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RAD001 (everolimus) inhibits tumour growth in xenograft models of human hepatocellular carcinoma.

Huynh H, Chow KH, Soo KC, Toh HC, Choo SP, Foo KF, Poon D, Ngo VC, Tran E - J. Cell. Mol. Med. (2008)

Bottom Line: Hepatocellular carcinoma (HCC) is the fifth most common malignancy worldwide and highly resistant to available chemotherapies.RAD001-induced growth suppression was associated with inactivation of downstream targets of mTOR, reduction in VEGF expression and microvessel density, inhibition of cell proliferation, up-regulation of p27(Kip1) and down-regulation of p21(Cip1/Waf1), Cdk-6, Cdk-2, Cdk-4, cdc-25C, cyclin B1 and c-Myc.Our study provides a strong rationale for clinical investigation of mTOR inhibitor RAD001 in patients with HCC.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Molecular Endocrinology, Division of Cellular and Molecular Research, Humphrey Oei Institute of Cancer Research, National Cancer Centre, Singapore. cmrhth@nccs.com.sg

ABSTRACT
Hepatocellular carcinoma (HCC) is the fifth most common malignancy worldwide and highly resistant to available chemotherapies. Mammalian target of rapamycin (mTOR) functions to regulate protein translation, angiogenesis and cell cycle progression in many cancers including HCC. In the present study, subcutaneous patient-derived HCC xenografts were used to study the effects of an mTOR inhibitor, RAD001 (everolimus), on tumour growth, apoptosis and angiogenesis. We report that oral administration of RAD001 to mice bearing patient-derived HCC xenografts resulted in a dose-dependent inhibition of tumour growth. RAD001-induced growth suppression was associated with inactivation of downstream targets of mTOR, reduction in VEGF expression and microvessel density, inhibition of cell proliferation, up-regulation of p27(Kip1) and down-regulation of p21(Cip1/Waf1), Cdk-6, Cdk-2, Cdk-4, cdc-25C, cyclin B1 and c-Myc. Our data indicate that the mTOR pathway plays an important role in angiogenesis, cell cycle progression and proliferation of liver cancer cells. Our study provides a strong rationale for clinical investigation of mTOR inhibitor RAD001 in patients with HCC.

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Effects of RAD001 on growth inhibition of patient-derived HCC xenografts. Indicated xenograft lines were s.c. implanted in SCID mice as described in Materials and Methods. For the dose-response experiment, mice bearing xenografts were treated with vehicle or five doses (1, 1.5, 2, 2.5 and 5 mg) of RAD001 per kg body weight daily for 18 days as described in Materials and Methods. To study the effects of RAD001 on growth rate, mice bearing 26-1004 tumours were treated with vehicle or three doses of RAD001 (1, 2.5 and 5) mg/kg for 18 days. For 5-1318 and 30-1004 xenografts, only 2.5 mg dose was used. Each treatment arm involved 14 independent tumour-bearing mice representing the same xenograft line. Tumour growth was measured and calculated as described in Materials and Methods. Mean volume ± S.E. at a given time for vehicle- or RAD001-treated tumours is shown. The differences seen were statistically significant (P < 0.01). Experiments were repeated three times with similar results.
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fig02: Effects of RAD001 on growth inhibition of patient-derived HCC xenografts. Indicated xenograft lines were s.c. implanted in SCID mice as described in Materials and Methods. For the dose-response experiment, mice bearing xenografts were treated with vehicle or five doses (1, 1.5, 2, 2.5 and 5 mg) of RAD001 per kg body weight daily for 18 days as described in Materials and Methods. To study the effects of RAD001 on growth rate, mice bearing 26-1004 tumours were treated with vehicle or three doses of RAD001 (1, 2.5 and 5) mg/kg for 18 days. For 5-1318 and 30-1004 xenografts, only 2.5 mg dose was used. Each treatment arm involved 14 independent tumour-bearing mice representing the same xenograft line. Tumour growth was measured and calculated as described in Materials and Methods. Mean volume ± S.E. at a given time for vehicle- or RAD001-treated tumours is shown. The differences seen were statistically significant (P < 0.01). Experiments were repeated three times with similar results.

Mentions: We next examined anti-tumour activity of RAD001 in these xenografts. As shown in Figure 2A, RAD001 inhibited the growth of HCC xenografts in a dose-dependent manner (P < 0.01). The xenograft line 26-1004, which has a 16 base pair deletion in exon 8 of the PTEN gene, had the greatest sensitivity to RAD001 as measured by percentage of growth inhibition (Fig. 2A). No overt toxicity of RAD001 was observed during the course of treatment as defined by weight loss, unkempt appearance or mortality, and behaviour. Since the dose of 2.5 mg/kg daily gave maximal growth inhibition, we selected this dose for our subsequent studies. Reduction of tumour growth rates rather than producing regressions was noticed in all xenografts studied (Fig. 2B–D). The growth suppression was seen approximately 1 week after treatment. RAD001, when given at a dose of 2.5 mg/kg daily, significantly inhibited the growth of all four xenograft lines (P < 001, Fig. 3). The T/C ratio, where T and C are the median weight (mg) of RAD001-treated and vehicle-treated tumours at day 18 during the treatment, respectively, for 26-1004, 2-1318, 5-1318 and 30-1004 xenografts was 0.11, 0.19, 0.14 and 0.48, respectively. Since the ratios of less than 0.42 were observed in three of four lines studied, it was considered an active response (Drug Evaluation Branch of the Division of Cancer Treatment, NCI criteria).


