in 90 using both methods. No cell kinetic-related parameter predicted the outcome of patients treated by CHART, in keeping with the view that, with this regime, cellular proliferation had been minimised as a cause of failure. Histological evaluation of the labelling index (LI) revealed a trend for higher LI is in diploid tumours (16.2%) than in aneuploid (13.8%), contrasting to that found by FCM (5.0 and 9.3% respectively). When the Tpot was calculated using a combination of histology LI and FCM TS, diploid tumours showed more rapid proliferation (Tpot 1.8 days) than aneuploid tumours (Tpot 3.2 days); this finding was significant (p < 0.02). A novel parameter, termed proliferation pattern, unique to these studies with BrdUrd in vivo, was assessed. Both proliferation pattern and histological grading had predictive power to discriminate the outcome in univariate analysis (p = < 0.031 and 0.037, respectively). In a Cox multivariate analysis, proliferation pattern was the more important predictor. The studies reported highlight the extra information that can be gained from combining immunohistochemistry with flow cytometry to study the cellular proliferation of human tumours.

Bromodeoxyuridine↗

p53 status of head and neck cancer: relation to biological characteristics and outcome of radiotherapy.

p53 status was investigated in 99 patients with squamous cell carcinoma of the head and neck region uniformly treated with accelerated radiotherapy and in whom tumour cell proliferation and DNA aneuploidy were assessed using bromodeoxyuridine (BrdUrd) incorporation and flow cytometry (FCM). Seventy-six percent of tumours were immunohistochemically positive for p53 protein, but heterogeneity was noticed both in the percentage of cells positive for p53 and in their level of expression. However, tumours which were either essentially all positive or all negative or showed sporadic positivity for p53 protein showed no differences in their level of aneuploidy, proliferation rate, tissue organisation or outcome with radiotherapy. There was a trend for those p53-positive tumours with the strongest expression to have more DNA aneuploidy and deregulation of proliferation organisation than weaker expressors; but there were no differences in proliferation rate or outcome of radiotherapy. These studies suggest that p53 protein stabilisation as assessed by immunohistochemistry does not have any major relationship with the biological characteristics and outcome of squamous cell cancer treated by accelerated radiotherapy.

Aneuploidy↗

Development of bioreductive markers for tumour hypoxia.

Hypoxic cells in tumours can be identified by exposing them to an immunologically identifiable 2-nitroimidazole (NITP) with a theophylline substituent which becomes bioreductively metabolised and binds to cellular macromolecules in the absence of oxygen. A range of monoclonal and polyclonal antibodies raised against theophylline or caffeine can identify cells containing bound adducts of NITP, in some cases with higher specificity than the standard product used. An alternative approach utilizes the very high specificity of FITC-avidin as a reagent to detect metabolic binding of a 2-nitroimidazole with a biotinylated side-chain (NIB), with the advantage of a single-step staining protocol. Both proliferating and hypoxic cell populations within tumours can be identified by simultaneous staining for incorporation of NITP and BrdUrd and this has shown that some cells incorporate both markers, suggesting that there is some overlap between the proliferating and hypoxic cell compartments.

Animals↗

A test to estimate VO2max in females using aerobic dance, heart rate, BMI, and age.

