50 mg/kg100 mg/kg

MCF-7/HER2-18?Ki67 (%)77.7 (74.4C81.1)69.6** (63.4C74.8)65.5**** (62.0C70.1)?TUNEL (%)1.2 (0.9C1.5)1.5* (1.2C1.6)1.6** (1.4C1.9)?CTI61.6 (55.5C79.5).48.7*** (41.6C57.4)38.0**** (30.1C43.3)SKOV3?Ki67 (%)46.6 (32.0C63.8)34.1** (20.3C57.2)40.1 (26.6C48.8)?TUNEL (%)0.35 (0.1C0.8)0.49 (0.1C1.0)0.48 (0.1C1.4)?CTI125.4 (87.1C188.9).67.5**(45.6C96.1)81.0* (38.6C110.2)MDA-MB231?Ki67 (%)61.7 (54.1C74.6)69.3 (56.1C78.0)72.4* (43.8C83.1)?TUNEL (%)0.55 (0.3C0.9)0.45 (0.3C0.6)0.31* (0.1C0.4)?CTI106.0 (85.8C191.0).180.1 (115.9C205.4)180.8 (90.5C227.7) Open in a separate window Effect of R115777 on proliferation (Ki67), apoptosis (TdT-mediated dUTP-fluorescence nick end labelling [TUNEL]) and cell turnover index (CTI) in cell line tumours grown in athymic nude mice. Values are expressed as median values (interquartile range). *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. Growth of SKOV3 tumours was inhibited by R115777 50 mg/kg (n = 30) and 100 mg/kg (n = 14) by 60.1% (32.3C83.9%; P = 0.04) and 20.4% (-65.7 to +38.3%; P = 0.4), respectively, as compared with that in control tumours (n = 14; Figure ?Figure1b1b and Table ?Table1).1). The proliferation index was reduced treated tumours; for R115777 50 mg/kg it was 34.1% (26.5C44.4%; P = 0.009) and for R115777 100 mg/kg it was 40.1% (33.1C44.0%; P = 0.08) as compared with 46.6% in the control tumours (37.6C55.0%). The apoptotic index did not differ between treated tumours; for R115777 50 mg/kg it was 0.49% (0.4C0.7%; P = 0.11), for R115777 100 mg/kg it was 0.48% (0.4C0.8%; P = 0.18) and it was 0.35% in the control tumours (0.3C0.5%). The CTIs for tumours treated with R115777 50 mg/kg and 100 mg/kg were 67.5 (45.6C96.1; P = 0.004) and 81.0 (38.6C110.2; P = 0.05), respectively; these values were statistically significantly reduced as compared with the CTI of 125.4 in controls (87.1C188.9; Table ?Table22). Growth of k-ras mutated MDA-MB231 tumours was not inhibited by R115777 50 mg/kg (n = 25) and 100 mg/kg (n = 15). Instead, the growth in the treated tumours was increased by 68.8% (13.8C284.1%; P = 0.08) and 91.2% (2.8C328.8%; P = 0.09), respectively, relative to control tumours (n = 16; Figure ?Figure1c1c and Table ?Table1).1). The proliferation index was higher, although not statistically significantly higher, in the treated tumours; for R115777 50 mg/kg it was 68.2% (63.3C71.5%; P = 0.24) and for R115777 100 mg/kg it was 72.4% (66.4C78.4%; P = 0.06) as compared with 57.9% in the control tumours (54.1C71.4%). The apoptotic index was similar in treated and control tumours; for R115777 50 mg/kg it was 0.4% (0.3C0.6%; P =.In one study [38] two out of 25 DCIS lesions over-expressed wild-type k-ras, and in another study [39] four out of five DCIS individuals expressed detectable plasma levels of wild-type ras, but no ras mutations were found in these studies. We have shown the FTI was effective in some tumours and not in others. were two sided. Statistical analysis was performed using STATA software (STATA Corporation, College Train station, TX, USA). Results Cell growth IC50 for R115777 assorted a hundred-fold, from 39 nmol/l and 46 nmol/l for SK-BR3 and MCF-7 to 2.7 mol/l and 5.9 mol/l for SKOV3 and MDA-MB231 cell lines, respectively, when cultivated in vitro (Table ?(Table1).1). The cell collection with the mutated k-ras, namely MDA-MB231, had the highest IC50, whereas in the cell lines with wild-type ras the drug was effective at lower doses. In mice, growth of MCF-7/HER2-18 tumours was inhibited by R115777 50 mg/kg (n = 19) and 100 mg/kg (n = 11) by 80.8% (interquartile range 56.4C99.0%; P = 0.001) and 95.9% (68.2C110.1%; P = 0.02), respectively, compared with control tumours (n = 22; Figure ?Figure1a1a and Table ?Table1).1). The proliferation index was lower in the treated tumours; for R115777 50 mg/kg it was 69.6% (63.4C74.8%; P = 0.003) and for R115777 100 mg/kg it was 65.5% (62.0C70.1%; P < 0.0001) as compared with 77.7% for the control tumours (74.4C81.1%; Table ?Table2).2). The apoptotic index was higher in the treated tumours; for R115777 50 mg/kg it was 1.5% (1.2C1.6%; P = 0.04) and for R115777 100 mg/kg it was 1.6% (1.4C1.9%; P = 0.003) as compared with 1.2% in the control tumours (0.9C1.5%). The CTIs for tumours treated with R115777 50 mg/kg and 100 mg/kg were 48.7 (41.6C57.4; P = 0.0009) and 38.0 (30.1C43.3; P < 0.0001), respectively, and these values were statistically significantly reduced as compared with the CTI of 61.6 in controls (55.5C79.5; Table ?Table22). Open in a separate window Figure 1 Tumour growth inhibition. (a) Tumour growth inhibition in MCF-7/HER2-18 tumours grown in athymic mice treated with R115777 50 mg/kg or 100 mg/kg. Median tumour volume in treated relative to control animals is given in cubic millimetres. (b) Tumour growth inhibition by R115777 in SKOV3 tumours. (c) Tumour growth inhibition by R115777 in MDA-MB231 tumours. Table 2 Apoptosis and proliferation in cell tumour experiments Cell lineControlTreated R115777

