FGFR1 and PD-1 - mediated signals are involved in growth and survival of lung cancer and should be considered as targets for therapeutic approaches. Here, we report the potential efficacy of an anti-FGFR1 humanized antibody OM-RCA-01, either alone or in combination with an anti-PD-1 antibody. To assess the activity of OM-RCA-01 on FGF-mediated signaling, A549 human adenocarcinoma cells expressing FGFR1 were incubated and dosed with OM-RCA-01. Control cells were left untreated. Six hours after dosing, FGF2 was added at a concentration of 100 ng/ml. Cell growth was determined using Promega’s Cell Titer-Glo assay. Thirty-five NCr nu/nu female mice (10-12 weeks of age) were used for A549 xenotransplantation. Twenty animals with measurable tumors were pair matched and selected on day 7 after tumor inoculation. Intravenous OM-RCA-01 (30 mg/kg, N=10) or saline (Vehicle, N=10) were administered every 3 days starting on day 7 after tumor inoculation. To evaluate the efficacy of co-inhibition, patient explants (adenocarcinoma with PD-L1 expression >50%, DAKO 22C3) were obtained from surgical lung specimens, and PDX models were generated by implantation of PDX into humanized NSG mice (Jackson Laboratory, Sacramento, CA, USA) as previously described (Wang, FASEB J., 2018). Treatment with intravenous OM-RCA-01 (30 mg/kg) with pembrolizumab (10 mg/kg) or pembrolizumab alone (10 mg/kg) or saline (vehicle) every 3 days was started when the tumors reached 70 mm3 in volume. Measurements of tumor volume were performed by digital calipers every 3 days during 31 days in xenograft and PDX studies. In vitro study showed that FGF2 significantly increased proliferation of the human lung cancer cells (P<0.001). OM-RCA-01 exerted dose-dependent inhibitory effects on FGF-triggered lung cancer cell proliferation in comparison with control (P=0.01). IC50 was 9.69 mcg/mL. In the xenograft model, treatment resulted in median tumor volume of 1,048.5 mm3 and 2,174 mm3 at day 31 in the OM-RCA-01 and vehicle groups, respectively (P<0.0001). All tumors reached the volume endpoint of 2,000 mm3 to day 31 in vehicle group in comparison with study group where maximum size of tumor was 1,309 mm3. In the PDX study, the differences between groups were also statistically significant (all P<0.001). Median tumor volume was 97.1 mm3 in the combination group (N=7), 417.1 mm3 in the pembrolizumab only group (N=7), and 863.5 mm3 in the vehicle group (N=7). No group mean body weight losses or clinical manifestations of toxicity were observed in both groups. In conclusion, OM-RCA-01 antibody has potent antitumor activity against lung cancer in vitro and in vivo. Our findings also support that the co-inhibition of FGFR1 and immune checkpoints can enhance antitumor activity compared to anti-PD-1 antibody monotherapy. Citation Format: Ilya Tsimafeyeu, Jonathan Smith, Wei Yin, Alex Fanelli, Dmitry Khochenkov, Evgenia Stepanova. Combined FGFR1 and PD-1 inhibition could enhance antitumor activity in lung cancer models [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 4165.

In tumor angiogenesis, two main mechanisms are officially recognized. They are: the formation of vascular structures by associations of endothelial cell precursors (PEK) or angioblasts from circulating peripheral blood (1) and vascular co-optation (2) using preexistent vessels. Determining the dominant phenotype of endothelial cells (EC) involved in the neoangiogenesis of uveal melanoma (UM) is very important due to the different sensitivity of different EC to antiangiogenic therapy and their biological differences. Circulating hemangioblasts of bone marrow origin (synonym: endothelial cell precursors, PEC) involved in the formation of vessels in the tumor can also originate from hematopoietic stem cells and, in conditions of an excess of proangiogenic growth factors, undergo “reverse differentiation” or “dedifferentiation”. This situation is extremely dangerous due to changes in the biological properties of EC: the rate of proliferation increases and high resistance to any type of interventional antiangiogenic therapy, including targeted antiangiogenic drugs, develops. In combination with high resistance to radiation therapy and chemotherapy of tumor cells, such a tumor becomes uncontrollable and quickly leads to death... The purpose of this study: to identify the phenotype of endothelial cells involved in UM angiogenesis by immunohistochemical analysis. The authors conducted immunohistochemical studies on paraffin blocks (n = 98) of enucleated eyes with UM. One of the recommended PEC markers was used: a monoclonal antibody to the VEGFR2-KDR/Flk-1 receptor. It was found that the expression of the VEGFR- 2/Flk-KDR-1 receptor took place only in 22.1 % of the UM. The receptor was expressed exclusively by endothelial cells (EC) of tumor vessels and was not expressed by UM cells. There was no basic expression of the receptor (at stage T1). Peak expression was observed at stage T2. At the next stages of the oncological process, there was a decline in expression. The mean IHC gradation of expression intensity also decreased from stage T2 to stage T4. Flk-positive and negative endothelial cells differed in size, shape, nature of communication with the vascular wall and the presence or absence of a nucleus in the cell. The authors concluded that both mechanisms of angiogenesis take place in UM. A small percentage (22.1 %) Flk+endothelial cells indicate that the dominant mechanism of angiogenesis in UM is co-optation of vessels based on preexistent, and the dominant population of EC is Flk-negative EC.

