【作者】 宋云风；

【导师】 张军宁；

【作者基本信息】 苏州大学 ， 肿瘤放射治疗学， 2009， 硕士

【摘要】 目的:通过检测鼻咽癌患者外周血的XRCC1 Codon399单核苷酸多态性,分析各基因型与肿瘤放射敏感性和正常组织放射反应的相关性。方法:经病理学确诊的鼻咽癌患者60例(均为低分化鳞癌),于放疗前抽取静脉血,抽取全血基因组DNA进行PCR扩增,对扩增产物再进行酶连接检测反应扩增,最后于测序仪电泳通过分析软件得出XRCC1 Codon399的基因型。分别于放疗中、放疗结束后及放疗后三个月进行临床疗效评价,比较不同基因型与肿瘤放射敏感性和正常组织急、慢性放射反应的相关性。结果:1.在60例患者中携带XRCC1 Codon399Gln/Gln基因型6例,占10%;Arg/Gln基因型21例,占35%;Arg/Arg基因型33例,占55%。各基因型与鼻咽癌患者的性别、年龄、T分期、N分期、临床分期经统计学分析均无显著性差异(P>0.05)。2.放疗中期(改野时)鼻咽癌肿瘤退缩率:携带XRCC1 Codon399Gln/Gln基因型者为0.899±0.0590,显著高于携带Arg/Gln基因型者0.488±0.0175和Arg/Arg基因型者0.416±0.0189。经统计学分析均有高度显著性差异(P<0.001)。3.放疗结束后鼻咽癌肿瘤退缩率:携带XRCC1 Codon399Gln/Gln基因型者为0.9 98±0.0020,显著高于携带Arg/Gln基因型者0.817±0.0231和携带Arg/Arg基因型者0.794±0.0159。经统计学分析均有显著性差异(P<0.05)。4.放疗后三个月鼻咽癌的疗效:携带XRCC1 Codon399Gln/Gln基因型患者放疗后三个月的有效(CR+PR)率为100%,Arg/Gln基因型的有效(CR+PR)率为100%, Arg/Arg基因型的有效(CR+PR)率为93.9%。经统计学分析:三种基因型之间无显著性差异(P>0.05)。5.临床分期与鼻咽癌放疗后三个月疗效:临床分期Ⅰ、Ⅱ、Ⅲ期、ⅣA有效(CR+PR)率分别为100%、100%、96.2%、85.7%。临床分期Ⅰ、Ⅱ、Ⅲ期的有效率高于ⅣA的有效率,经统计学分析均有显著性差异(P<0.05)。但Ⅰ、Ⅱ、Ⅲ期之间无显著性差异(P>0.05)。6. XRCC1 Codon399单核苷酸多态性与正常组织(皮肤、粘膜、涎腺)的急性放射损伤:所有患者在放疗过程中均发生了不同程度的正常组织急性放射损伤。在皮肤发生1级、2级、3级急性放射损伤中,携带XRCC1 Codon399Gln/Gln基因型患者的发生率均为33.3%,Arg/Gln基因型的发生率分别为52.3%、42.9%、4.8%,Arg/Arg基因型的发生率分别为57.6%、36.3%、6.1%;在粘膜发生1级、2级、3级急性放射损伤中,携带XRCC1 Codon399Gln/Gln基因型患者的发生率分别为33.3%、50%、16.7%,Arg/ Gln基因型的发生率分别为38.1%、52.4%、9.5%,Arg/Arg基因型的发生率分别为48.5%、42.4%、9.1%;在涎腺发生1级、2级急性放射损伤中,携带XRCC1 Codon399Gln/Gln基因型患者的发生率分别为0%、100%,Arg/Gln基因型的发生率分别为33.3%、66.7%,Arg/Arg基因型的发生率分别为36.4%、63.6%。以上结果经统计学分析均无显著性差异(P﹥0.05),即正常组织急性放射损伤的发生与XRCC1 Codon399单核苷酸多态性无明显相关性。7. XRCC1 Codon399单核苷酸多态性与正常组织(皮肤、皮下组织、粘膜、涎腺)的晚期放射损伤:所有患者在放疗结束后均发生了不同程度的正常组织晚期放射损伤。在皮肤发生1级、2级、3晚期放射级损伤中,携带XRCC1 Codon399Gln/Gln基因型患者的发生率分别为33.3%、66.7%、0%,Arg/Gln基因型的发生率分别为52.4%、42.9%、4.7%,Arg/Arg基因型的发生率分别为63.6%、36.4%、0%;在皮下组织发生1级、2级、3级晚期放射损伤中,携带XRCC1 Codon399Gln/Gln基因型患者的发生率分别为33.3%、66.7%%、0%,Arg/Gln基因型的发生率分别为38.1%、57.2%、4.7%,Arg/Arg基因型的发生率分别为42.4%、57.6%、0%。;在粘膜发生1级、2级、3级晚期放射损伤中,携带XRCC1 Codon399Gln/Gln基因型患者的发生率分别为100%、0%、0%,Arg/Gln基因型的发生率分别为80.9%、14.3% 4.7%,Arg/Arg基因型的发生率分别为93.9%、6.1%、0%;在涎腺发生1级、2级、3级晚期放射损伤中,携带XRCC1 Codon399Gln/Gln基因型患者的发生率分别为100%、0%、0%,Arg/Gln基因型的发生率分别为80.9%、14.3%、4.7%,Arg/Arg基因型的发生率分别为93.9%、6.1%、0%。以上结果经统计学分析均无显著性差异(P﹥0.05),即正常组织发生的晚期放射损伤与XRCC1 Codon399单核苷酸多态性无明显相关性。结论:(1)携带XRCC1 Codon399 Gln/Gln基因型的鼻咽癌患者放疗中期及结束后肿瘤退缩率大于其它两种基因型,其基因单核苷酸多态性可以作为预测鼻咽癌放疗中期及结束后疗效的因素。(2)三种基因型的鼻咽癌患者放疗后3个月之间的疗效基本相同。放疗后3个月的疗效主要与鼻咽癌的临床分期有关。(3)XRCC1 Codon399单核苷酸多态性与鼻咽癌患者正常组织急性及晚期放射损伤无明显相关性。更多还原

