白血病患者线粒体基因组D-loop区基因不稳定性研究
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摘要
目的:探讨白血病患者线粒体基因组D-loop区基因的不稳定性。方法:采用PCR扩增24例患者D-loop区HV-1和HV-2两个高变区并测序,其结果与剑桥标准序列(rCRS)和mtDB数据库进行比对,分析其突变情况。采用SPSS 22.0统计学软件对数据进行分析。结果:23例患者D-loop区发生突变,突变率95.83%(23/24);共检测到82个基因突变,其中HV-1区47个(57.32%),HV-2区35个(42.68%)。急性髓系白血病患者中未治疗组(UT)与治疗组(T)对比:突变率分别为(2.37±0.82)×10~(-3)和(4.76±2.45)×10~(-3),其差异有统计学意义(P<0.01)。急性髓系白血病治疗组中部分缓解组(PR)和完全缓解组(CR)突变率比较显示,突变率分别为(5.10±2.56)×10~(-3)和(4.51±2.51)×10~(-3),其差异无统计学意义(P>0.05)。M3组间比较显示,未治疗组(UT)、部分缓解组(PR)和完全缓解组(CR)突变率分别为(2.55±0.63)×10~(-3),(5.37±3.41)×10~(-3)和(3.71±1.65)×10~(-3),其差异无统计学意义(P>0.05)。结论:D-loop区中存在较高的突变率和多种突变类型,具有较强的不稳定性;化疗可能加剧D-loop区基因的不稳定性。
Objective: To study the instability of mitochondrial DNA(mt DNA) D-loop region genes in patients with Leukemia. Methods: The HV-1 and HV-2 regions of D-loop region in 24 patients with leukemia were amplificated and sequenced, then their results were compared with revised Cambridge reference sequence(rCRS) and Databank mtDB. The mutation rate was detected by SPSS 22.0 statistics software. Results: The total mutation rate in patients was95.83%(23/24), the detection showed 82 mutated genes, out of which 47(57.32%) mutated genes located in HV-1 region,35(42.68%) mutated genes in HV-2 region. The comparison showed that the mutation rate in untreated(UT) group and treated(T) group of AML patients was(2.37 ± 0.82) × 10-3 and(4.76 ± 2.45) × 10-3 respectively(P<0.01),the mutation rate in PR and CR groups of treated AML patients was(5.10 ± 2.56) × 10~(-3) and(4.51 ± 2.51) × 10~(-3) respectively(P < 0.05),the comparison among M3 group showed that the mutation rates in UT,PR and CR groups were(2.55 ± 0.63) × 10~(-3),(5.37 ± 3.41) × 10~(-3) and(3.71 ± 1.65) × 10-3 respectively(P>0.05), Conclusion: The more high mutation rate and many kinds of mutation types exist in D-loop region, suggesting that the genes in D-loop region display the more strong instability, the chemotherapy may aggravate the instability of genes in D-loop region.
Objective: To study the instability of mitochondrial DNA(mt DNA) D-loop region genes in patients with Leukemia. Methods: The HV-1 and HV-2 regions of D-loop region in 24 patients with leukemia were amplificated and sequenced, then their results were compared with revised Cambridge reference sequence(rCRS) and Databank mtDB. The mutation rate was detected by SPSS 22.0 statistics software. Results: The total mutation rate in patients was95.83%(23/24), the detection showed 82 mutated genes, out of which 47(57.32%) mutated genes located in HV-1 region,35(42.68%) mutated genes in HV-2 region. The comparison showed that the mutation rate in untreated(UT) group and treated(T) group of AML patients was(2.37 ± 0.82) × 10-3 and(4.76 ± 2.45) × 10-3 respectively(P<0.01),the mutation rate in PR and CR groups of treated AML patients was(5.10 ± 2.56) × 10~(-3) and(4.51 ± 2.51) × 10~(-3) respectively(P < 0.05),the comparison among M3 group showed that the mutation rates in UT,PR and CR groups were(2.55 ± 0.63) × 10~(-3),(5.37 ± 3.41) × 10~(-3) and(3.71 ± 1.65) × 10-3 respectively(P>0.05), Conclusion: The more high mutation rate and many kinds of mutation types exist in D-loop region, suggesting that the genes in D-loop region display the more strong instability, the chemotherapy may aggravate the instability of genes in D-loop region.
引文
1刘晓红,朱明华.线粒体基因组与肿瘤的相关性研究进展.中国癌症杂志,2003;13(6):587-590.
2 Penta JS,Johnson FM,Wachsmsn JT,et a1.Mitoehondrial DNA in human malignancy.Murat Res,2001;488(2):119-133.
3陈明,周永安.急性白血病及淋巴瘤线粒体基因组D-loop区不稳定性研究.硕士学位论文,2010.
4陈明,周永安,马云霞,等.急性白血病线粒体D-loop区微卫星D310和D16184不稳定性研究.白血病·淋巴瘤,2011;20(1):15-17.
5 Shadel GS,Clayton DA.Mitochondrial DNA maintenance in vertebrates.Annu Rev Biochem,1997;66(1):409-435.
6 Binachi NO,Binachi MS,Richard SM.Mitochondrial genome instability in human cancers.Mutat Res,2001;488(1):9-23.
7 Taanman JW.The mitochondrial genome:structure,transcription,tr anslation and replication.Biochim Biophys Acta,1999;1410(2):103-123.
