Treatment of Vemurafenib-Resistant SKMEL-28 Melanoma Cells with Paclitaxel

Nguyen, Dinh Thang,Phan, Tuan Nghia,Kumasaka, Mayuko Y.,Yajima, Ichiro,Kato, Masashi Asian Pacific Journal of Cancer Prevention 2015 Asian Pacific journal of cancer prevention Vol.16 No.2

Vemurafenib has recently been used as drug for treatment of melanomas with $BRAF^{V600E}$ mutation. Unfortunately, treatment with only vemurafenib has not been sufficiently effective, with recurrence after a short period. In this study, three vemurafenib-resistant $BRAF^{V600E}$ melanoma cell lines, $A375P^R$, $A375M^R$ and SKMEL-$28^R$, were established from the original A375P, A375M and SKMEL-28 cell lines. Examination of the molecular mechanisms showed that the phosphorylation levels of MEK and ERK, which play key roles in the RAS/RAF/MEK/ERK signaling pathway, were reduced in these three cell lines, with increased phosphorylation levels of pAKTs limited to SKMEL-$28^R$ cells. Treatment of SKMEL-$28^R$ cells with 100 nM paclitaxel resulted in increased apoptosis and decreased cellular proliferation, invasion and colony formation via reduction of expression levels of EGFR and pAKTs. Moreover, vemurafenib-induced pAKTs in SKMEL-$28^R$ were decreased by treatment with an AKT inhibitor, MK-2206. Taken together, our results revealed that resistance mechanisms of $BRAF^{V600E}$-mutation melanoma cells to vemurafenib depended on the cell type. Our results suggested that paclitaxel should be considered as a drug in combination with vemurafenib to treat melanoma cells.

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

홍채문,오지민,GANGADARAN PRAKASH,Ramya Lakshmi Rajendran,안병철 대한갑상선학회 2021 International Journal of Thyroidology Vol.14 No.2

Background and Objectives: Even though most of the thyroid cancer shows good prognosis, de-differentiatedthyroid cancer is still refractory to conventional treatments. Recently, kinase inhibitors including multi-kinase andBRAF inhibitors are widely used for treatment of de-differentiated thyroid cancers, but resistant to single kinaseinhibitor treatment eventually encountered. Therefore, combination therapy may have better therapeutic effectthan single therapy for thyroid cancer. In this study, we evaluated therapeutic effect of multi-kinase and BRAFinhibitor combination to anaplastic thyroid cancer cell lines with and without BRAF mutation. Materials andMethods: We used anaplastic thyroid cancer cell lines with BRAFV600E mutation (8505C) and with NRAS mutation(HTh7). Both cell lines were treated with various concentration of multi-kinase inhibitor (lenvatinib) and BRAFinhibitor (vemurafenib). And combination of various concentration of both kinase inhibitors were used to treatboth cell lines. Cytotoxic effect was assessed with cell counting kit-8 and therapeutic effect of single kinaseinhibitor therapy and the combination therapy was compared. Results: Anti-proliferative effect of vemurafenibon 8505C BRAFV600E-mutated cells was demonstrated from 0.25 μM concentration. However, HTh7 cells withNRAS mutation represented drug resistance up to 4 μM of vemurafenib. In case of lenvatinib treatment as amulti-kinase inhibitor, 8505C and HTh7 cells showed decreased cell viability dose-dependent manner. Combinationtreatment 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 inhibitorshowed promising therapeutic results in BRAF mutated anaplastic thyroid cancer cell line.

P250 : Low-concentration vemurafenib induces the proliferation and invasion of human HaCaT keratinocytes through MAPK pathway activation

( Mi Ryung Roh ),( Jung Min Kim ),( Sang Hee Lee ),( Hong Sun Jang ),( Kyu Hyun Park ),( Kee Yang Chung ),( Sun Young Rha ) 대한피부과학회 2014 대한피부과학회 학술발표대회집 Vol.66 No.2

