罕见同卵双胞胎侵袭性牙周炎患者易感基因研究

doi:10.12016/j.issn.2096-1456.2020.11.003

摘要/Abstract

摘要： 目的探究易感基因对侵袭性牙周炎发生发展的影响,为进一步探索遗传因素在侵袭性牙周炎中的发病机制提供依据。方法收集并分析两例罹患广泛型侵袭性牙周炎的同卵双胞胎患者的病史及牙周基础治疗前后相关临床指标。聚合酶链式反应-限制性内切酶多态性分析法检测二者基因组中侵袭性牙周炎目标候选易感基因,全外显子测序法分析二者外显子基因成分。结果该同卵双胞胎广泛型侵袭性牙周炎患病程度不同（P < 0.05）,Florida 探诊结果显示妹妹探诊深度（probing depth, PD）、临床附着丧失（clinical attachment loss,CAL）指标高于姐姐,但姐姐探诊出血指数（bleeding on probing,BOP）高于妹妹;且对牙周基础治疗的反应效果不同（P < 0.05）,治疗后,姐姐CAL的改善较明显,BOP下降幅度也更明显,且治疗后姐姐不同袋深的比例均降低,而妹妹降低较少。基因检测结果发现二者均携带具有相同基因型的目标易感基因（IL-1β-511、IL-1β+3953、TNF-α-308、FcγR-IIIb、VDR、ER）;外显子分析结果显示二者基因组存在差异,其中ZFPM1、PTH2、ZFYVE16、LY6G6C 4个可疑的差异性基因可能与二者广泛型侵袭性牙周炎的疾病表型差异相关。结论该同卵双胞胎广泛型侵袭性牙周炎的疾病表型存在差异,二者携带的易感基因增加了其患病的风险,二者突变的差异基因影响了其疾病的表型。

关键词: 广泛型侵袭性牙周炎, 同卵双胞胎, 易感基因, 全外显子测序, 牙周治疗, 基因突变, 拷贝数变异

Abstract:

Abstract Objective Exploring the effects of susceptibility genes on aggressive periodontitis during its occurrence and development lays a foundation for further research on its genetic pathogenesis. Methods Medical history and clinical indicators were collected from monozygotic twins with generalized aggressive periodontitis. Susceptibility genes were detected by polymerase chain reaction-restriction fragment length polymorphisms, and the exon gene components were analyzed by whole-exome sequencing. Results The severity of generalized aggressive periodontitis in the twins was different (P < 0.05). Florida diagnosis results showed that the younger sister’s explored clinical depth (probing depth, PD) and clinical attachment loss (CAL) were higher than those for the elder sister, but her clinical bleeding index (bleeding on probing, BOP) was lower than that of the elder sister. In addition, their responses to periodontal basic treatment were different (P < 0.05). After treatment, the improvement of CAL and decline in BOP were more obvious in the elder sister, and the proportion of different periodontal pocket depths in the elder sister decreased after treatment, while the decrease was slighter in the younger sister. Genetic tests revealed that the twins had the same genotypes at target susceptibility genes (IL-1β-511, IL-1β+3953, TNF-α-308, FcγR-IIIb, VDR, and ER) and differential exon gene components, and that the suspected differential genes, e.g. ZFPM1, PTH2, ZFYVE16, and LY6G6C, might be related to their phenotypic differences. Conclusion These monozygotic twins had different phenotypes of generalized aggressive periodontitis. Their shared susceptibility genes increased the risk of disease, and their different genetic mutations affected the phenotype of the disease.

Key words: aggressive periodontitis, monozygotic twin, susceptibility gene, whole-exome sequencing, periodontal therapy, genetic mutations, copy number variation

中图分类号:

R781.4

刘丽,蓝露芳,郭淑娟,吴亚菲. 罕见同卵双胞胎侵袭性牙周炎患者易感基因研究[J]. 口腔疾病防治, 2020, 28(11): 698-704.

LIU Li,LAN Lufang,GUO Shujuan,WU Yafei. A study of susceptibility genes in rare monozygotic twins with generalized aggressive periodontitis[J]. Journal of Prevention and Treatment for Stomatological Diseases, 2020, 28(11): 698-704.

