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Inhibition of angiogenic and non-angiogenic targets by sorafenib in renal cell carcinoma (RCC) in a RCC xenograft model.

Yuen JS, Sim MY, Siml HG, Chong TW, Lau WK, Cheng CW, Huynh H - Br. J. Cancer (2011)

Bottom Line: Sorafenib-induced growth suppression was associated with not only inhibition of angiogenic targets p-PDGFR-β, p-VEGFR-2, and their downstream signalling pathways p-Akt and p-ERK, cell cycle, and anti-apoptotic proteins that include cyclin D1, cyclin B1, and survivin but also upregulation of proapoptotic Bim.This study has shed light on the molecular mechanisms of sorafenib in RCC.Inhibition of non-angiogenic molecules by sorafenib could contribute in part to its anti-tumour activities observed in vivo, in addition to its anti-angiogenic effects.

View Article: PubMed Central - PubMed

Affiliation: Department of Urology, Singapore General Hospital, Singapore.

ABSTRACT

Background: It is widely recognised that sorafenib inhibits a range of molecular targets in renal cell carcinoma (RCC). In this study, we aim to use patient-derived RCC xenografts to delineate the angiogenic and non-angiogenic molecular targets of sorafenib therapy for advanced RCC (aRCC).

Methods: We successfully generated three patient RCC-derived xenografts in severe combined immunodeficient mice, consisting of three different RCC histological subtypes: conventional clear cell, poorly differentiated clear cell RCC with sarcomatoid changes, and papillary RCC. This study also used clear cell RCC cells (786-0/EV) harbouring mutant VHL to investigate the clonogenic survival of cells transfected with survivin sense and antisense oligonucleotides.

Results: All three xenografts retain their original histological characteristics. We reported that sorafenib inhibited all three RCC xenograft lines regardless of histological subtypes in a dose-dependant manner. Sorafenib-induced growth suppression was associated with not only inhibition of angiogenic targets p-PDGFR-β, p-VEGFR-2, and their downstream signalling pathways p-Akt and p-ERK, cell cycle, and anti-apoptotic proteins that include cyclin D1, cyclin B1, and survivin but also upregulation of proapoptotic Bim. Survivin knockdown by survivin-specific antisense-oligonucleotides inhibited colony formation and induced cell death in clear cell RCC cells.

Conclusion: This study has shed light on the molecular mechanisms of sorafenib in RCC. Inhibition of non-angiogenic molecules by sorafenib could contribute in part to its anti-tumour activities observed in vivo, in addition to its anti-angiogenic effects.

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Related in: MedlinePlus

Effects of sorafenib on phospho-ERK1/2, VEGF expression, angiogenesis, cell proliferation and apoptosis of RCC-07-0408 xenograft. Mice bearing RCC-07-0408 tumours were randomised (10 mice per group) and treated with vehicle or 40 mg kg−1 per day sorafenib for 21 days. Representative pictures of blood vessels stained with anti-CD31, proliferative cells stained with anti-Ki-67, apoptotic cells stained with anti-cleaved-PARP, VEGF expression stained with anti-VEGF, and p-ERK1/2 stained with anti-phospho-ERK antibodies in vehicle- and drug-treated tumours are shown ( × 200). Experiments were repeated twice with similar results.
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fig3: Effects of sorafenib on phospho-ERK1/2, VEGF expression, angiogenesis, cell proliferation and apoptosis of RCC-07-0408 xenograft. Mice bearing RCC-07-0408 tumours were randomised (10 mice per group) and treated with vehicle or 40 mg kg−1 per day sorafenib for 21 days. Representative pictures of blood vessels stained with anti-CD31, proliferative cells stained with anti-Ki-67, apoptotic cells stained with anti-cleaved-PARP, VEGF expression stained with anti-VEGF, and p-ERK1/2 stained with anti-phospho-ERK antibodies in vehicle- and drug-treated tumours are shown ( × 200). Experiments were repeated twice with similar results.

Mentions: We next examined the anti-angiogenic, anti-proliferative, and apoptotic effects of sorafenib in treated tumour xenografts. Representative CD31, Ki-67, cleaved PARP, VEGF, and p-ERK1/2 immunohistochemical stainings for vehicle- and sorafenib-treated RCC-07-0408 and RCC-02-0908 tumour xenografts were shown in Figure 3. Sorafenib-treated xenografts demonstrated significant decrease in mean percentage of CD31-positive endothelial cells in both xenografts compared with vehicle-treated xenografts (P<0.05). In relation to this, there was no difference in the expression of VEGF between the sorafenib- and vehicle-treated xenografts, however, the former demonstrated significant decrease in p-ERK staining compared with the latter (P<0.05). This finding is consistent with inhibition by sorafenib of the angiogenic pathway mediated by VEGFR and PDGFR in endothelial cells. In addition, there was a significant increase in apoptosis (percentage of cleaved PARP-positive cells) and a significant decrease in proliferation (percentage of Ki-67-positive cells) (P<0.05) observed in both sorafenib-treated xenografts compared with the vehicle-treated control. Similar results were obtained when RCC-25-0908 tumours were analysed (Supplementary Figure 2S).