【Abstract】 Objective: By detecting the single nucleotide polymorphisms of XRCC1 Codon399 of peripheral blood of patients suffering from nasopharyngeal carcinoma(NPC) ,to analyse the relation between single nucleotide polymorphisms of XRCC1 Codon399 and radiation injury in normal tissues.Methods:Phlebotomize 60 cases of patients with nasopharyngeal carcinoma which are all confirmed of low differentiated squamous cell carcinoma by pathology . Extract genome DNA from whole blood cells before radiotherapy. Then perform PCR, and amplify the PCR products by ligase detection reaction, put the LDR products on sequencer to carry out electrophoresis, get the genotype of XRCC1 Codon399, to observe the acute irradiation-induced injury of normal tissue during radiotherapy. To evaluate the clinical effects -in the mid-course、the last phase and three months after radiotherapy, to compare the acute and chronic radiation reactions of normal tissue in the different genotypes.Results:(1) Of all 60 cases, the number of XRCC1 Codon399 Gln/Gln was 6, accounting for 10%. Arg/Gln was 21, accounting for 35%. Arg/Arg was 33, accounting for 55%.There was no significant difference among the three genotypes on age, gender, clinical stage, T stage, N stage (P>0.05).(2) In mid-course of radiotherapy the regression rate of nasopharyngeal cancer with Gln/Gln genotype wa(s0.899±0.0590), obviously higer than Arg/Gln (0.488±0.0175) and Arg/Arg genotype(0.416±0.0189), There were significant differences among the three genotypes (P <0.05).(3) At the end of radiotherapy the regression rate of Gln/Gln genotype was (0.998±0.0020),obviously higer than that of Arg/Gln (0.817±0.0231) and Arg/Arg genotype(0.794±0.0159), there were significant differences among the three genotypes(P <0.05).(4) Three months after radiotherapy, the efficiency of cases (CR + PR) with Gln/Gln was 100%, Arg/Gln was 100%, Arg/Arg was 93.9%. But there was no significant difference among the three genotypes (P>0.05).(5) The relation between clinical stage and the efficiency in three months after radiotherapy. The efficiency of cases (CR + PR) was 100%, 96.2%, 85.7% at clinical stageⅠ,Ⅱ,Ⅲ,ⅣA respectively. The efficiency of Clinical stageⅠ,Ⅱ,Ⅲis higher than clinical stageⅣA. There were highly significant differences (P <0.01). But there was no significant difference among the clinical stageⅠ,Ⅱ,Ⅲ(P>0.05).