Int J Thyroidol.  2021 Nov;14(2):127-134. 10.11106/ijt.2021.14.2.127.

Treatment Effect of Combining Lenvatinib and Vemurafenib for BRAF Mutated Anaplastic Thyroid Cancer

• Hong CM
• Oh JM
• Gangadaran P
• Rajendran RL
• Ahn BC

• KMID: 2522917
• DOI: http://doi.org/10.11106/ijt.2021.14.2.127

Abstract

Background and Objectives
Even though most of the thyroid cancer shows good prognosis, de-differentiated thyroid cancer is still refractory to conventional treatments. Recently, kinase inhibitors including multi-kinase and BRAF inhibitors are widely used for treatment of de-differentiated thyroid cancers, but resistant to single kinase inhibitor treatment eventually encountered. Therefore, combination therapy may have better therapeutic effect than single therapy for thyroid cancer. In this study, we evaluated therapeutic effect of multi-kinase and BRAF inhibitor combination to anaplastic thyroid cancer cell lines with and without BRAF mutation.
Materials and Methods
We used anaplastic thyroid cancer cell lines with BRAF ^V600E mutation (8505C) and with NRAS mutation (HTh7). Both cell lines were treated with various concentration of multi-kinase inhibitor (lenvatinib) and BRAF inhibitor (vemurafenib). And combination of various concentration of both kinase inhibitors were used to treat both cell lines. Cytotoxic effect was assessed with cell counting kit-8 and therapeutic effect of single kinase inhibitor therapy and the combination therapy was compared.
Results
Anti-proliferative effect of vemurafenib on 8505C BRAF ^V600E -mutated cells was demonstrated from 0.25 μM concentration. However, HTh7 cells with NRAS mutation represented drug resistance up to 4 μM of vemurafenib. In case of lenvatinib treatment as a multi-kinase inhibitor, 8505C and HTh7 cells showed decreased cell viability dose-dependent manner. Combination treatment with vemurafenib and lenvatinib showed synergistic cytotoxic effect in BRAF mutated 8505C cell line, even at lower concentrations.
Conclusion
Combination treatment with multi-kinase inhibitor and BRAF inhibitor showed promising therapeutic results in BRAF mutated anaplastic thyroid cancer cell line.

Keyword

Anaplastic thyroid cancer; BRAF; Lenvatinib; Vemurafenib; Combination treatment

Figure

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  Fig. 1 Anti-proliferative effect of lenvatinib and vemurafenib in HTh7 and 8505C anaplastic thyroid cancer cells. (A) Single treatment of vemurafenib in BRAFV600E mutated 8505C cells showed anti-proliferative effect from 0.25 μM concentration. However, HTh7 cells with NRAS mutation represented drug resistance up to 4 μM and abruptly cell viability was decreased from 8 μM. (B) Single treatment of lenvatinib in 8505C and HTh7 cells showed dose dependent anti-proliferative effect. Values presented here are mean value±standard deviation.

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  Fig. 2 Combination treatment of lenvatinib and vemurafenib. (A) Combination treatment of lenvatinib and vemurafenib in HTh7 cells showed only additive cytotoxic effect. (B) In 8505C cells, synergistic cytotoxic effect was observed by the combination treatment. Values presented here are mean value±standard deviation.

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  Fig. 3 DNA damage analysis using γH2A.X stain. After 72 hours of treatment, γH2A.X stain was significantly increased in the combination treatment, compared with the control and the single treatments.

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  Fig. 4 Western blot analysis of apoptosis pathway. Western blot analysis showed significantly increased cleaved PARP by the combination treatment compared with the control and the single treatments.

Reference

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