8 Schon EA.Mitochondrial genetics and disease.Trends Biochem Sci,2000;25(11):555-560.
9 Abnet CC,Huppi K,Carrera A,et a1.Control region muations and the common delection are frequent in the mitochondrial DNA of patients with esophagcal sequamous cell carcinoma.BMC Cancer,2004;4(1):30.
10 Habano W,Sugai T,Nakamura ST,et a1.Microsatellite instability and mutation of mitochondrial and nuclearDNA in gastric carcinoma.Gastroenterology,2000;118(5):835-841.
11 Rosson D,Keshgegian AA.Frequent mutations in the mitochondrial control region DNA inbreast tissue.Cancer Lett,2004;215(1):89-94.
12 Poetsch M,Petersmann A,Lignitz E,et a1.Relationship between mitochondrial DNA instability,mitochondrial DNA large deletions,and nuclear microsatellite instability in head and necksquamous cell carcinomas.Diagn Mol Pathol,2004;13(1):26-32.
13 Hiyama T,Tanaka S,Shima H,et a1.Somatic mutation in mitochondrial DNA and nuclear microsatellite instability in gastriccancer.Oncol Rep,2003;10(6):1837-1841.
14 Parrella P,Xiao Y,Fiss M,et a1.Detection of mitochondrial DNAmultations in Primary breast cancer and fine-needle aspirates.Cancer Res,2001;61(20):7623-7626.
15 Grist SA,Lu XJ,Morley AA.Mitochondrial mutations in acute leukemia.Leukemia,2004;18(7):1313-1316.
16 Carew JS,Zhou Y,Albitar M,et a1.Mitochondrial DNA mutations in primary leukemia cells after chemotherapy:clinical significance and therapeutic implications.Leukemia,2003:17(8):1437-1447.
17 Meng R,Zhou J,Sui M,et a1.Arsenic trioxide promotes mitochondrial DNA mutation and cell apoptosis in primary APL cells and NB4cell line-.Sci China Life Sci.,2010;53(1):87-93.
18 Sharawat SK,Bakhshi R,Vishnubhatla S,et a1.Mitochondrial D-loop variations in paediatric acute myeloid leukemia:a potential prognostic marker.Br J Haematol,2010;149(3):391-398.
2 Penta JS,Johnson FM,Wachsmsn JT,et a1.Mitoehondrial DNA in human malignancy.Murat Res,2001;488(2):119-133.
3陈明,周永安.急性白血病及淋巴瘤线粒体基因组D-loop区不稳定性研究.硕士学位论文,2010.
4陈明,周永安,马云霞,等.急性白血病线粒体D-loop区微卫星D310和D16184不稳定性研究.白血病·淋巴瘤,2011;20(1):15-17.
5 Shadel GS,Clayton DA.Mitochondrial DNA maintenance in vertebrates.Annu Rev Biochem,1997;66(1):409-435.
6 Binachi NO,Binachi MS,Richard SM.Mitochondrial genome instability in human cancers.Mutat Res,2001;488(1):9-23.
7 Taanman JW.The mitochondrial genome:structure,transcription,tr anslation and replication.Biochim Biophys Acta,1999;1410(2):103-123.
8 Schon EA.Mitochondrial genetics and disease.Trends Biochem Sci,2000;25(11):555-560.
9 Abnet CC,Huppi K,Carrera A,et a1.Control region muations and the common delection are frequent in the mitochondrial DNA of patients with esophagcal sequamous cell carcinoma.BMC Cancer,2004;4(1):30.
10 Habano W,Sugai T,Nakamura ST,et a1.Microsatellite instability and mutation of mitochondrial and nuclearDNA in gastric carcinoma.Gastroenterology,2000;118(5):835-841.
11 Rosson D,Keshgegian AA.Frequent mutations in the mitochondrial control region DNA inbreast tissue.Cancer Lett,2004;215(1):89-94.
12 Poetsch M,Petersmann A,Lignitz E,et a1.Relationship between mitochondrial DNA instability,mitochondrial DNA large deletions,and nuclear microsatellite instability in head and necksquamous cell carcinomas.Diagn Mol Pathol,2004;13(1):26-32.
13 Hiyama T,Tanaka S,Shima H,et a1.Somatic mutation in mitochondrial DNA and nuclear microsatellite instability in gastriccancer.Oncol Rep,2003;10(6):1837-1841.
14 Parrella P,Xiao Y,Fiss M,et a1.Detection of mitochondrial DNAmultations in Primary breast cancer and fine-needle aspirates.Cancer Res,2001;61(20):7623-7626.
15 Grist SA,Lu XJ,Morley AA.Mitochondrial mutations in acute leukemia.Leukemia,2004;18(7):1313-1316.
16 Carew JS,Zhou Y,Albitar M,et a1.Mitochondrial DNA mutations in primary leukemia cells after chemotherapy:clinical significance and therapeutic implications.Leukemia,2003:17(8):1437-1447.
17 Meng R,Zhou J,Sui M,et a1.Arsenic trioxide promotes mitochondrial DNA mutation and cell apoptosis in primary APL cells and NB4cell line-.Sci China Life Sci.,2010;53(1):87-93.
18 Sharawat SK,Bakhshi R,Vishnubhatla S,et a1.Mitochondrial D-loop variations in paediatric acute myeloid leukemia:a potential prognostic marker.Br J Haematol,2010;149(3):391-398.