Thanks to new fundamental data, our understanding of the angiogenesis’ role and its molecular participants has changed. Molecular genetic mechanisms of activation of signaling pathways of proliferation, malignancy, suppression of the tumor cells’ apoptosis, carried out through the interaction of the VEGF molecule with its receptors, have been deciphered. Molecular genetic mechanisms of activation of signaling pathways of proliferation, malignancy, suppression of apoptosis of tumor cells, carried out through the interaction of the VEGF molecule with its receptors, have been deciphered. The doctrine of angiogenesis has changed. Angiogenesis began to be considered in the aspect of the increasing anaplasia mechanism, accelerating proliferation, the formation of a clone of stem tumor cells, highly resistant to chemotherapy and radiation therapy with a high potential for metastasis. It is time to reconsider the role of individual biological markers of angiogenesis in their suitability in predicting disease outcome and evaluating them as a potential target for targeted therapy. In this aspect, uveal melanoma (UM) as a model of an extremely aggressive malignant tumor using its angiogenic phenotype to accelerate hematogenous metastasis is of particular interest. One of the characteristics of the angiogenic phenotype is VEGF-R1/Flt-1. The purpose: to study the expression of the VEGF-R1/Flt-1 receptor as a characteristic of the angiogenic phenotype of UM in correlation with its clinical and morphological indicators and the outcome of the disease. The authors conducted a retrospective study on 98 archival paraffin blocks of the eyes of patients with UM. The following general patterns of Flt expression in UM cells are revealed: Basic expression takes place in the smallest tumor proliferates UM (T1). Moreover, overexpression (IGH-gradation III) of Flt in the nucleus (39.7 %) and cytoplasm (36.3 %) occurred in every third patient with UM in stage T1. The peak of reception for the maximum average percentage of positive cells and the average cellular IGH gradation of expression was recorded at the T2 stage, after which there was a slow decline to the T4 stage. The number of overexpressive Flt in the nucleus and cytoplasm of UM cells increased x 2 times (75.3 %) to the T2 stage. The peak of Flt nuclear expression in terms of the number of immunopositive cells was also recorded at the T2 stage. The authors concluded that VEGF-R1/Flt-1 expression is a very important characteristic of the UM angiogenic phenotype. In the vast majority of UM, there is an expression of the VEGF-R1/Flt-1 receptor in the nucleus and cytoplasm of tumor cells. The revealed correlations of VEGF-R1/Flt-1 expression with the volume and histological type of tumor, disease stage and metastasis allow them to consider Flt-1 an important indicator associated with the pathogenesis and prognosis of uveal melanoma and a potential target for targeted therapy. A prognostic adverse factor in the aspect of the prognosis of the risk of metastases should be considered the index of the ratio Flt C/Flt N ≥ 3.