50 mg/kg100 mg/kg

MCF-7/HER2-18?Ki67 (%)77.7 (74.4C81.1)69.6** (63.4C74.8)65.5**** (62.0C70.1)?TUNEL (%)1.2 (0.9C1.5)1.5* (1.2C1.6)1.6** (1.4C1.9)?CTI61.6 (55.5C79.5).48.7*** (41.6C57.4)38.0**** (30.1C43.3)SKOV3?Ki67 (%)46.6 (32.0C63.8)34.1** (20.3C57.2)40.1 (26.6C48.8)?TUNEL (%)0.35 (0.1C0.8)0.49 (0.1C1.0)0.48 (0.1C1.4)?CTI125.4 (87.1C188.9).67.5**(45.6C96.1)81.0* (38.6C110.2)MDA-MB231?Ki67 (%)61.7 (54.1C74.6)69.3 (56.1C78.0)72.4* (43.8C83.1)?TUNEL (%)0.55 (0.3C0.9)0.45 (0.3C0.6)0.31* (0.1C0.4)?CTI106.0 (85.8C191.0).180.1 (115.9C205.4)180.8 (90.5C227.7) Open in a separate window Effect of R115777 on proliferation (Ki67), apoptosis (TdT-mediated dUTP-fluorescence nick end labelling [TUNEL]) and cell turnover index (CTI) in cell line tumours grown in athymic nude mice. Values are expressed as median values (interquartile range). *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. Growth of SKOV3 tumours was inhibited by R115777 50 mg/kg (n = 30) and 100 mg/kg (n = 14) by 60.1% (32.3C83.9%; P = 0.04) and 20.4% (-65.7 to +38.3%; P = 0.4), respectively, as compared with that in control tumours (n = 14; Figure ?Figure1b1b and Table ?Table1).1). The proliferation index was lower in treated tumours; for R115777 50 mg/kg it was 34.1% (26.5C44.4%; P = 0.009) and for R115777 100 mg/kg it was 40.1% (33.1C44.0%; P = 0.08) as compared with 46.6% in the control tumours (37.6C55.0%). The apoptotic index did not differ between treated tumours; for R115777 50 mg/kg it was 0.49% (0.4C0.7%; P = 0.11), for R115777 100 mg/kg it was 0.48% (0.4C0.8%; P = 0.18) and it was 0.35% in the control tumours (0.3C0.5%). The CTIs for tumours treated with R115777 50 mg/kg and 100 mg/kg were 67.5 (45.6C96.1; P = 0.004) and 81.0 (38.6C110.2; P = 0.05), respectively; these values were statistically significantly reduced as compared with the CTI of 125.4 in controls (87.1C188.9; Table ?Table22). Growth of k-ras mutated MDA-MB231 tumours was not inhibited by R115777 50 mg/kg (n = 25) and 100 mg/kg (n = 15). Instead, the growth in the treated tumours was increased by 68.8% (13.8C284.1%; P = 0.08) and 91.2% (2.8C328.8%; P = 0.09), respectively, relative to control tumours (n = 16; Figure ?Figure1c1c and Table ?Table1).1). The proliferation index was higher, although not statistically significantly higher, in the treated tumours; for R115777 50 mg/kg it was 68.2% (63.3C71.5%; P = 0.24) and for.The CTIs for tumours treated with R115777 50 mg/kg and 100 mg/kg were 180.1 (115.9C205.4; P = 0.32) and 180.8 (90.5C227.7; P = 0.37), respectively, and the CTI for controls was 106.0 (85.8C191.0; Table ?Table22). In the xenograft experiments no difference in body weight was seen between R115777 and controls; neither was there any difference in the number of deaths seen between treated and control animals. Ductal carcinoma in situ Tissue from 14 women with widespread DCIS was implanted. control mice. Throughout the analysis, ‘mouse’ was taken as the unit of analysis and only data from mice with data from both day 14 and day 21 or day 28 were included, because we looked at change over time in terms of proliferation, apoptosis and CTIs. Data are presented as the geometric means with their 95% confidence intervals. The significance level was set at 5% and all tests were two sided. Statistical analysis was performed using STATA software (STATA Corporation, College Station, TX, USA). Results Cell growth IC50 for R115777 varied a hundred-fold, from 39 nmol/l and 46 nmol/l for SK-BR3 and MCF-7 to 2.7 mol/l and 5.9 mol/l for SKOV3 and MDA-MB231 cell lines, respectively, when produced in vitro (Table ?(Table1).1). The cell line with the mutated k-ras, namely MDA-MB231, had the highest IC50, whereas in the cell lines with wild-type ras the drug was effective at lower doses. In mice, growth of MCF-7/HER2-18 tumours was inhibited by R115777 50 mg/kg (n = 19) and 100 mg/kg (n = 11) by 80.8% (interquartile range 56.4C99.0%; P = 0.001) and 95.9% (68.2C110.1%; P = 0.02), respectively, compared with control tumours (n = 22; Figure ?Figure1a1a and Table ?Table1).1). The proliferation index was lower in the treated tumours; for R115777 50 mg/kg it was 69.6% (63.4C74.8%; P = 0.003) and for R115777 100 mg/kg it was 65.5% (62.0C70.1%; P < 0.0001) as compared with 77.7% for the control tumours (74.4C81.1%; Table ?Table2).2). The apoptotic index was higher in the treated tumours; for R115777 50 mg/kg it was 1.5% (1.2C1.6%; P = 0.04) and for R115777 100 mg/kg it was 1.6% (1.4C1.9%; P = 0.003) as compared with 1.2% in the control tumours (0.9C1.5%). The CTIs for tumours treated with R115777 50 mg/kg and 100 mg/kg were 48.7 (41.6C57.4; P = 0.0009) and 38.0 (30.1C43.3; P < 0.0001), respectively, and these values were statistically significantly reduced as compared with the CTI of 61.6 in controls (55.5C79.5; Table ?Table22). Open in a separate window Figure 1 Tumour growth inhibition. (a) Tumour growth inhibition in MCF-7/HER2-18 tumours grown in athymic mice treated with R115777 50 mg/kg or 100 mg/kg. Median tumour volume in treated relative to control animals is given in cubic millimetres. (b) Tumour growth inhibition by R115777 in SKOV3 tumours. (c) Tumour NSC 3852 growth inhibition by R115777 in MDA-MB231 tumours. Table 2 Apoptosis and proliferation in cell tumour experiments

