Progress of BRAF Gene Alteration in Non-small Cell Lung Cancer / 中国肺癌杂志

V-Raf murine sarcoma viral oncogene homolog B (BRAF) alteration is one of the most essential driver genes of non-small cell lung cancer (NSCLC). BRAF encodes serine/threonine protein kinases, and its mutations typically lead to protein compositional activation, thereby activating the mitogen-activated protein kinase kinase (MEK) signaling pathway. A promising new approach for the treatment of mutated BRAF and/or downstream MEK may provide customized treatment opportunities for BRAF driven NSCLC patients. However, combination therapy is necessary to overcome the difficulties such as short duration of benefit, poor therapeutic effect of non-V600 BRAF mutations and susceptibility to drug resistance. This article reviewed the progress in structural characteristics, related signaling pathways, mutation types of BRAF gene, and the clinical pathological relationship between BRAF mutations and NSCLC, as well as the therapy, in order to provide more evidences for clinical doctors to make treatment decisions.
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Diagnostic value of contrast-enhanced ultrasound combined with fine-needle aspiration biopsy and BRAF gene detection for TI-RADS category 4 nodules / 中华超声影像学杂志

Objective:To explore the diagnostic value of contrast-enhanced ultrasound combined with fine-needle aspiration biopsy and BRAF gene detection for TI-RADS category 4 nodules.Methods:The clinical datas of 80 patients who underwent surgery in the First Affiliated Hospital, Zhejiang University School of Medicine and Lishui People′s Hospital and diagnosed with TI-RADS 4 thyroid nodules from January 2019 to January 2020 were retrospectively analyzed. All patients received contrast-enhanced ultrasound combined fine-needle aspiration biopsy and BRAF gene detection, the ROC curves were plotted, the area under the ROC curve(AUC) and the best diagnostic cut-off values were calculated, and the application value of ultrasound-enhanced contrast, fine-needle aspiration biopsy and BRAF gene detection were compared.Results:Based on the results of pathological diagnosis, in diagnosing TI-RADS 4 thyroid nodules, the sensitivity, specificity and accuracy were 77.61%, 70.97% and 75.51% for contrast-enhanced ultrasound, respectively; 80.60%, 74.19%, and 78.57% for ultrasound-guided fine-needle aspiration biopsy, respectively; 79.10%, 96.77%, and 84.69% for the BRAF gene test, respectively; and 98.51%, 70.97% and 89.80% for the combined diagnosis, respectively. The AUC was 0.790 for contrast-enhanced ultrasound, and 0.774 for ultrasound-guided fine-needle aspiration biopsy, 0.799 for BRAF genetic testing, and 0.847 for combined testing. The diagnostic value of combined diagnosis was significantly higher than other diagnostic methods ( P<0.05). Conclusions:Contrast-enhanced ultrasound combined with fine-needle aspiration biopsy and BRAF gene detection is valuable for the diagnosis of TI-RADS 4 class thyroid nodules and improves the preperative diagnosis.

Cardio / 中南大学学报(医学版)

Cardio-facio-cutaneous (CFC) syndrome is an extremely rare autosomal dominant genetic disease due to BRAF and other gene mutations. The main characteristics of the patients are craniofacial deformities, cardiac malformations, skin abnormalities, delay of language and motor development, gastrointestinal dysfunction, intellectual disability, and epilepsy. In this case, the child has a typical CFC syndrome face and developmental delay. The gene results of the second-generation sequencing technology showed that there was a mutation site c.1741A>G (p. Asn581Asp) (heterozygous) in exon 14 of the BRAF (NM_004333.5) gene. The mutation was not observed in the child's parents. The above-mentioned mutation may be a de novo mutation. There is no effective therapy for this disease so far.

