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摘要:
目的 探讨1例原发性肾性糖尿(primary renal glucosuria, PRG)患者的临床表现及其分子生物学基础。 方法 详细收集患者的临床资料、生化检查及影像学检查结果, 抽取外周静脉血, 提取基因组DNA, 聚合酶链反应(polymerase chain reaction, PCR)扩增SLC5A2基因的14个外显子及其与内含子的交界区, 测序确定突变情况。 结果 患者临床表现、实验室检查和影像学检查符合PRG诊断。基因突变分析显示, 患者SLC5A2基因cDNA序列的第877位腺嘌呤A突变为胸腺嘧啶G(c.877 A>G), 造成第293位氨基酸由丝氨酸改变为半胱氨酸(p.Ser293Cys), 该突变位于SLC5A2的第7外显子。蛋白序列的保守性分析和突变蛋白的功能分析均表明p.Ser293Cys为致病性突变。 结论 通过SLC5A2基因突变分析, 从分子遗传学方面证实患者PRG的诊断。临床上血糖正常的患者出现尿糖阳性且无其他近端肾小管功能障碍表现的患者应该考虑到该疾病可能, 基因分析有助于确诊。 -
关键词:
- 原发性肾性糖尿 /
- 家族性肾性糖尿 /
- SLC5A2基因 /
- 钠-葡萄糖共转运蛋白2 /
- 基因突变
Abstract:Objective To analyze the clinical manifestations and molecular basis of a patient with primary renal glucosuria (PRG). Methods Clinical features, laboratory data, and imaging results were collected. Genomic DNA was extracted from leukocytes of peripheral blood of the patient. Fourteen exons of the PRG gene and their boundaries with introns were amplified by polymerase chain reaction (PCR). The mutations of the SLC5A2 gene were identified by direct sequencing. Results PRG was diagnosed on the basis of comprehensive consideration of clinical presentations, laboratory test results, and imaging findings. Gene mutation test revealed a nucleotide substitution of guanine for adenine at the position 877 of cDNA sequence of SLC5A2 gene (c.877 A>G), which caused a missense mutation of serine to cysteine at codon 293 (p.Ser293Cys). It occurred at the 7th exon of SLC5A2. Loci conservation analysis and functional prediction of missense mutation of SLC5A2 protein revealed that p.Ser293Cys was a pathogenic mutation. Conclusions SLC5A2 gene mutation analysis confirms the diagnosis of PRG in this patient from the aspect of molecular genetics.It is important to suspect PRG in patients with renal glucosuria and normal blood glucose in the absence of other manifestations ofproximal renal tubular dysfunction.Genetic analysis of SLC5A2 may be helpful to confirm the diagnosis. -
Key words:
- primary renal glucosuria /
- familial renal glucosuria /
- SLC5A2 gene /
- SGLT2 protein /
- gene mutation
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图 2 不同种属的SGLT2蛋白多序列比对图
框内为SGLT2蛋白第293位的丝氨酸，该氨基酸在人、大鼠、小鼠、兔、牛、大猩猩、马、荷兰猪、恒河猴和狗的SGLT2蛋白序列中高度保守
表 1 SLC5A2基因扩增引物
外显子 上游引物序列 下游引物序列 PCR产物长度(bp) 退火温度(℃) 延伸时间(s) 1 GGCAGATCCTGGGGAGAA CCCTGAGAACTTCCAAAGACC 295 59 20 2~3 CGATCAGGGAAACTTGGCTC CTTCCTGGCTAGATGTATGACC 536 60 40 4~5 TTCCCAGGGCCACTTGCTTG AGTTCCTTCTCCTGCTTGGAC 782 60 50 6~7 CCCGAGAACAGGCTATCGTT CTCAGACTGCGCCTTAGACC 683 62 40 8~9 GACAGAAGGCTCCATCTACTCCA GCATGACCGCCAGCATGAG 754 63 50 10~12 GAAGCTCATGCCCAACGGTA CCATTCCCAACCCTCAGTCG 976 63 60 13~14 AGTTGGCATGAGTTAAGCCT GGGAAGTGACTGCCAATCAGA 749 61 50 
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[1] Santer R, Kinner M, Lassen CL, et al. Molecular analysis of the SGLT2 gene in patients with renal glucosuria[J]. J Am Soc Nephrol, 2003, 14:2873-2882. doi: 10.1097/01.ASN.0000092790.89332.D2 [2] Lee H, Han KH, Park HW, et al. Familial renal glucosuria:a clinicogenetic study of 23 additional cases[J]. Pediatr Nephrol, 2012, 27:1091-1095. doi: 10.1007/s00467-012-2109-9 [3] Sim NL, Kumar P, Hu J, et al. SIFT web server:predicting effects of amino acid substitutions on proteins[J].Nucleic Acids Res, 2012, 40:W452-W457. doi: 10.1093/nar/gks539 [4] Adzhubei IA, Schmidt S, Peshkin L, et al. A method and server for predicting damaging missense mutations[J]. Nat Methods, 2010, 7:248-249. doi: 10.1038/nmeth0410-248 [5] Schwarz JM, Cooper DN, Schuelke M, et al. Mutation Taster 2:mutation prediction for the deep-sequencing age[J]. Nat Methods, 2014, 11:361-362. doi: 10.1038/nmeth.2890 [6] Calado J, Sznajer Y, Metzger D, et al. Twenty-one additional cases of familial renal glucosuria:absence of genetic heterogeneity, high prevalence of private mutations and further evidence of volume depletion[J]. Nephrol Dial Transplant, 2008, 23:3874-3879. doi: 10.1093/ndt/gfn386 [7] Santer R, Calado J. Familial renal glucosuria and SGLT2:from a mendelian trait to a therapeutic target[J]. Clin J Am Soc Nephrol, 2010, 5:133-141. doi: 10.2215/CJN.04010609 [8] Yu L, Hou P, Lv JC, et al. Novel SLC5A2 variants contribute to renal glucosuria in Chinese families:abnormal expression and dysfunction of variant SLC5A2[J]. Hum Mutat, 2015, 36:79-86. doi: 10.1002/humu.22714 [9] Yu L, Hou P, Lv JC, et al. A novel sodium-glucose co-transporter 2 gene (SGLT2) mutation contributes to the abnormal expression of SGLT2 in renal tissues in familial renal glucosuria[J]. Int Urol Nephrol, 2014, 46:2237-2238. doi: 10.1007/s11255-014-0755-5 [10] Lee YW. Clinical and genetic analysis in a patient with primary renal glucosuria:Identification of a novel mutation in the SLC5A2 gene[J]. Exp Ther Med, 2013, 6:1532-1534. doi: 10.3892/etm.2013.1326 [11] Yu L, Lv JC, Zhou XJ, et al. Abnormal expression and dysfunction of novel SGLT2 mutations identified in familial renal glucosuria patients[J]. Hum Genet, 2011, 129:335-344. doi: 10.1007/s00439-010-0927-z -
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