蛋白S缺乏致肺栓塞患者的PROS1基因突变
发布时间：2018-09-01 16:09
【摘要】：背景:静脉血栓栓塞(VTE)是由血管壁的损伤,血流速度减缓或停止,以及血液的高凝状态引起的。包括下肢深静脉血栓(DVT)和肺栓塞(PE)。近年来,肺栓塞因其发病急,临床表现不典型,死亡率高等特点越发被人们重视。目前遗传性蛋白质S缺乏(PSD)是静脉血栓栓塞的已确立的危险因素。PSD是具有不完全外显率的常染色体显性遗传疾病,与编码蛋白S基因(PROS1)突变或多态性相关,该蛋白S基因共16个外显子,跨越101kb,位于3q11.1。目前国内外已经鉴别出大约200余种PROS1突变引起PS合成或功能的改变。但是,关于蛋白S缺陷症导致肺栓塞的报道并不多见,对符合条件的患者行PROS1基因检测的报道较少。目的:通过对蛋白S缺乏合并肺动脉栓塞的患者行PROS1的基因检测和临床表型诊断,找到突变位点,归纳突变类型。方法:选取吉林大学白求恩第一临床医院自2011年6月至2016年12月期间,行肺动脉CTA检查明确诊断为肺栓塞的患者中,其游离蛋白S活性低于50%的患者40例,同时选取游离蛋白S活性为正常的患者2例。收集他们的外周血10ml,室温下离心分离血细胞及血浆,血浆应用原理凝固方法检测游离蛋白S活性,应用ELISA方法检测游离蛋白S定量,总蛋白S定量;血细胞提取DNA,用于PCR扩增和测序。将患者蛋白S缺乏的情况分为3种类型。I型的特征在于患者血液中总PS、游离PS抗原水平及PS活性同等降低(量的减少);II型的特征在于患者血液中PS的功能活性降低,而总的及游离的PS抗原水平仍正常(质的减少);III型的特征在于总PS抗原水平在正常范围内,而游离PS抗原的水平减少。定位测得的PROS1序列的突变位点,并归纳总结。结果：游离蛋白S活性低于50%的患者40例,其中男性18人(45%),女性22人(55%),男性年龄为(54.88±20.42)岁,女性年龄为(56.59±19.09)岁。其中,遗传性蛋白S缺乏分型:I型患者:22人(55%),II型患者:8人(20%),III型患者:4人(10%),不能分型患者:6人(15%)。行PROS1基因扩增和测序,我们在编码序列上发现了21种不同位点的突变,主要分布在2、5、8、10、12、14、16号外显子上。其中15种突变发生氨基酸的改变,5种突变氨基酸未改变,1种突变成终止密码。在非编码序列上我们同样找到了15种不同位点的突变,主要分布在16号外显子上。蛋白S活性正常的患者2例,其中男性1人,年龄为60岁,女性1名,年龄为59岁。在编码序列上发现了15种不同位点的突变,主要分布在2、5、10号外显子上。其中12种突变发生氨基酸的改变,2种突变氨基酸未改变,1种突变成终止密码。在非编码序列上我们同样找到了10种不同位点的突变,主要分布在16号外显子上。2例PS活性正常的患者检测的结果一致,并且在40例PSD患者中均能找到。40例PSD患者与其进行结果相比:8号外显子第86位C全部发生了c.783 CT(pro261 pro)突变,但是转录的氨基酸仍为脯氨酸(pro)未变。16号外显子第131位A发生了c.2097 AG(pro 699 pro)突变,同样为同义突变,但是这种突变并未在40例患者中都发生(36/40)。有3例患者12号外显子第32位的G发生了c.1283 GA(ser 428 asn)突变,导致氨基酸由丝氨酸(ser)突变成天冬酰胺(asn);1例患者12号外显子第140位G发生c.1391 GA(arg 464 gin)突变,导致氨基酸由精氨酸(arg)突变成谷氨酰胺(gin);1例患者12号外显子第146位T发生c.1397 TC(val 466 ala)突变,导致氨基酸由缬氨酸(val)突变成丙氨酸(ala)。2例患者14号外显子第2位T发生了c.1590 TC(asn 530 asn)突变;但是转录的氨基酸仍为天冬酰胺(asn)未变。结论:1、在40例遗传性蛋白S缺乏导致肺栓塞的患者的编码序列上发现了2种不同位点的同义突变,位于8号外显子第86位C全部发生了c.783 CT(pro 261 pro)突变(40/40),16号外显子第131位A发生了c.2097 AG(pro 699 pro)突变(36/40),这些突变可能引起PSD的发生。2、我们发现了3例患者发生了c.1283 GA(ser 428 asn)突变(3/40),1例患者发生c.1391 GA(arg 464 gin)突变(1/40),1例患者发生c.1397 TC(val466 ala)突变(1/40),2例患者发生了c.1590 TC(asn 530 asn)突变(2/40)。
[Abstract]:BACKGROUND: Venous thromboembolism (VTE) is caused by vascular wall injury, slowing or stopping of blood flow, and hypercoagulable state of blood, including deep venous thrombosis (DVT) and pulmonary embolism (PE). Recently, pulmonary embolism (PE) has attracted more and more attention because of its acute onset, atypical clinical manifestations and high mortality. Poor (PSD) is an established risk factor for venous thromboembolism (VTE). PSD is an autosomal dominant inherited disease with incomplete penetrance and is associated with mutation or polymorphism of the coding protein S gene (PROS1). The protein S gene has 16 exons, spanning 101 kb, and is located in 3q11.1. At present, more than 200 mutations of PROS1 have been identified to cause PS at home and abroad. However, there are few reports about protein S deficiency leading to pulmonary embolism, and few reports about the detection of PROS1 gene in eligible patients. From June 2011 to December 2016, 40 patients with pulmonary embolism diagnosed by CTA were selected from Bethune First Hospital of Jilin University. Their free protein S activity was lower than 