Logic and clinical significance of the new WHO classification of ameloblastoma in 2017 / 口腔疾病防治

@#A substantial revision of the classification of ameloblastoma was made by the World Health Organization (WHO) in the fourth edition of the Classification of Head and Neck Tumors in 2017, which was based on the review and summary of much clinical research data and prospective evaluation of the latest results of genetic research. The new classification is simpler and more practical. It retains two subtypes, the unicystic type and extraosseous/peripheral type, classifies the remaining types as ameloblastoma (classic), defines metastatic ameloblastoma as a benign tumor and simplifies the classification of ameloblastic carcinoma, which has important guiding significance for clinical diagnosis and treatment. Moreover, the new classification included the latest advances in the genetic research on ameloblastoma, demonstrating that the BRAF gene mutation was found in approximately 60% of ameloblastoma cases. The classification provides a new concept and direction for studying the pathogenesis of ameloblastoma, and BRAF-targeted therapy may be an emerging therapy for some ameloblastoma patients with multiple recurrence or surgical contraindications. This article analyzes the intrinsic logic of these changes via a review of the relevant literature and combination of clinical experiences to better understand the new classification. In 2017, the WHO′s new classification of ameloblastoma summarized the experience and achievements in histopathology and clinical treatment of ameloblastoma in the prior 10 years, indicating that BRAF-targeted treatment may bring new treatment options and hope for patients with recurrent or inoperable ameloblastoma.

Correlation of papillary thyroid carcinoma different subtypes BRAF mutation and ultrasonic characteristics / 中华超声影像学杂志

Objective To investigate the correlation of BRAF mutation frequency with cervical lymph node metastasis ,and to compare the ultrasonic characteristics in patients with BRAF mutation in papillary thyroid carcinoma ( PTC) different subtypes . Methods The tumor samples were collected from 139 PTC patients who underwent thyroidectomy . And they were classified by histological subtype into 3 groups:classic variant of papillary thyroid carcinoma (CVPTC) group( 34 cases) ,follicular variant of papillary thyroidcarcinoma (FVPTC) group(36 cases) ,tall cell variant (TCV) group(69 cases) . The BRAF mutation frequency and the correlation with cervical lymph node metastasis among 3 groups were analyzed , then the ultrasonic characteristics with BRAF mutation in PTC different subtypes were compared . Results①The frequency of BRAF mutation was statistically significant different in different subtypes( χ2 =6 .390 , P =0 .041) ,and the frequency in TCV was 86 .9％ . There was also a statistical difference between BRAF mutation frequency and cervical lymph node metastasis among three subtypes ( χ2 = 13 .106 , P =0 .041) .②There was no statistically significant difference among the three groups in nodule number ,echo level , internal structure ,boundary ,crossbar ,morphology and acoustic halo of patients with BRAF mutation ( P >0 .05) . ③ A single factor analysis was performed for the ultrasonographic characteristics of patients with BRAF mutation ,and there were significant statistical differences among the 3 groups in calcification type (χ2 = 21 .7 , P = 0 .001 ) and close to the envelope (χ2 = 7 .726 , P = 0 .021 ) . ④ Multivariate logistic regression showed that BRAF mutation was an independent influence factor affecting the calcification type of different histological subtypes in PTC patients.Conclusions ①BRAF mutation is correlated with cervical lymph node metastasis in different PTC subtypes . ② BRAF mutation is an independent influence factor affecting the morphology type of different calcification subtypes in PTC . The CVPTC group is mainly microcalcification ,and the TCV group is mainly macrocalcification .

BRAF/ KRAS and PIK3CA mutation characteristics and clinical significance in primary small cell lung caner / 中华放射肿瘤学杂志

Objective To detect the frequency of BRAF/ KRAS and PIK3CA mutations in the small cell lung cancer (SCLC) specimens from a large population of Chinese patients and to analyze the gene mutation and clinical characteristics. Methods A total of 557 samples were collected from SCLC patients from 2009 to 2014.BRAF,KRAS,PIK3CA,NRAS and MEK1 gene mutations were detected by the dideoxy sequencing. Chi-square test was adopted to analyze the correlation between clinical factors and gene mutation. Kaplan-Meier method was utilized for survival analysis. Cox model was used for multivariate prognostic analysis. Results BRAF mutations were detected in 13 out of 557 specimens. The mutation types included V600E (n= 5) ,V600A (n= 2) ,V600M (n= 1) ,D594G (n= 1),G464E (n= 1),K601R (n= 2) and S605N (n= 1).KRAS mutation was detected in 6 cases including G12C (n= 3),G12A (n= 1),G12D (n=1) andG13D (n= 1).PIK3CA mutation was observed in 4 samples including E545G (n= 2) and H1047R (n= 2).Besides,NRAS mutation (Q61R) was detected in 1 case and MEK1 mutation (D61Y) was noted in 1 case. These gene mutations were not significantly correlated with the age, gender, smoking status and clinical staging of the patients. Univariate survival analysis demonstrated the median survival time of patients with gene mutation was (10.30±0. 751) months (95％CI:8. 829-11. 771 months),significantly shorter than (12.80±0. 543) months (95％CI:11. 736-13. 864 months) of their counterparts without gene mutation (P=0. 011). Conclusions BRAF/ KRAS and PIK3CA gene mutation is detected in a small proportion of SCLC patients. These gene mutations are not significantly correlated with the clinical characteristics. Univariate survival analysis demonstrates that negative these gene mutations are negatively correlated with the clinical prognosis of SCLC patients.