RAD001 (everolimus) inhibits tumour growth in xenograft models of human hepatocellular carcinoma.

Huynh H, Chow KH, Soo KC, Toh HC, Choo SP, Foo KF, Poon D, Ngo VC, Tran E - J. Cell. Mol. Med. (2008)

Effects of RAD001 on growth inhibition of patient-derived HCC xenografts. Indicated xenograft lines were s.c. implanted in SCID mice as described in Materials and Methods. For the dose-response experiment, mice bearing xenografts were treated with vehicle or five doses (1, 1.5, 2, 2.5 and 5 mg) of RAD001 per kg body weight daily for 18 days as described in Materials and Methods. To study the effects of RAD001 on growth rate, mice bearing 26-1004 tumours were treated with vehicle or three doses of RAD001 (1, 2.5 and 5) mg/kg for 18 days. For 5-1318 and 30-1004 xenografts, only 2.5 mg dose was used. Each treatment arm involved 14 independent tumour-bearing mice representing the same xenograft line. Tumour growth was measured and calculated as described in Materials and Methods. Mean volume ± S.E. at a given time for vehicle- or RAD001-treated tumours is shown. The differences seen were statistically significant (P < 0.01). Experiments were repeated three times with similar results.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4496150&req=5

fig02: Effects of RAD001 on growth inhibition of patient-derived HCC xenografts. Indicated xenograft lines were s.c. implanted in SCID mice as described in Materials and Methods. For the dose-response experiment, mice bearing xenografts were treated with vehicle or five doses (1, 1.5, 2, 2.5 and 5 mg) of RAD001 per kg body weight daily for 18 days as described in Materials and Methods. To study the effects of RAD001 on growth rate, mice bearing 26-1004 tumours were treated with vehicle or three doses of RAD001 (1, 2.5 and 5) mg/kg for 18 days. For 5-1318 and 30-1004 xenografts, only 2.5 mg dose was used. Each treatment arm involved 14 independent tumour-bearing mice representing the same xenograft line. Tumour growth was measured and calculated as described in Materials and Methods. Mean volume ± S.E. at a given time for vehicle- or RAD001-treated tumours is shown. The differences seen were statistically significant (P < 0.01). Experiments were repeated three times with similar results.
Mentions: We next examined anti-tumour activity of RAD001 in these xenografts. As shown in Figure 2A, RAD001 inhibited the growth of HCC xenografts in a dose-dependent manner (P < 0.01). The xenograft line 26-1004, which has a 16 base pair deletion in exon 8 of the PTEN gene, had the greatest sensitivity to RAD001 as measured by percentage of growth inhibition (Fig. 2A). No overt toxicity of RAD001 was observed during the course of treatment as defined by weight loss, unkempt appearance or mortality, and behaviour. Since the dose of 2.5 mg/kg daily gave maximal growth inhibition, we selected this dose for our subsequent studies. Reduction of tumour growth rates rather than producing regressions was noticed in all xenografts studied (Fig. 2B–D). The growth suppression was seen approximately 1 week after treatment. RAD001, when given at a dose of 2.5 mg/kg daily, significantly inhibited the growth of all four xenograft lines (P < 001, Fig. 3). The T/C ratio, where T and C are the median weight (mg) of RAD001-treated and vehicle-treated tumours at day 18 during the treatment, respectively, for 26-1004, 2-1318, 5-1318 and 30-1004 xenografts was 0.11, 0.19, 0.14 and 0.48, respectively. Since the ratios of less than 0.42 were observed in three of four lines studied, it was considered an active response (Drug Evaluation Branch of the Division of Cancer Treatment, NCI criteria).

Bottom Line: Hepatocellular carcinoma (HCC) is the fifth most common malignancy worldwide and highly resistant to available chemotherapies.RAD001-induced growth suppression was associated with inactivation of downstream targets of mTOR, reduction in VEGF expression and microvessel density, inhibition of cell proliferation, up-regulation of p27(Kip1) and down-regulation of p21(Cip1/Waf1), Cdk-6, Cdk-2, Cdk-4, cdc-25C, cyclin B1 and c-Myc.Our study provides a strong rationale for clinical investigation of mTOR inhibitor RAD001 in patients with HCC.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Molecular Endocrinology, Division of Cellular and Molecular Research, Humphrey Oei Institute of Cancer Research, National Cancer Centre, Singapore. cmrhth@nccs.com.sg

ABSTRACT
Hepatocellular carcinoma (HCC) is the fifth most common malignancy worldwide and highly resistant to available chemotherapies. Mammalian target of rapamycin (mTOR) functions to regulate protein translation, angiogenesis and cell cycle progression in many cancers including HCC. In the present study, subcutaneous patient-derived HCC xenografts were used to study the effects of an mTOR inhibitor, RAD001 (everolimus), on tumour growth, apoptosis and angiogenesis. We report that oral administration of RAD001 to mice bearing patient-derived HCC xenografts resulted in a dose-dependent inhibition of tumour growth. RAD001-induced growth suppression was associated with inactivation of downstream targets of mTOR, reduction in VEGF expression and microvessel density, inhibition of cell proliferation, up-regulation of p27(Kip1) and down-regulation of p21(Cip1/Waf1), Cdk-6, Cdk-2, Cdk-4, cdc-25C, cyclin B1 and c-Myc. Our data indicate that the mTOR pathway plays an important role in angiogenesis, cell cycle progression and proliferation of liver cancer cells. Our study provides a strong rationale for clinical investigation of mTOR inhibitor RAD001 in patients with HCC.

Show MeSH
Related in: MedlinePlus