Благодаря появившимся новым фундаментальным данным наши представления о роли ангиогенеза и его молекулярных участниках изменились. Расшифрованы молекулярно-генетические механизмы активации сигнальных путей пролиферации, малигнизации, подавления апоптоза опухолевых клеток, осуществляемые посредством взаимодействия молекулы VEGF с ее рецепторами. Сменилась доктрина ангиогенеза. Ангиогенез стали рассматривать в аспекте механизма повышения анаплазии, ускорения пролиферации, формирования клона стволовых опухолевых клеток, высокорезистентных к химиотерапии и лучевой терапии, с высоким потенциалом к метастазированию. Настало время для пересмотра роли отдельных биологических маркеров ангиогенеза в контекcте их пригодности в прогнозировании исхода заболевания и оценки в качестве потенциальной мишени для таргетной терапии. В этом аспекте увеальная меланома (УМ) как модель чрезвычайно агрессивной злокачественной опухоли, использующей свой ангиогенный фенотип для ускорения гематогенного метастазирования, привлекает особый интерес. В качестве одной из характеристик ангиогенного фенотипа служит рецептор VEGF-R1/Flt-1. Цель работы: изучить экспрессию рецептора VEGF-R1/Flt-1 как характеристику ангиогенного фенотипа УМ в корреляции с ее клинико-морфологическими показателями и исходом заболевания. Материал и методы. Авторы провели ретроспективное исследование на 98 архивных парафиновых блоках глаз пациентов с УМ. Выявлены следующие общие закономерности экспрессии Flt в клетках УМ: Базовая экспрессия имеет место при самых маленьких опухолевых пролифератах УМ (Т1). Причем гиперэкспрессия (ИГХ-градация III) Flt в ядре (39,7 %) и цитоплазме (36,3 %) имела место у каждого третьего пациента с УМ в стадии Т1. Пик рецепции по максимальному среднему, процент позитивных клеток (Ммах ср. ± м) и усредненной клеточной ИГХ-градации экспрессии зафиксирован на стадии T2, после этого происходил медленный спад до стадии T4. Число гиперэкспрессирующих Flt в ядре и цитоплазме клеток УМ возрастало в 2 раза (75,3 %) к стадии Т2. Пик ядерной экспрессии Flt по количеству иммунопозитивных клеток зафиксирован также на стадии Т2. Авторы пришли к выводу, что экспрессия VEGF-R1/Flt-1 является очень важной характеристикой ангиогенного фенотипа УМ. В подавляющем большинстве УМ имеет место экспрессия рецептора VEGF-R1/Flt-1 в ядре и цитоплазме опухолевых клеток. Выявленные корреляционные связи показателей экспрессии VEGF-R1/Flt-1 с объемом и гистологическим типом опухоли, стадией заболевания и метастазированием позволяют им считать Flt-1 важным показателем, ассоциированным с патогенезом и прогнозом увеальной меланомы и потенциальной мишенью для таргетной терапии. Прогностически неблагоприятным фактором в аспекте прогноза риска развития метастазов следует считать индекс соотношения Flt C/Flt N ≥ 3. Ключевые слова: увеальная меланома, ангиогенез, рецептор VEGF-R1/Flt-1, иммуногистохимический анализ, прогноз Офтальмология. 2021;18(4):866–875 поступила 10.08.2021 was received 10.08.2021

. The problem of creating antitumor drugs with new mechanisms of action that predominantly induce apoptosis is still topical. The extract of Gratiola officinalis is a potential antitumor agent containing mainly flavonoids. The aim of this research is to study the effects of Gratiola officinalis extract on activation of apoptosis and autophagy in breast adenocarcinoma SK-BR-3 and kidney carcinoma A-498 lines. Apoptotic activity of the extract was studied by flow cytofluorometry using Hoechst stain and double staining with annexin V plus propidium iodide. There was 96.3% of cells in SK-BR-3 culture in late apoptosis phase detected by flow cytofluorometry method at the extract concentration of 0.88 mg/ml, 86.3% of cells were in apoptosis by Hoechst stain. The concentration of 0.82 mg/ml caused apoptosis in half of the cells. The extract has cytoprotective activity at low concentration (0.0352 mg/ml). The cytoprotection mechanism is realized through the activation of autophagy. The maximum number of autophagosomes in kidney carcinoma cells is observed at the extract concentration of 0.056 mg/ml. Thus, Gratiola officinalis extract is able to block cytoprotective autophagy with increasing the extract concentration and to activate apoptosis in 85% of tumor cells. Detailed research should be continued to understand the mechanisms of antitumor activity of Gratiola officinalis extract.