(6) The relation between single nucleotide polymorphisms of XRCC1 Codon399 and acute radiation injury in normal tissues (skin, mucosa, parotid gland):All cases had suffered from the acute irradiation-induced injury in normal tissues to some extent. The rate of the acute irradiation-induced injury of skin at Grade 1 ,2, 3 was all 33.3% with Gln/Gln genotype,52.3%, 42.9%, 4.8% with Arg / Gln and 57.6%, 36.3%, 6.1% with Arg / Arg genotype respectively. The rate of the acute irradiation-induced injury of mucosa at Grade 1 ,2, 3 was 33.3%, 50.0%, 16.7% with Gln/Gln, 38.1%, 52.4%, 9.5% with Arg/Gln and 48.5% , 42.4%, 9.1% with Arg/Arg genotype respectively. The rate of the acute irradiation-induced injury of parotid gland at Grade 1 ,2 was 0%, 100% with Gln/Gln, 33.3%, 66.7% with Arg/Gln, 36.4%, 63.6% with Arg/Arg genotype, respectively. All results showed there was no significant difference among the three genotypes (P>0.05). (7) The relation between single nucleotide polymorphisms of XRCC1 Codon399 and chronic radiation injury in normal tissues (skin, subcutaneous tissue, mucosa, parotid gland):All cases had suffered from the chronic irradiation-induced injury in normal tissues to some extent. The rate of the chronic irradiation-induced injury of skin at Grade 1 ,2, 3 was 33.3%, 66.7%%, 0% with Gln/Gln, 52.4%,42.9%,4.7% with Arg/Gln, 63.6%,36.4%, 0% with Arg/Arg genotype respectively. The rate of the chronic irradiation-induced injury of subcutaneous tissue at Grade 1 ,2, 3 was 33.3%, 66.7%%, 0% with Gln/Gln 42.4%, 57.6%, 0% with Arg/Gln , 38.1%、57.2%、4.7% with Arg/Arg genotype .The rate of the chronic irradiation-induced injury of mucosa at Grade 1, 2, 3 was 100%, 0%, 0% with Gln/Gln, 93.9%, 6.1%, 0% with Arg / Gln, 80.9%, 14.3%, 4.7% with Arg/Arg genotype respectively. The rate of the chronic irradiation-induced injury of parotid gland at Grade 1 ,2, 3 was 100%, 0%, 0% with Gln/Gln, 93.9%, 6.1%, 0% with Arg/Gln , 80.9%, 14.3%, 4.7% with Arg/Arg genotype respectively. All above results showed there was no significant difference among the three genotypes (P>0.05).Conclusion:(1) The rate of tumor regression in NPC patients with Gln/Gln genotypes is higher than the other two genotypes at the mid-course and the end of radiotherapy. Its single nucleotide polymorphisms could be a sign of predicting the curative effect at the mid-course and the end of radiotherapy in NPC patients.(2) The the curative effect of radiotherapy in NPC among the three genotypes after 3 months is basically the same, it is mainly related to the clinical stage of nasopharyngeal carcinoma.(3) There is no relativity between the single nucleotide polymorphisms of XRCC1 Codon399 and the acute and chronic irradiation-induced injury in normal tissues.更多还原