50% and 2 patients with normal free protein S activity were selected. Their peripheral blood was collected for 10 ml and centrifuged at room temperature. The free protein S activity was detected by the principle coagulation method, the free protein S and the total protein S were detected by the ELISA method, and the DNA was extracted from the blood cells for PCR amplification and sequencing. Type II was characterized by a decrease in the activity of PS in the blood, while the overall and free PS antigen levels remained normal (qualitative reduction); Type III was characterized by a decrease in total PS antigen levels within the normal range, while free PS antigen levels. Protein S activity was lower than 50% in 40 patients, including 18 males (45%) and 22 females (55%). The male age was (54.88 20.42) and the female age was (56.59 Sequencing, we found 21 different mutations in the coding sequence, mainly distributed in exon 2, 5, 8, 10, 12, 14 and 16. Among them, 15 mutations occurred in amino acids, 5 mutation amino acids remained unchanged, and 1 mutation became a termination code. Exon 2. Two patients with normal protein S activity, one male, 60 years old, one female, 59 years old, were found. 15 mutations were found in the coding sequence, mainly distributed in exons 2, 5 and 10. Among them, 12 mutations occurred in amino acids, 2 mutations in amino acids remained unchanged, and one mutation became a termination code. In the non-coding sequence, we also found 10 different mutations, mainly in exon 16. 2 patients with normal PS activity showed the same results, and 40 patients with PSD were found. 40 patients with PSD compared with the results: all of the 86th C of exon 8 had C. 783 CT (pro261 pro) mutation, but the mutation was reversed. The amino acid in exon 131 A of exon 16 was c.2097 AG (pro 699 pro) mutation, which was also synonymous with the mutation. However, this mutation did not occur in all 40 patients (36/40). In 3 patients, c.1283 GA (ser 428 asn) mutation occurred in exon 12 32 G, resulting in amino acid mutation by ser. Asparagine (asn); c.1391 GA (arg 464 gin) mutation at position 140 of exon 12 in 1 patient, resulting in amino acid mutation from Arg to glutamine (gin); c.1397 TC (val 466 ala) mutation at position 146 of exon 12 in 1 patient, resulting in amino acid mutation from valine (val) to ALA in exon 14 in 2 patients C. 1590 TC (asn 530 asn) mutation occurred in the 2nd T, but the transcriptional amino acid was asparagine (asn) unchanged. Conclusion: 1. Two different mutations were found in the coding sequence of 40 patients with pulmonary embolism caused by inherited protein S deficiency. All of the 86th C in exon 8 had C. 783 CT (pro 261 pro) mutation (40/40). C. 2097 AG (pro 699 pro) mutation (36/40) occurred in 131 A of exon 16. These mutations may cause PSD. 2. We found that 3 patients had C. 1283 GA (ser 428 asn) mutation (3/40), 1 patient had C. 1391 GA (arg 464 gin) mutation (1/40), 1 patient had C. 1397 TC (val466 ala) mutation (1/40), and 2 patients had C. 1397 TC (val466 ala) mutation (1/40). .1590 TC (ASN 530 ASN) mutation (2/40).