Research advances in the v-raf murine sarcoma viral oncogene homolog gene mutation in ameloblastoma / 中华口腔医学杂志

V-raf murine sarcoma viral oncogene homolog B1 (BRAF) is a pro-oncogene, which is one member of the RAF family. Mutated BRAF is found in approximately 8% of human tumors. BRAF gene mutations lead to continuous activation of the mitogen-activatd protein kinase (MAPK) pathway, which resulting in abnormal cell proliferation and tumorigenesis. In recent years, recurrent MAPK signaling mutations were identified in ameloblastoma, among which BRAF-V600E is the most prominent type. This provides new strategies for the targeted treatment of ameloblastoma. This paper reviewed the latest advances in BRAF gene mutation associated with ameloblastoma and its potential clinical significance.

Progress of BRAF gene mutation in children with langerhans cell histiocytosis / 国际儿科学杂志

Langerhans cell histiocytosis (LCH) is a rare disease characterized by clonal proliferation and aggregation of LCH cells(Langerhans cells,LCs),and there has been controversy about its pathogenesis.In recent years,BRAF mutations have been detected repeatedly in LCH patients,suggesting that the BRAF gene mutation may drive the occurrence and development of LCH,and may be associated with clinical chemotherapy and prognosis.These findings not only provide evidence for LCH as a tumor disease,but also lay a molecular genetic basis for the development of targeted therapy for LCH.

Correlation of RAS mutations and microsatellite instability with clinico-pathological features and prognosis of patients with stage Ⅲ colorectal cancer / 中国病理生理杂志

AIM:To investigate the status of RAS/BRAF mutations and microsatellite instability ( MSI ) and their associations with clinicopathological features and prognosis of the patients with stage Ⅲ colorectal cancer ( CRC ) . METHODS:The surgical patients with stage ⅢCRC (n=281) were followed up.The mutations of RAS/BRAF were ex-amined by PCR amplification-Sanger sequencing , and MSI status was detected using immunohistochemistry ( IHC) in their archival paraffin-embedded tissue specimens .The relationships of the status with the clinicopathological features and prog-nosis of the patients were statistically analyzed .RESULTS: Among 281 patients, the mutations of RAS/BRAF were ob-served in 136 cases (48.4%), including 116 cases (41.3%) of KRAS mutations.RAS/BRAF mutations were highly cor-related with the level of carcino-embryonic antigen (P<0.05).Moreover, 18 cases (6.4%) of MSI-high (MSI-H) pa-tients were determined by IHC, and MSI-H status was more common in N2b patients (P<0.05).Correlation study found that the mutation rate of BRAF was higher in MSI-H tumors than that in MSI-low ( MSI-L)/microsatellite stability ( MSS) counterparts (P<0.01), although no association between KRAS/NRAS mutations and the MSI status was observed .The prognosis in the patients with wild-type RAS/BRAF or MSI-H was better than the patients with any mutation or MSI-L/MSS (P<0.01).CONCLUSION:Mutant RAS/BRAF and MSI may serve as fairly good indicators for prognosis of the patients with stage Ⅲ CRC.