OBJECTIVE: The purpose of this investigation was to develop a sub-maximal exercise test for estimating VO2max utilizing aerobic dance. EXPERIMENTAL DESIGN: One hundred females between the ages 18 to 40 yr served as the subjects for test validation. The subjects completed a treadmill test to determine VO2max and were assessed for heart rate (HR) response to a bout of aerobic dance. The data associated with responses to treadmill exercise and the aerobic dance test, in conjunction with descriptive variables (e.g., age, BMI) were utilized in the validation of the multiple regression model. MEASURES: Reliability was determined by correlation and paired "t"-tests of the aerobic dance routine test and retest trials. The construction of the multiple regression equation, via forward entry analysis, and the cross-validation of the regression equation were completed to ensure the validity and reliability of the protocol in accurately estimating VO2max. RESULTS: Test, retest reliability for the dance-exercise routine was demonstrated (r = 0.98). Moreover, no significant differences were shown between the HR responses for the test and retest trials. The multiple regression analysis yielded a three variable multiple prediction equation for estimating VO2max (R = 0.84; SEE, 5.5 ml.kg-1.min-1). The three variables were the HR response to four min of aerobic dance (HR4), body mass index (BMI), and age (years). Cross-validation of the aerobic dance test was determined with a second group of 50 female subjects (R = 0.83; SEE, 5.5 ml.kg-1.min-1). Additionally, the application of the validation group regression equation to the cross-validation group yielded a comparable R of 0.82. Comparison of the predicted values for VO2max from both equations also yielded a highly significant invariance coefficient of 0.96. Finally, the results of "t"-tests between the observed and predicted mean values for VO2max revealed no significant difference (p > 0.05). Therefore, the final prediction equation, based on collapsing the means associated with the validation and cross validation samples (n = 150), was: VO2max (ml.kg-1.min-1) = 130.18-(0.38 * HR4)-(0.81 * BMI)-(0.27 * Age); R = 0.84, SEE = 5.5 ml.kg-1.min-1. CONCLUSIONS: These results indicate that a four minute aerobic dance test provides a valid and reliable sub-maximal protocol for estimating VO2max and providing an index of aerobic fitness in apparently healthy 18 to 40 yr old females.

Adolescent↗

Correlation between p53 gene expression and tumor-cell proliferation in oropharyngeal cancer.

The aim of our study was to analyze the correlation between tumor-cell kinetics measured in vivo and p53 gene expression in a series of 49 oropharyngeal cancers. The duration of the S phase (TS), the labelling index (LI) and the potential doubling time (Tpot) were obtained by flow-cytometry measurements of a tumor biopsy obtained after i.v. injection of 200 mg bromodeoxyuridine into patients. An adjacent section of the same samples was studied by immunohistochemistry for the detection of p53 protein. Over-expression of the p53 protein, as defined by strong p53 immunostaining of tumor nuclei, was found in 23/49 of the samples. Aneuploid tumors showed a higher LI, a shorter Tpot and a higher proportion of p53 gene over-expression. A significant correlation was seen between p53 over-expression and short Tpots. Indeed 87.5% of the tumors with a very short Tpot (< or = 3 days) had p53 over-expression, as compared with 27% of tumors with a longer Tpot (> 3 days). Our data strongly suggest that over-expression of the p53 gene is associated with rapid tumor-cell proliferation in this type of cancer.

Aged↗

Radiation-induced cell cycle delay measured in two mouse tumors in vivo using bromodeoxyuridine.

The magnitude of the delay of cells in the phases of the cell cycle after irradiation may be related to the radioresponsiveness of tumor cell populations. In this study we have quantified division delay in two mouse tumors in vivo after single and fractionated doses of X rays and single doses of neutrons. The incorporation of bromodeoxyuridine and flow cytometry provided a sensitive and quantitative method to detect cell cycle perturbations after radiation treatment. The more rapidly growing SAF tumor showed less G2-phase delay per gray than a more slowly proliferating tumor, the Rh (0.9 vs 1.8 h). In addition, the SAF tumor failed to show any G1/S-phase delay while the Rh tumor experienced a longer G1-phase delay than that measured for G2 phase (3.1 vs 1.8 h). There was a trend in both tumors for lower doses to be more effective in producing cell cycle delays. Neutrons caused longer G2-phase delays on a unit dose basis, 2.5 and 5.4 h for the SAF and Rh tumors, respectively. The RBE for neutrons for division delay was found to be 2.9 and 2.8 for the SAF and Rh tumors, while the RBE for growth delay was 3.4 and 3.5. Fractionation of the X-ray dose caused a reduction in division delay at higher total doses (10 or 12 Gy) but was without effect at the lower dose studied (6 Gy). These studies show the feasibility of measuring cell cycle delays in vivo, and future developments are suggested for a possible predictive test in patients receiving radiotherapy.

Animals↗