图/表 7

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Genes	Primer sequences
IL-1β-511	F：5'-CCAAGAGATAGAGTCTCAGATGG-3'
IL-1β-511	R：5'-GGACAAAGTGGAAGACACACAG-3'
IL-1β+3953	F：5'-CTCAGGTGTCTCCAAGAAATCAAA-3'
IL-1β+3953	R：5'-GCTTTTTTGCTGTGAGTCCCG-3'
TNF-α-308	F：5'-GGCAATGGTTTGAGGGC-3'
TNF-α-308	R：5'-GAGGGCGGGAAAGAATCAT-3'
FcγR IIIb（NA1）	F：5'-CAGTGGTTTCACAATGTGAA-3'
FcγR IIIb（NA1）	R：5'-CATGGACTTCTAGCACCG-3'
FcγR IIIb（NA2）	F：5'-CAATGGTACAGCGTGCTT-3'
FcγR IIIb（NA2）	R：5'-CTGTACTCTCCACTGTCGTT-3'
VDR	F：5'-CAGAGCATGGACAGGGAGCAA-3'
VDR	R：5'-GCAACTCCTCATGGCTGAGGTCTC-3'
ER	F：5'-CTGCCACCCTATCTGCTTTTCCTATTCTCC-3'
ER	R：5'-TCTTTCTCTGCCACCCTGGCGTGGATTATCTGA-3'

Genes	Primer sequences
IL-1β-511	F：5'-CCAAGAGATAGAGTCTCAGATGG-3'
IL-1β-511	R：5'-GGACAAAGTGGAAGACACACAG-3'
IL-1β+3953	F：5'-CTCAGGTGTCTCCAAGAAATCAAA-3'
IL-1β+3953	R：5'-GCTTTTTTGCTGTGAGTCCCG-3'
TNF-α-308	F：5'-GGCAATGGTTTGAGGGC-3'
TNF-α-308	R：5'-GAGGGCGGGAAAGAATCAT-3'
FcγR IIIb（NA1）	F：5'-CAGTGGTTTCACAATGTGAA-3'
FcγR IIIb（NA1）	R：5'-CATGGACTTCTAGCACCG-3'
FcγR IIIb（NA2）	F：5'-CAATGGTACAGCGTGCTT-3'
FcγR IIIb（NA2）	R：5'-CTGTACTCTCCACTGTCGTT-3'
VDR	F：5'-CAGAGCATGGACAGGGAGCAA-3'
VDR	R：5'-GCAACTCCTCATGGCTGAGGTCTC-3'
ER	F：5'-CTGCCACCCTATCTGCTTTTCCTATTCTCC-3'
ER	R：5'-TCTTTCTCTGCCACCCTGGCGTGGATTATCTGA-3'

Groups	PD（mm）				CAL（mm）				BOP
Groups	Elder	Younger	t	P	Elder	Younger	t	P	Elder	Younger
Pre-therapy	3.51 ± 1.17	4.65 ± 2.20	5.89	<0.001	3.68 ± 1.29	6.73 ± 2.86	12.57	<0.001	52%	22%
Post-treatment	2.89 ± 0.93	3.79 ± 1.69			2.94 ± 1.00	6.66 ± 2.57			15%	10%
Value of effect	0.63 ± 0.92	0.86 ± 1.56	1.75	0.081	0.75 ± 0.87	0.07 ± 1.45	1.97	0.049
t	5.38	4.02			5.97	0.22
P	<0.001	<0.001			<0.001	0.826

Groups	PD（mm）				CAL（mm）				BOP
Groups	Elder	Younger	t	P	Elder	Younger	t	P	Elder	Younger
Pre-therapy	3.51 ± 1.17	4.65 ± 2.20	5.89	<0.001	3.68 ± 1.29	6.73 ± 2.86	12.57	<0.001	52%	22%
Post-treatment	2.89 ± 0.93	3.79 ± 1.69			2.94 ± 1.00	6.66 ± 2.57			15%	10%
Value of effect	0.63 ± 0.92	0.86 ± 1.56	1.75	0.081	0.75 ± 0.87	0.07 ± 1.45	1.97	0.049
t	5.38	4.02			5.97	0.22
P	<0.001	<0.001			<0.001	0.826

Groups		PD ≤ 4 mm	4 mm<PD<6 mm	PD ≥ 6 mm
Elder	Pre-therapy	44.6%	38.1%	6.5%
	Post-treatment	14.3%	13.7%	0.6%
	Reduced proportion	30.3%	24.4%	5.9%
Younger	Pre-therapy	64.3%	25.0%	39.3%
	Post-treatment	52.3%	35.1%	17.3%
	Reduced proportion	12%	-15.1%	22%