Correlation of RAS mutations and microsatellite instability with clinico-pathological features and prognosis of patients with stage Ⅲ colorectal cancer / 中国病理生理杂志

AIM:To investigate the status of RAS/BRAF mutations and microsatellite instability ( MSI ) and their associations with clinicopathological features and prognosis of the patients with stage Ⅲ colorectal cancer ( CRC ) . METHODS:The surgical patients with stage ⅢCRC (n=281) were followed up.The mutations of RAS/BRAF were ex-amined by PCR amplification-Sanger sequencing , and MSI status was detected using immunohistochemistry ( IHC) in their archival paraffin-embedded tissue specimens .The relationships of the status with the clinicopathological features and prog-nosis of the patients were statistically analyzed .RESULTS: Among 281 patients, the mutations of RAS/BRAF were ob-served in 136 cases (48.4%), including 116 cases (41.3%) of KRAS mutations.RAS/BRAF mutations were highly cor-related with the level of carcino-embryonic antigen (P<0.05).Moreover, 18 cases (6.4%) of MSI-high (MSI-H) pa-tients were determined by IHC, and MSI-H status was more common in N2b patients (P<0.05).Correlation study found that the mutation rate of BRAF was higher in MSI-H tumors than that in MSI-low ( MSI-L)/microsatellite stability ( MSS) counterparts (P<0.01), although no association between KRAS/NRAS mutations and the MSI status was observed .The prognosis in the patients with wild-type RAS/BRAF or MSI-H was better than the patients with any mutation or MSI-L/MSS (P<0.01).CONCLUSION:Mutant RAS/BRAF and MSI may serve as fairly good indicators for prognosis of the patients with stage Ⅲ CRC.

Correlation between BRAF gene mutation and papillary thyroid carcinoma / 肿瘤研究与临床

Papillary thyroid carcinoma (PTC) is a common tumor in endocrine system with unclear pathogenesis. The study has shown that the stimulation of cell growth, differentiation and gene mutation factors act together on the thyroid gland cells to transform into tumor cells from normal cells. A variety of oncogenes and tumor suppressor genes are involved in the development of PTC. In recent years, the study has found that BRAF gene mutation has a close relationship with the pathogenesis and prognosis of PTC. Mutated BRAF gene affects the occurrence and development of PTC owing to the regulation by RAS protein kinase and the application of biological function by activating MAPK signaling pathways.

Clinical significance of detection for mutated BRAF gene in thyroid nodules based on amplification refractory mutation system / 中国耳鼻咽喉头颈外科

OBJECTIVE To explore the feasibility of detection for mutated BRAF V600E gene based on amplification refractory mutation system(ARMS),and to evaluate its clinical significance of BRAF V600E gene mutation in thyroid nodules.METHODS The method of ARMS was used to detect BRAF V600E mutation status in 179 patients with PTC and 115 patients with benign lesions.The diagnosis index of BRAF V600E mutation status for identifying the nature of the thyroid nodule was calculated.The potential correlation between BRAF V600E mutation and PTC clinicpathological characteristics was also analyzed.RESULTS Detection of BRAF V600E mutation status in thyroid lesions based on ARMS was feasible and believable.The positive rate of mutated BRAF V600E gene in PTC was 82.68％,whereas the rate in benign lesions was only 1.74％,indicating statistical differences between the two groups(x2=183.568,P＜0.01).The diagnostic sensitivity of BRAF V600E mutation was 82.68％,specificity was 98.26％,accuracy was 88.76％,and Youden index was 0.8094.There was no associations between the BRAF V600E mutation status and PTC clinicpathological characteristics(eg.gender,age,tumor size,numbers of lesions,bilateral lesions,extrathyroidal extension and lymph node metastasis).CONCLUSION Detection of BRAF V600E mutation based on ARMS has higher sensitivity and specificity in distinguishing PTC from benign lesions,indicating BRAF V600E gene is an ideal marker of PTC for clinical early diagnosis.