Inhibition of angiogenic and non-angiogenic targets by sorafenib in renal cell carcinoma (RCC) in a RCC xenograft model.

Yuen JS, Sim MY, Siml HG, Chong TW, Lau WK, Cheng CW, Huynh H - Br. J. Cancer (2011)

Effects of sorafenib on phospho-ERK1/2, VEGF expression, angiogenesis, cell proliferation and apoptosis of RCC-07-0408 xenograft. Mice bearing RCC-07-0408 tumours were randomised (10 mice per group) and treated with vehicle or 40 mg kg−1 per day sorafenib for 21 days. Representative pictures of blood vessels stained with anti-CD31, proliferative cells stained with anti-Ki-67, apoptotic cells stained with anti-cleaved-PARP, VEGF expression stained with anti-VEGF, and p-ERK1/2 stained with anti-phospho-ERK antibodies in vehicle- and drug-treated tumours are shown ( × 200). Experiments were repeated twice with similar results.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3065286&req=5

fig3: Effects of sorafenib on phospho-ERK1/2, VEGF expression, angiogenesis, cell proliferation and apoptosis of RCC-07-0408 xenograft. Mice bearing RCC-07-0408 tumours were randomised (10 mice per group) and treated with vehicle or 40 mg kg−1 per day sorafenib for 21 days. Representative pictures of blood vessels stained with anti-CD31, proliferative cells stained with anti-Ki-67, apoptotic cells stained with anti-cleaved-PARP, VEGF expression stained with anti-VEGF, and p-ERK1/2 stained with anti-phospho-ERK antibodies in vehicle- and drug-treated tumours are shown ( × 200). Experiments were repeated twice with similar results.
Mentions: We next examined the anti-angiogenic, anti-proliferative, and apoptotic effects of sorafenib in treated tumour xenografts. Representative CD31, Ki-67, cleaved PARP, VEGF, and p-ERK1/2 immunohistochemical stainings for vehicle- and sorafenib-treated RCC-07-0408 and RCC-02-0908 tumour xenografts were shown in Figure 3. Sorafenib-treated xenografts demonstrated significant decrease in mean percentage of CD31-positive endothelial cells in both xenografts compared with vehicle-treated xenografts (P<0.05). In relation to this, there was no difference in the expression of VEGF between the sorafenib- and vehicle-treated xenografts, however, the former demonstrated significant decrease in p-ERK staining compared with the latter (P<0.05). This finding is consistent with inhibition by sorafenib of the angiogenic pathway mediated by VEGFR and PDGFR in endothelial cells. In addition, there was a significant increase in apoptosis (percentage of cleaved PARP-positive cells) and a significant decrease in proliferation (percentage of Ki-67-positive cells) (P<0.05) observed in both sorafenib-treated xenografts compared with the vehicle-treated control. Similar results were obtained when RCC-25-0908 tumours were analysed (Supplementary Figure 2S).

Bottom Line: Sorafenib-induced growth suppression was associated with not only inhibition of angiogenic targets p-PDGFR-β, p-VEGFR-2, and their downstream signalling pathways p-Akt and p-ERK, cell cycle, and anti-apoptotic proteins that include cyclin D1, cyclin B1, and survivin but also upregulation of proapoptotic Bim.This study has shed light on the molecular mechanisms of sorafenib in RCC.Inhibition of non-angiogenic molecules by sorafenib could contribute in part to its anti-tumour activities observed in vivo, in addition to its anti-angiogenic effects.

View Article: PubMed Central - PubMed

Affiliation: Department of Urology, Singapore General Hospital, Singapore.

ABSTRACT

Background: It is widely recognised that sorafenib inhibits a range of molecular targets in renal cell carcinoma (RCC). In this study, we aim to use patient-derived RCC xenografts to delineate the angiogenic and non-angiogenic molecular targets of sorafenib therapy for advanced RCC (aRCC).

Methods: We successfully generated three patient RCC-derived xenografts in severe combined immunodeficient mice, consisting of three different RCC histological subtypes: conventional clear cell, poorly differentiated clear cell RCC with sarcomatoid changes, and papillary RCC. This study also used clear cell RCC cells (786-0/EV) harbouring mutant VHL to investigate the clonogenic survival of cells transfected with survivin sense and antisense oligonucleotides.

Results: All three xenografts retain their original histological characteristics. We reported that sorafenib inhibited all three RCC xenograft lines regardless of histological subtypes in a dose-dependant manner. Sorafenib-induced growth suppression was associated with not only inhibition of angiogenic targets p-PDGFR-β, p-VEGFR-2, and their downstream signalling pathways p-Akt and p-ERK, cell cycle, and anti-apoptotic proteins that include cyclin D1, cyclin B1, and survivin but also upregulation of proapoptotic Bim. Survivin knockdown by survivin-specific antisense-oligonucleotides inhibited colony formation and induced cell death in clear cell RCC cells.

Conclusion: This study has shed light on the molecular mechanisms of sorafenib in RCC. Inhibition of non-angiogenic molecules by sorafenib could contribute in part to its anti-tumour activities observed in vivo, in addition to its anti-angiogenic effects.

Show MeSH
Related in: MedlinePlus