Expression Of BRAF And Its Significance With Malignant Melanoma Of Xinjiang Region

Objective: To examed the difference in BRAF gene expression between malignant melanoma(MM),pigment nevi and normal skin. Analyzed the relationship among BRAF gene expression, malignantmelanoma clinical phenotype, so explored its relationship with the occurrence and development ofmalignant melanoma, in order to find a new indicator for disease prediction and prognosis. The method ofextraction of RNA from formalin fixed and paraffin embedded(FFPE) of MM tissue for reversetranscription and polymerase chain reaction(RT-PCR)were established. The feasibility of this method wereinvestigated.Methods：This study involved formalin-fixation and paraffin-embedding tissue samples from74casesmalignant melanoma patients,35cases pigment nevi and46cases Normal skin tissue in People’s Hospitalof Xinjiang Uygur Autonomous Region. Clinical data were collected and analyzed. The expression ofBRAF gene were detected by SYBR Green real time PCR in74cases of malignant melanoma and35casespigment nevi and46cases Normal skin tissue cases. Each sample were calculated and analysized on2-ΔCt.RNA were extracted from FFPE sections and examed by agarose gel electrophoresis and ultravioletspectrophotometry. Feasibility of RT-PCR is made of the establishment of solubility curves andamplification plot. The relationship between BRAF expression and clinical phenotype were analyzed by ttest or one-way ANOVA test according to the SPSS17.0Software.Results：34of74cases in malignant melanoma patients were Uygur races (45.9%),the Han nationalitywere40cases (54.1%); The males were39cases (52.7%), the females were35cases (47.3%), the ratiobetween male and female is1.11:1; The average age is55.6±14.3, the age of11cases is less than or equal45years (14.9%), the age of20cases is between46and59(27.0%), the age of43cases is more than orequal to60years (58.1%); the primary site was located in the skin which were47cases(63.5%),24cases in the mucosa(32.4%), other is3cases(4.1%);33cases of47cases were occurred in theextremities(70.2%).2)Comparison of the expression of BRAF gene among Uygur races and Hannationality malignant melanoma, pigment nevi and normal skin tissue: the expression of BRAF gene inmalignant melanoma was0.377±0.167, in pigment nevi was0.159±0.167and in normal skin tissue was0.134±0.050. The results showed that the expression of BRAF gene in malignant melanoma was highersignificantly than that in pigment nevi and skin normal tissue (p<0.01). The expression of BRAF inmelanoma patients were no significant correlation with Clinical pathology characteristics such as genderage, ethnicity, anatomical site, metastasis of lymph node, ulceration(P>0.05). The MM of extraction ofRNA from formalin fixed and paraffin embedded degrade at different degree. Concentration and purity ofRNA fit for the demand of experiment. The result of RT-PCR have shown that dissolved curve with asingle-peak. The target gene and reference gene to amplification efficiency are mostly identical.Conclusion:1）The incidence of melanoma mainly in the elderly, with small gender differences inXinjiang region, acral is predilection sites.2）The higher expression of BRAF gene may involve in theoccurrence and development of melanoma and play an important role in the melanoma.