A prerequisite for the growth, progression and metastasis of malignant tumors of any localization is the development of its own vascular network. Newly formed vessels not only nourish the primary tumor, but also create conditions for the spread of tumor cells through the circulatory system and the formation of distant metastases. Angiogenesis is able to launch a small population of tumors from 100–300 cells that have accumulated genetic aberrations and have begun to express proangigenic molecules. The phenomenon is known as “transformation of tumor cells into angiogenic phenotype”. A tumor with angiogenic phenotype carries a high potential for proliferation and malignization. This pattern has been found in many types of cancer, but studied less in uveal melanoma. Meanwhile, in this aspect, uveal melanoma, metastasizing exclusively in a hematogenic way, with its selective, organotropic nature of metastasis, becomes an attractive model for the study of the molecular “scenario” of tumor angiogenesis studies allow us to say that, UM is subject to the general patterns of the development of malignant tumors. As with many types of tumors, VEGF is an obligate condition for the development and progression of UM. The VEGF molecule’s producers in UM are two cell populations: endothelial vascular cells and tumor cells. VEGF’s expression in UM is cyclical. The cycle is re-initiated, apparently, by increasing cell density in tumor proliferate and the development of hypoxia zones. We found no correlation between the intensity of pigmentation, necrosis, hemorrhage, germination in the corner of the front chamber, ophthalmohypertension on the one hand, and expression of VEGF in UM cells on the other. At the same time, a direct link between the expression of VEGF in tumor cells and EC vessels on the one hand and the thickness, base diameter, as well as the localization of UM, on the other hand, has been revealed. Additionally, VEGF expression in tumor cells was closely correlated with the histological structure of UM, and VEGF expression in EC correlated with the stage of the disease. Thus, the authors showed that UM, like other malignant solid tumors, is prone to transformation into angiogenic phenotype and expression of VEGF.

Novel 4-amino-5-cyano-2-sulfonylpyrimidines were prepared based on three-component cyclization between isothiouronium salts, benzaldehydes and malononitrile, followed by oxidation of the sulfide moiety with Oxone. The cytotoxic activity of the synthesized compounds, as well as the induction of apoptosis, inhibition of the cell cycle and proliferation tests were performed on selected cancer cell lines A431, A549, A375, HCT 116, MCF7, LNCap and SH-SY5Y.

Renal cell carcinoma (RCC) ranks first in mortality among urogenital tumors and is the most common disease after prostate and bladder cancer. Early detection of RCC allows immediately undertaking appropriate treatment, which significantly increases the survival of patients. In the case of the asymptomatic RCC, timely diagnosis in the early stages is usually difficult. To date, the problem of searching for molecular markers of clear cell RCC, which allows to determine the stage, metastatic potential and prognosis of disease, or select a treatment regimen remains topical. Of particular interest are early-stage biomarkers of RCC and its metastatic potential, as well as markers that can be obtained by non-invasive or minimally invasive methods. This review presents modern methods for diagnosing RCC using biomarkers.

Melanoma is one of the most aggressive and treatment-resistant tumors that responsible for majority of skin-cancer related deaths. Here we propose a combination of MEK inhibitor binimetinib with metformin as a promising therapy against human melanoma cells in vitro, including BRAF -mutated A375, Mel Z, and Mel IL cells, and NRAS-mutated Mel MTP and Mel Me cells. Additionally, we obtained two close to clinical practice models of melanoma progression. The first one was vemurafenib-resistant Mel IL/R melanoma cells with acquired resistance to BRAF inhibition-targeted therapy, and the second one was tumor spheroids, which are 3D in vitro model of small-size solid tumors in vivo. The cytotoxicity of binimetinib and metformin was synergistic in both 2D and 3D melanoma culture and mediated through apoptotic pathway. The combination reduced the number of melanoma-formed colonies, inhibited cell invasion and migration, and led to G0/G1 cell cycle arrest through cyclin D/CDK4/CDK6 pathway. The mechanism of metformin and binimetinib synergy in melanoma cells was associated with increased activation of p-AMPKα and decreased p-ERK, but not with alterations in p-mTOR. In summary, the combination of metformin and binimetinib resulted in stronger anti-proliferative effects on melanoma cells compared to binimetinib alone, and therefore could be promising for clinical applications.