Background: Cutaneous squamous cell carcinomas and keratoacanthomas commonly occur in patients treated with BRAF inhibitors. Objectives: We investigated the effect of BRAF inhibitor vemurafenib on normal immortalized human HaCaT keratinocytes to explore the mechanism of hyperproliferative cutaneous neoplasia associated with the use of BRAF inhibitors. Methods: Human Melanoma cell lines SK-MEL-24(BRAF V600E mutation), G361(BRAF V600E mutation), SK-MEL-2(NRAS Q61R mutation) and HaCaT were cultured. Cell viability assay, immunoblotting, proliferation assay, Matrigel transwell assay, and gelatin zymography was performed to evaluate the effect of vemurafenib on the cell lines. Results: Vemurafenib induced an increase in viable cell number in BRAF wild-type cell lines but not in BRAF mutant cell lines. In HaCaT keratinocytes, a low concentration(2μM) of vemurafenib increased cell proliferation and activated MEK/ERK in a CRAF-dependent manner. Invasiveness of HaCaT cells in a Matrigel assay significantly increased upon cultivation of cells with 2μM vemurafenib for 24h. Gelatin zymography revealed that 2μM vemurafenib increased activated MMP-2 expression in both cytosol and conditioned media while MMP-9 expression increased in cytosol. Conclusion: These results offer additional insight into the complex mechanism of paradoxical MAPK signaling involved in hyperproliferative cutaneous neoplasia that arise after BRAF inhibition and suggest a possible role for MMPs in tumor progression and invasion.

Two cases of keratoacanthoma associated B-RAF enzyme inhibitor (Vemurafenib)

( Chan Seong Park ),( Donghwi Jang ),( Jongeun Lee ),( Hyun Jeong Byun ),( Youngkyoung Lim ),( Ji-hye Park ),( Jong Hee Lee ),( Dong-youn Lee ),( Joo-heung Lee ),( Jun-mo Yang ) 대한피부과학회 2019 대한피부과학회 학술발표대회집 Vol.71 No.1

Keratoacanthoma is a unique skin tumor which shows similar histopathologic features to SCC, but regresses voluntarily after rapid growth. Recently, there have been cases in which secondary cancer has been induced by a new anti-cancer drug. Vemurafenib is used for treatment of metastatic and unresectable melanomas with V600E mutations in the BRAF gene. BRAF is a component of the mitogen-activated protein kinase (MAPK) pathway, leading to RAS-RAF-MEK-ERK which is involved in cell proliferation, survival, and differentiation. Mutations in the BRAF gene cause a permanent activation of B-RAF and by extension, of the MAPK pathway, which results in uncontrolled growth and proliferation of cells. Vemurafenib, small-molecule inhibitor of BRAF(V600E) kinase, cause an inhibition of an over-activated MAPK signaling pathway downstream in BRAF kinase-expressing tumor cells. Proliferation of keratinocytes is common when using Vemurafenib, ranging from benign verrucous lesions to malignant. It usually occurs between the 7th and 8th weeks of treatment. In our two cases, a 58-year-old man and a 66-year-old woman, who had metastatic melanoma, were referred because of keratotic papules and nodules on face and neck. Both patients were treated with Vemurafenib for two and three months. We performed skin biopsy and the skin lesions of the both patients were diagnosed as keratoacanthoma. We report two cases of B-RAF enzyme inhibitor associated keratoacanthoma.

Vemurafenib Enhances NK cell Expansion and Tumor-killing Activity for Cancer Immunotherapy

신민화 대한의생명과학회 2023 Biomedical Science Letters Vol.29 No.4

Natural killer (NK) cells are innate immune cells and play important roles as the first immune cells to recognize and kill cancer. In patients with advanced and terminal cancer, NK cells are often inactivated, suggesting that NK cells may play important roles in cancer treatment. In particular, the proportion of NK cells among immune cells infiltrating tumor tissues is often low, which suggests that NK cells do not survive in tumor microenvironment (TME). In order to overcome these hurdles of NK cells in cancer treatment, it is critical to develop strategies that enhance the proliferation and cytolytic activity of NK cells. We applied Vemurafenib to NK cells and measured the degree of NK cell proliferation and functional activation. We obtained unexpected results of increased NK cell numbers and anti-tumor activity after Vemurafenib treatment. Although further investigation is required to uncover the detailed mechanisms, our results suggest that Vemurafenib is a promising candidate to increase the efficacy of cancer immunotherapy using NK cells.

Improvement of Neurodegenerative Disease after Use of Vemurafenib in Refractory BRAF V600E-Mutated Langerhans Cell Histiocytosis: A Case Report

Young Kwon Koh,Su Hyun Yoon,Sung Han Kang,Hyery Kim,Ho Joon Im,Pyeong Hwa Kim,Ah Young Jung,Kyung-Nam Koh 대한소아혈액종양학회 2022 Clinical Pediatric Hematology-Oncology Vol.29 No.2