50 mg/kg100 mg/kg

MCF-7/HER2-18?Ki67 (%)77.7 (74.4C81.1)69.6** (63.4C74.8)65.5**** (62.0C70.1)?TUNEL (%)1.2 (0.9C1.5)1.5* (1.2C1.6)1.6** (1.4C1.9)?CTI61.6 (55.5C79.5).48.7*** (41.6C57.4)38.0**** (30.1C43.3)SKOV3?Ki67 (%)46.6 (32.0C63.8)34.1** (20.3C57.2)40.1 (26.6C48.8)?TUNEL (%)0.35 (0.1C0.8)0.49 (0.1C1.0)0.48 (0.1C1.4)?CTI125.4 (87.1C188.9).67.5**(45.6C96.1)81.0* (38.6C110.2)MDA-MB231?Ki67 (%)61.7 (54.1C74.6)69.3 (56.1C78.0)72.4* (43.8C83.1)?TUNEL (%)0.55 (0.3C0.9)0.45 (0.3C0.6)0.31* (0.1C0.4)?CTI106.0 (85.8C191.0).180.1 (115.9C205.4)180.8 (90.5C227.7) Open in a separate window Effect of R115777 on proliferation (Ki67), apoptosis NSC 3852 (TdT-mediated dUTP-fluorescence nick end labelling [TUNEL]) and cell turnover index (CTI) in cell line tumours grown in athymic nude mice. Values are expressed as median values (interquartile range). *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. Growth of SKOV3 tumours was inhibited by R115777 50 mg/kg (n = 30) and 100 mg/kg (n = 14) by 60.1% (32.3C83.9%; P = 0.04) and 20.4% (-65.7 to +38.3%; P = 0.4), respectively, as compared with that in control tumours (n = 14; Figure ?Figure1b1b and Table ?Table1).1). The proliferation index was lower in treated tumours; for R115777 50 mg/kg it was 34.1% (26.5C44.4%; P = 0.009) and for R115777 100 mg/kg it was 40.1% (33.1C44.0%; P = 0.08) as compared with 46.6% in the control tumours (37.6C55.0%). The apoptotic index did not differ between treated tumours; for R115777 50 mg/kg it was 0.49% (0.4C0.7%; P = 0.11), for R115777 100 mg/kg it was 0.48% (0.4C0.8%; P = 0.18) and it was 0.35% in the control tumours NSC 3852 (0.3C0.5%). The CTIs for tumours treated with R115777 50 mg/kg and 100 mg/kg were 67.5 (45.6C96.1;.