The incidence of malignant melanoma is increasing. The discovery of agents specifically targeting the mutated cascades has provided a good response for patients with oncogenic B-Raf proto-оncogene, serine/threonine kinase (BRAF). However, numerous studies continue to focus on novel methods of treatment to overcome acquired resistance to novel drugs. Recently, it has been revealed that inhibition of endoplasmic reticulum (ER) stress chaperon 78 kDa glucose-regulated protein 78 (GRP78) leads to down-regulation of autophagy and increased sensitivity to temozolomide (TMZ) treatment. Melanoma cells have a different sensitivity to TMZ treatment, which corresponds to the basal autophagy level. In the present study, we demonstrated that downregulation of GRP78 mitigated chemoresistance to TMZ in three melanoma cell lines. We found that downregulation of GRP78 led to inhibition of autophagy, cell cycle arrest in the G0/G1 phase, and activation of caspase-7-induced apoptosis, and this was affected by the initial autophagy level. Moreover, inhibition of GRP78 mitigated the combined TMZ and chloroquine effect. Our data revealed that autophagy inhibition through downregulation of ER stress response could overcome resistance to TMZ treatment in melanoma cells with a high basal level of autophagy treatment, which makes this combination a potential potent antitumor treatment for metastatic melanoma.

Objective ― The discovery of the apoptosis-inducing effects of flavonoid vagonin allowed to make an assumption of existence of similar effect in others flavonoids. This study is devoted to the effects of Gratīola officinālis extract on cell culture of the human kidney cancer. Methods ― Cell cultures of human kidney carcinoma – Caki-1 and SN12c were used in the study. The cells were stained with Hoechat 33258 dye. The number of living cells, cells in a state of apoptosis and mitosis were counted. The Cramer-Welch criterion (T) was used to compare the obtained data. Results ― The activation of apoptosis was noted at all concentrations of the Gratiola officinalis L. extract during the first day of exposure. The apoptotic activity increased with increasing of extract concentration. After 48 hours, this activity was maintained only at a Gratiola officinalis L. extract concentration of 0.9 mg/ml. After 24 hours, the apoptotic activity of the extract was more expressed in the culture of CaKi-1. However, after 48 hours the extract induces more pronounced apoptosis in the culture of Sn12c cells. The cytotoxic activity of the extract was not differ after 24 hours in both cultures, after 48 hours it was more pronounced in the culture of CaKi-1. Conclusion ― We revealed a pronounced antitumor and apoptotic activity of the Gratiola officinalis L. extract against the cultures of the kidney cancer Caki-1 and Sn12c. Apoptosis of tumor cells can be manifested in the form of pycnosis of the nucleus, the formation of apoptotic bodies and the emergence of cellular debris resulting from complete degradation of tumor cells. The apoptotic activity of the Gratiola officinalis extract depends on their concentration.

e12562 Background: A few prognostic factors are used to make decision on breast cancer (BC) treatment. Researches to identify novel prognostic and predictive factors are still actual. Two DNA repair proteins were studied as prognostic markers for patients with early HER2-positive BC, who received or not adjuvant trastuzumab (T). Methods: 103 patients with HER2+ BC, I-IIIa stage, treated with surgery in 3 Russian Cancer Centers . from 2002 to 2007 were included in analysis. Adjuvant treatment with trastuzumab was performed for 58 patients (56,3%) only. In all cases expression of ERCC1 and XRCC1 were immunohistochemistry studied. Patient characteristics at the time of BC diagnosis, expression of biomarkers, treatment options and their associations with overall survival (OS) and progression free survival (PFS) were evaluated by Kaplan-Meier curves, log-rank tests, and Cox proportional hazard models. Results: 8-years PFS was 74,4%: 80,3 % for patients received adjuvant trastuzumab vs 60,9 % for patient without anti-HER treatment (p < 0,0001). 8-years OS was 81,9%: 88,4% vs 59,5% (p < 0,0001), respectively. Overexpression of XRCC1 was detected in 61,2% of patients and significantly correlated with poor PFS [HR = 7,181, 95% CI 2,149-24,00; p = 0,001]. The most significant differences were registered in target-therapy group: 8-years PFS for XRCC1-positive patients (n = 32) was 55,4% vs 96,2% for XRCC1-negative patients (n = 26) received trastuzumab (p < 0,0001). Overexpression of ERCC1 was detected in 65% of patients and significantly correlated with poor PFS [HR = 2,179, 95% CI 1,123-6,581; p = 0,027] and OS [HR = 3,071, 95% CI 1,017-9,269; p = 0,047] either. Adjuvant trastuzumab therapy significantly proof PFS and OS only in ERCC1-negative patients (p = 0,008 and p = 0,009, respectively). ERCC1-positive patients had poor prognosis independent of adjuvant T usage: 8-years PFS was 56,3% in trastuzumab group (n = 31) vs 73,1% in group without T (n = 36), p = 1,0; 8-y OS was 74,6% vs 76,8%, respectively, p = 0,45. Conclusions: Overexpression of XRCC1 and ERCC1 predict poor prognosis in early HER2-positive BC. We consider ERCC1 should be additional studied as a marker of trastuzumab resistance