【学位授予单位】：吉林大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R563.5
本文编号：2217696
[Abstract]:BACKGROUND: Venous thromboembolism (VTE) is caused by vascular wall injury, slowing or stopping of blood flow, and hypercoagulable state of blood, including deep venous thrombosis (DVT) and pulmonary embolism (PE). Recently, pulmonary embolism (PE) has attracted more and more attention because of its acute onset, atypical clinical manifestations and high mortality. Poor (PSD) is an established risk factor for venous thromboembolism (VTE). PSD is an autosomal dominant inherited disease with incomplete penetrance and is associated with mutation or polymorphism of the coding protein S gene (PROS1). The protein S gene has 16 exons, spanning 101 kb, and is located in 3q11.1. At present, more than 200 mutations of PROS1 have been identified to cause PS at home and abroad. However, there are few reports about protein S deficiency leading to pulmonary embolism, and few reports about the detection of PROS1 gene in eligible patients. From June 2011 to December 2016, 40 patients with pulmonary embolism diagnosed by CTA were selected from Bethune First Hospital of Jilin University. Their free protein S activity was lower than 50% and 2 patients with normal free protein S activity were selected. Their peripheral blood was collected for 10 ml and centrifuged at room temperature. The free protein S activity was detected by the principle coagulation method, the free protein S and the total protein S were detected by the ELISA method, and the DNA was extracted from the blood cells for PCR amplification and sequencing. Type II was characterized by a decrease in the activity of PS in the blood, while the overall and free PS antigen levels remained normal (qualitative reduction); Type III was characterized by a decrease in total PS antigen levels within the normal range, while free PS antigen levels. Protein S activity was lower than 50% in 40 patients, including 18 males (45%) and 22 females (55%). The male age was (54.88 20.42) and the female age was (56.59 Sequencing, we found 21 different mutations in the coding sequence, mainly distributed in exon 2, 5, 8, 10, 12, 14 and 16. Among them, 15 mutations occurred in amino acids, 5 mutation amino acids remained unchanged, and 1 mutation became a termination code. Exon 2. Two patients with normal protein S activity, one male, 60 years old, one female, 59 years old, were found. 15 mutations were found in the coding sequence, mainly distributed in exons 2, 5 and 10. Among them, 12 mutations occurred in amino acids, 2 mutations in amino acids remained unchanged, and one mutation became a termination code. In the non-coding sequence, we also found 10 different mutations, mainly in exon 16. 2 patients with normal PS activity showed the same results, and 40 patients with PSD were found. 40 patients with PSD compared with the results: all of the 86th C of exon 8 had C. 783 CT (pro261 pro) mutation, but the mutation was reversed. The amino acid in exon 131 A of exon 16 was c.2097 AG (pro 699 pro) mutation, which was also synonymous with the mutation. However, this mutation did not occur in all 40 patients (36/40). In 3 patients, c.1283 GA (ser 428 asn) mutation occurred in exon 12 32 G, resulting in amino acid mutation by ser. Asparagine (asn); c.1391 GA (arg 464 gin) mutation at position 140 of exon 12 in 1 patient, resulting in amino acid mutation from Arg to glutamine (gin); c.1397 TC (val 466 ala) mutation at position 146 of exon 12 in 1 patient, resulting in amino acid mutation from valine (val) to ALA in exon 14 in 2 patients C. 1590 TC (asn 530 asn) mutation occurred in the 2nd T, but the transcriptional amino acid was asparagine (asn) unchanged. Conclusion: 1. Two different mutations were found in the coding sequence of 40 patients with pulmonary embolism caused by inherited protein S deficiency. All of the 86th C in exon 8 had C. 783 CT (pro 261 pro) mutation (40/40). C. 2097 AG (pro 699 pro) mutation (36/40) occurred in 131 A of exon 16. These mutations may cause PSD. 2. We found that 3 patients had C. 1283 GA (ser 428 asn) mutation (3/40), 1 patient had C. 1391 GA (arg 464 gin) mutation (1/40), 1 patient had C. 1397 TC (val466 ala) mutation (1/40), and 2 patients had C. 1397 TC (val466 ala) mutation (1/40). .1590 TC (ASN 530 ASN) mutation (2/40).
【学位授予单位】：吉林大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R563.5
【参考文献】
相关期刊论文 前8条
1 侯凤;张涛;李振坤;齐文彦;;血细胞参数在肺栓塞诊断预后中的应用进展[J];临床肺科杂志;2017年01期
2 何献春;寇俊杰;;抗凝药物在急性肺栓塞治疗中的应用进展[J];心脏杂志;2016年03期
3 冯伟;刘晖;;肺栓塞抗凝治疗新进展[J];临床肺科杂志;2015年01期
4 张墨洁;常晓悦;;肺栓塞诊断的研究进展[J];医学综述;2012年04期
5 谷春玲;杨晓红;;蛋白S基因突变及多态性的研究进展[J];现代生物医学进展;2010年10期
6 荆志成;胡大一;;急性肺血栓栓塞症诊断治疗中国专家共识[J];中华内科杂志;2010年01期
7 金佩佩;黄斌伦;戴菁;张浩;余银妹;王学锋;丁秋兰;王鸿利;;两个终止密码子导致的遗传性蛋白C和蛋白S缺陷症[J];诊断学理论与实践;2006年06期
8 臧任丽;孙国超;杨有万;马为民;方志勇;;急性肺动脉栓塞的螺旋CT诊断[J];中国CT和MRI杂志;2006年01期
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