Melanoma lentiginoso acral en una mujer suazí / Acral lentiginous melanoma in a suazi woman

Se presenta el caso clínico de una fémina de 58 años de edad, de raza negra, quien acudió a la consulta de Cirugía del Hospital Gubernamental de Mbabane en Suazilandia por presentar una lesión pigmentada y ulcerada en el talón del pie derecho, donde se le practicó una biopsia por escisión cuyo resultado fue un melanoma lentiginoso acral invasivo. Posteriormente fue evaluada en la consulta de Oncología y luego de realizarle los exámenes complementarios necesarios, la neoplasia se clasificó en estadio IIC. La paciente fue remitida a Sudáfrica para recibir tratamiento con citosinas inmunomoduladoras, factor estimulante de colonias de granulocitos y macrófagos o inhibidores del gen BRAF.

The case report of a 58 years black woman is presented. She went to the Surgery Service of Mbabane Government Hospital in Suaziland due to a pigmented and ulcerated injury in her right foot heel, where she had an excisional biopsy whose result was an invasive acral lentiginous melanoma. Later on she was evaluated in the Oncology Service and after carrying out the necessary complementary tests, the neoplasm was classified in stage IIC. The patient was referred to South Africa to receive treatment with immunomodulatory cytokines, stimulating factor of granulocytes and macrophages colonies or BRAF gene inhibitors.

BRAF and targeted treatment for non-small cell lung cancer / 国际肿瘤学杂志

B-Raf kinase (BRAF) gene is a driver mutation,and is an effective target in the treatment of non-small cell lung cancer (NSCLC).Studies have shown that BRAF inhibitors are effective for treatment of NSCLC with BRAF mutant.It is important to understand the clinicopathologic features and the research progress of BRAF inhibitors for the individual treatment of NSCLC.

Comparison of real-time PCR method with Sanger sequencing for detection of BRAF muta-tion in papillary thyroid carcinoma / 临床与实验病理学杂志

Purpose To investigate the positive rate and concordance rate of BRAF mutation in papillary thyroid carcinoma detected by real-time PCR method and Sanger sequencing. Methods 312 papillary thyroid carcinomas patients were enrolled in this study. Real-time PCR method and Sanger sequencing were performed to detect BRAF gene mutations. The frequency of BRAF mutation and the concordance of two methods were analyzed. Results BRAF mutation was detected in 65. 4% (204/312) and 63. 8% (199/312) of 312 papillary thyroid carcinoma samples by using real-time PCR method and Sanger sequencing, respectively. There was no significant correlation between BRAF gene mutations and patients’ gender. There was significant correlation between BRAF gene mutations and patients’ age. The overall concordance between real-time PCR method and Sanger sequencing for BRAF mutation detection was 98. 4%. Conclusion Real-time PCR method provides an effective method in BRAF gene mutation detection.

Comparison of BRAF mutation detection in patients with papillary thyroid microcarcinoma by ARMS and direct sequencing / 中国免疫学杂志

Objective:To investigate the sensitivity and the specificity of scorpions amplification refractory mutation system ( ARMS) in comparing with that of direct DNA sequencing in the detection of BRAF gene mutations in patients with papillary thyroid microcarcinoma.Methods:Direct sequencing and ARMS were used simultaneously to detect BRAF mutation status in 56 patients with PTMC.Results:BRAF mutations were identified in 46 cases with a mutation rate of 82.9%by ARMS,while in 18 cases with a mutation rate of 32.1%by direct sequencing.Besides,the sensitivity of ARMS was 100%and that of direct sequencing was 39.1%.There were significant differences of both mutation rate and sensitivity between two methods ( P<0.01 ).Conclusion: Compared to direct sequencing,ARMS gains a higher sensitivity in the detection of BRAF mutations in samples with tiny lesions.

Expression of Cyclin D1 and BRAF (V600 E) mutation in papillary thyroid carcinoma and their correlation with clinicopathologic features / 临床与实验病理学杂志