The authors regret that one of the co-authors names was misspelled in the original article. Dmitry Kochenkov should be spelt Dmitry Khochenkov. The authors would like to apologise for any inconvenience caused. Targeting FGFR2 with alofanib (RPT835) shows potent activity in tumour modelsEuropean Journal of CancerVol. 61PreviewAlofanib (RPT835) is a novel selective allosteric inhibitor of fibroblast growth factor receptor 2 (FGFR2). We showed previously that alofanib could bind to the extracellular domain of FGFR2 and has an inhibitory effect on FGF2-induced phoshphorylation of FRS2α. In the present study, we further showed that alofanib inhibited phosphorylation of FRS2α with the half maximal inhibitory concentration (IC50) values of 7 and 9 nmol/l in cancer cells expressing different FGFR2 isoforms. In a panel of four cell lines representing several tumour types (triple-negative breast cancer, melanoma, and ovarian cancer), alofanib inhibited FGF-mediated proliferation with 50% growth inhibition (GI50) values of 16–370 nmol/l. Full-Text PDF

// Marina Emelyanova 1 , Lilit Ghukasyan 1 , Ivan Abramov 1, 2 , Oxana Ryabaya 2 , Evgenia Stepanova 2 , Anna Kudryavtseva 1, 3 , Asiya Sadritdinova 1, 3 , Cholpon Dzhumakova 2 , Tatiana Belysheva 2 , Sergey Surzhikov 1 , Lyudmila Lyubchenko 2 , Alexander Zasedatelev 1, 2 and Tatiana Nasedkina 1, 2 1 Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Moscow, Russian Federation 2 Blokhin Cancer Research Center, Ministry of Health of the Russian Federation, Moscow, Russian Federation 3 P. Hertsen Moscow Oncology Research Institute, Moscow, Russian Federation Correspondence to: Marina Emelyanova, email: emel_marina@mail.ru Keywords: biochip, somatic mutations, melanoma, diagnostic tool, targeted therapy Received: December 30, 2016 Accepted: March 30, 2017 Published: April 10, 2017 ABSTRACT Target inhibitors are used for melanoma treatment, and their effectiveness depends on the tumor genotype. We developed a diagnostic biochip for the detection of 39 clinically relevant somatic mutations in the BRAF , NRAS , KIT , GNAQ , GNA11 , MAP2K1 and MAP2K2 genes. We used multiplex locked nucleic acid (LNA) PCR clamp for the preferable amplification of mutated over wild type DNA. The amplified fragments were labeled via the incorporation of fluorescently labeled dUTP during PCR and were hybridized with specific oligonucleotides immobilized on a biochip. This approach could detect 0.5% of mutated DNA in the sample analyzed. The method was validated on 253 clinical samples and six melanoma cell lines. Among 253 melanomas, 129 (51.0%) BRAF , 45 (17.8%) NRAS , 6 (2.4%) KIT , 4 (1.6%) GNAQ , 2 (0.8%) GNA11 , 2 (0.8%) MAP2K1 and no MAP2K2 gene mutations were detected by the biochip assay. The results were compared with Sanger sequencing, next generation sequencing and ARMS/Scorpion real-time PCR. The specimens with discordant results were subjected to LNA PCR clamp followed by sequencing. The results of this analysis were predominantly identical to the results obtained by the biochip assay. Infrequently, we identified rare somatic mutations. In the present study we demonstrate that the biochip-based assay can effectively detect somatic mutations in approximately 70% of melanoma patients, who may require specific targeted therapy.