Langerhans cell histiocytosis (LCH) is a rare histiocytic disorder characterized by het-erogenous lesions infiltrated with CD1a+/CD207+ cells. Although LCH has a relatively good prognosis, the prognosis for patients with LCH refractory to standard chemo-therapy is poor. Neurodegenerative LCH (ND-LCH) is a central nervous system com-plication of LCH that is characterized by progressive radiological and clinical abnormalities. Symptomatic ND-LCH is difficult to treat and therefore has a poor prognosis. A two-year-old boy presented with a scalp mass. Biopsy confirmed LCH. Whole-body imaging revealed LCH involvement in multiple bones of the skull, facial bones, and lungs. Prednisolone and vinblastine chemotherapy was initiated. One-year post-treatment, most of the lesions in the bones and lung nodules dis-appeared, and chemotherapy was discontinued. New neurodegenerative lesions ap-peared 4 months after chemotherapy was discontinued. Second-line chemotherapy using cytarabine, vincristine, and prednisolone was initiated. However, neurological manifestations of ND-LCH worsened post second-line treatment, and the treatment was switched to cytarabine and cladribine. Despite third-line chemotherapy, the le-sions progressed, and neurological deficits worsened. After identifying BRAF V600E mutation in the tumor tissue using next-generation sequencing, cytotoxic chemo-therapy was discontinued and vemurafenib treatment was initiated. One-year post-vemurafenib therapy, ND-LCH manifestations regressed, and the patient experi-enced neurological improvement.

RAF 돌연변이 대장암 세포주에서 5-fluorouracil과 표적항암제의 순서 의존적 병용 처치에 의한 상승적 세포 사멸 효과

김도형(Do Hyung Kim),장혜연(Hye Yeon Jang),MD Abdullah,이승진(Seung Jin Lee) 대한약학회 2018 약학회지 Vol.62 No.6

서 론(Introduction) 실험방법(Experimental Methods) 결과 및 고찰(Results and Discussion) 결 론(Conclusion) 감사의 말씀(Acknowledgments)

Therapeutic Inhibitors against Mutated BRAF and MEK for the Treatment of Metastatic Melanoma

류순효,윤차경,문애란,Amanda Howland,Cheryl A. Armstrong,Peter I Song 전남대학교 의과학연구소 2017 전남의대학술지 Vol.53 No.3

Melanoma is one of the most aggressive cancers in the world and is responsible for the majority of skin cancer deaths. Recent advances in the field of immunotherapy using active, adoptive, and antigen-specific therapeutic approaches, have generated the expectation that these technologies have the potential to improve the treatment of advanced malignancies, including melanoma. Treatment options for metastatic melanoma patients have been dramatically improved by the FDA approval of new therapeutic agents including vemurafenib, dabrafenib, and sorafenib. These kinase inhibitors have the potential to work in tandem with MEK, PI3K/AKT, and mTOR to inhibit the activity of melanoma inducing BRAF mutations. This review summarizes the effects of the new therapeutic agents against melanoma and the underlying biology of these BRAF inhibitors.

P419 : A case of treatment of BRAF-mutated malignant melanoma

( Nam Hee Sung ),( Tae Han Kim ),( Do Hun Kim ),( Won Suk Lim ),( Seung Ho Lee ),( Ai Young Lee ),( Hyo Seung Shin ) 대한피부과학회 2014 대한피부과학회 학술발표대회집 Vol.66 No.1

진행성 악성 흑색종(Malignant Melanoma)의 새로운 내과적 치료

이수정 ( Soo Jung Lee ),채의수 ( Yee Soo Chae ) 대한내과학회 2013 대한내과학회지 Vol.85 No.4

현재까지의 결과를 종합해 볼 때 악성 흑색종의 치료는 근치적 절제이다. 수술적 치료가 불가능한 전이성 흑색종의 경우 최근 다양한 표적치료제 및 면역요법의 임상 결과가 발표되면서 생존율의 향상을 가져왔다. 최근의 분자생물학의 발전으로 악성 흑색종을 분자생물학적·유전적 특성에 따라 세분화하여 이에 따른 적절한 표적치료제의 사용으로 이제까지 예후가 아주 불량한 것으로 여겨왔던 악성 흑색종에서도 환자의 생명연장을 기대할 수 있게 되었다. 다만 국내에서는 dacarbazine 단독 또는 병합요법이 여전히 1차 약제로 규정하고 있으며 새로이 개발된 표적치료제가 국내에 시판되어 있지 않거나 고가이어서 임상에서 사용하는 데 여전히 제한점이 많이 있다. 최근의 대규모 3상 임상시험 결과를 바탕으로 한 국내 보험 규정의 수정과 의료행정을 담당하는 정부부처와의 긴밀한 토의를 통하여 가능한 빠른 시일 안에 흑색종 환자들이 치료의 혜택을 받을 수 있도록 하는 노력이 절실히 필요하다.