Purpose To investigate the correlation of the expression of Cyclin D1 and BRAF ( V600E) mutation in papillary thyroid carcinoma ( PTC) and their clinical significance, and to analyze their role in the pathogenesis of PTC. Methods The expression of CK34βE12 and MC protein was detected by immunohistochemical EnVision method in 52 cases of PTC and 52 cases of thyroid benign lesions (25 cases of nodular goiters, 15 cases of Hashimoto’s thyroiditis, 12 cases of follicular adenoma), and the mutation status of BRAF gene protein in above specimens were detected by polymerase chain reaction and DNA sequencing. Expression of Cyclin D1 and BRAF (V600E) mutation in papillary thyroid carcinoma and their correlation clinicopathologic features were also analyzed. Results The positive rate of Cyclin D1 in 88 cases of PTC was 84. 6%,and the immunoreactivity score (4. 6 ± 2. 4), compared with thyroid be-nign lesions (1. 3 ± 1. 6), was statistically significant difference (t=8. 525, P<0. 01). Expression of Cyclin D1 was closely related with lymph node metastasis, capsular invasion, tumor stage Ⅲ+Ⅳ and BRAF (V600E) mutation (P<0. 05), but not related with age, gender, number and diameter of tumor nodules. In 52 cases of PTC BRAF gene mutation rate was 63. 5%, but in thyroid benign lesions mutation rate was 0, with significant difference (P<0. 01). The BRAF mutation is closely related with lymph node metastasis, capsular invasion and tumor stage Ⅲ+Ⅳ(P<0. 05), but not related with age, gender, number and diameter of tumor nodules. The expression of Cyclin D1 in BRAF (V600E) mutation group was stronger, the positive expression rate was significantly higher than that of wild group (100% vs 57. 9%), and expression of the mean score was higher than that of the wild group (5. 7 ± 1. 6 vs 4. 0 ± 2. 5);the comparison between the two groups showed statistical difference (t=2. 652, P<0. 05). Conclusions The expression of Cyclin D1 and BRAF (V600E) mutation are positively related, which are also closely related with the occurrence and development of PTC, and can be used as an index to predict the invasion and metastasis in PTC.

Expression of pyruvate kinase isoform M2 and its relationship with BRAF mutation in papillary thyroid carcinoma / 肿瘤

Objective: To explore the relationship between PKM2 (M2 isoform of pyruvate kinase) expression and the clinicopathological features and BRAF (B-type Raf kinase) gene mutation in human PTC (papillary thyroid carcinoma). Methods: The expressions of PKM2 protein in 82 human PTC tissue samples, 40 human normal thyroid tissue samples and 20 human nodular goiter tissue samples were detected by immunohistochemistry. The BRAF gene mutation status in PTC tissues was examined using PCR (polymerase chain reaction) amplication and direct sequencing. Results: The expression level of PKM2 protein varied in 91.5% (75 of 82) of PTC tissues. Otherwise, there was no expression of PKM2 in normal thyroid tissues and nodular goiter tissues. The expression of PKM2 was associated with T staging and lymphatic metastasis (P < 0.05). The positive rates of PKM2 expression in 46 PTC tissue samples carrying a BRAF mutation and 36 PTC tissue samples carrying wild type BRAF were 100.0% (46/46) and 86.1 % (31/36), respectively (P < 0.05). Conclusion: This study demonstrated that the over-expression of PKM2 is associated with the aggressiveness of PTC and BRAF gene mutation status. Copyright © 2013 by TUMOR.

Effects of BRAFV600E mutation on the invasion capacity of human melanoma cells / 中华皮肤科杂志

Objective To investigate the effect of BRAFV600E mutation on the invasion capacity of a human melanoma cell line, A375. Methods Plasmids containing short hairpin RNAs (shRNA) specific for BRAF gene were prepared in previous study, and used to transfect A375 cells. Those cells transfected with negative plasmid and untransfected cells served as the controls. Transwell chambers were used to examine the invasion ability of melanoma cells in vitro. RT-PCR and Western blotting were performed to detect the mRNA and protein expressions of matrix metalloproteinase 2 (MMP-2) and vascular endothelial growth factor (VEGF), respectively, before and after the transfection. The activity of MMP-2 was also studied with sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Results Compared with the negative control, the specific shRNA decreased the mRNA and protein expressions of MMP-2 by 35% and 85%, respectively, and those of VEGF by 45% and 14%, respectively. Additionally, the number of cells invading through Matrigel chambers reduced by 69% in those cells transfected with the positive plasmid. Conclusions The mutant BRAFV600E has the potential to enhance the invasion capacity of melanoma cells, whereas specific shRNA could suppress the increase in metastasis capacity likely by inhibiting the production of VEGF and MMP.