多发性骨髓瘤骨髓RAS基因突变检测分析

Analysis of RAS gene mutations in patients with multiple myeloma

目的:检测NRAS和KRAS基因在多发性骨髓瘤(MM)患者中的突变类型,探讨其在MM病理发生发展过程中的作用。方法:采用多聚酶链反应和DNA序列测定的方法,对50例MM患者骨髓细胞基因组中的NRAS基因第1和第2外显子以及KRAS基因的外显子进行检测,并与标准参考序列进行对比。结果:50例MM患者中有3例(6%)Ⅲ期男患者发现NRAS基因突变,分别为第1外显子编码区第34位核苷酸的杂合突变c.34G>A,导致第12密码子发生GCT>AGT错义突变,从而产生氨基酸Gly>Ser(G12S)的改变;c.38G>A杂合突变,导致第13密码子发生GGT>GAT,Gly>Asp(G13D)的错义突变;c.182A>T杂合突变,导致位于第2外显子中的第61密码子发生CAA>CTA,Gln>Leu(Q61L)的错义突变。未检测到KRAS基因外显子区的突变。结论:NRAS突变可能与MM的病理进程相关,NRAS G12S、G13D和Q61L突变在MM发生和发展中的功能和作用还有待进一步的深入研究。

OBJECTIVE：To analyze NRAS and KRAS gene mutations in patients with multiple myeloma （MM） and evaluate their clinical relevance to the process of MM. METHODS：By using polymerase chain reaction and DNA sequencing techniques, the exon 1,exon 2 of NRAS gene and the all exons of KRAS gene were analyzed in bone marrow-derived genomic DNA of fifty MM patients,and the sequences of patients＇ samples were aligned with standard reference se- quences. RESULTS：In 50 MM patients, three NRAS gene mutations were identified in 3 patients （6%） with phase []] stage of MM,which included a c. 34 G〉A mutant heterozygote in exon 1 that resulted GCT〉AGT missense mutation in Codon 12 and Gly〉Ser （G12S） amino acid change; a c. 38G〉A mutant heterozygote in exon 1 that resulted GCT〉GAT missense mutation in Codon 13 and Gly〉Asp （G13D） amino acid change; and a c. 182 A〉T mutant heterozygote in exon 2 that resulted CAA〉CTA missense mutation in Codon 61 and Gln〉Leu （Q61L） amino acid change,respectively. KRAS gene mutations were not found in exon regions. CONCLUSION：The NRAS mutations may relevant to pathological process of MM, the functional roles of the NRAS mutations G12S,G13D and Q61L played in MM development still needs further investigation.