CDKN2B‑AS1基因多态性与克罗恩病发病风险的相关性分析

The associations of cyclin‑dependent kinase inhibitor 2B antisense RNA 1 gene polymorphisms with the risk of Crohn′s disease in Chinese patients

目的探讨细胞周期依赖性激酶抑制因子2B反义链1(CDKN2B‑AS1)基因多态性与克罗恩病(CD)发病风险的相关性。方法选择2012年1月至2021年1月于温州医科大学附属第二医院消化内科收集的207例CD患者[男133例,女74例,年龄(31.6±10.8)岁]及年龄、性别相匹配的545名健康对照者[男337名,女208名,年龄(31.7±9.3)岁]。采用基质辅助激光解吸电离‑时间飞行质谱技术检测两组受试者CDKN2B‑AS1(rs1063192、rs10757274、rs10757278、rs1333048、rs2383207)的基因型。在CD组和健康对照组之间,采用非条件logistic回归分析CDKN2B‑AS1基因多态性的分布差异及其对CD患者临床病理特征的影响。采用Haploview 4.2软件进行连锁不平衡(LD)及单倍型分析。结果CD组中rs1063192的变异基因型(AG+GG)和变异等位基因(G)的频率均高于健康对照组(32.4%比24.8%,P=0.036;18.8%比13.6%,P=0.011)。CD组中rs10757274的纯合子变异基因型(GG)和变异等位基因(G)的频率亦显著高于健康对照组(19.8%比12.8%,P=0.017;45.2%比38.1%,P=0.012)。根据蒙特利尔CD表型分类法对患者进行分层分析发现,(狭窄型+穿透型)CD患者中rs10757278的纯合子变异基因型(GG)和变异等位基因(G)的频率均显著低于非狭窄非穿透型CD患者(13.7%比29.9%,P=0.015;37.7%比50.4%,P=0.022)。上述结果经Bonferroni校正后差异均无统计学意义(均P>0.05)。LD分析结果显示CDKN2B‑AS1基因上rs10757274、rs2383207、rs10757278和rs1333048彼此紧密连锁。CD组中单倍型AGAC的频率显著低于健康对照组(1.5%比4.5%,χ^(2)=7.61,P=0.006),但单倍型GGAC的频率显著高于健康对照组(3.0%比0.6%,χ^(2)=14.25,P<0.001)。进一步分层分析发现,(狭窄型+穿透型)CD患者中单倍型AGAC的频率高于非狭窄非穿透型CD患者(3.1%比0.4%,χ^(2)=5.31,P=0.021)。结论CDKN2B‑AS1基因上rs1063192、rs10757274、rs10757278、rs1333048和rs2383207发生变异可能不会独立影响CD发病风险。rs10757274、rs2383207、rs10757278和rs1333048彼此紧密连锁,虽然携带单倍型AGAC可能降低CD发病风险,但可能提高CD患者发生狭窄或穿透的风险。另外,携带单倍型GGAC可能增加CD发病风险。

Objectives The present study aimed to investigate the associations of cyclin‑dependent kinase inhibitor 2B antisense RNA 1(CDKN2B‑AS1)gene polymorphisms with the risk of Crohn′s disease(CD)in Chinese patients.Methods From January 2012 to January 2021,a total of 207 CD patients and 545 age‑and gender‑matched healthy controls were collected from the Department of Gastroenterology,the Second Affiliated Hospital of Wenzhou Medical University.The genotypes of CDKN2B‑AS1(rs1063192,rs10757274,rs10757278,rs1333048,rs2383207)were determined by matrix‑assisted laser desorption ionization time‑of‑flight mass spectrometry technique.Unconditional logistic regression analysis was used to analyze the differences of CDKN2B‑AS1 polymorphisms between CD patients and healthy controls,as well as their influences on the clinicopathologic characteristics of CD patients.The analyses for linkage disequilibrium and haplotype were further performed by Haploview 4.2 software.Results The variant genotype(AG+GG)and variant allele(G)of rs1063192 were more prevalent in CD patients than in healthy controls(32.4%vs 24.8%,P=0.036;18.8%vs 13.6%,P=0.011).The same conclusions were also drawn for homozygous variant genotype(GG)and variant allele(G)of rs10757274 when CD patients were compared with healthy controls(19.8%vs 12.8%,P=0.017;45.2%vs 38.1%,P=0.012).According to the Montreal Classification Standards,CD patients were stratified into different subgroups.The homozygous variant genotype(GG)and variant allele(G)of rs10757278 were less frequent in the patients with stricturing CD or penetrating CD than in those with non‑stricturing and non‑penetrating CD(13.7%vs 29.9%,P=0.015;37.7%vs 50.4%,P=0.022).However,all the correlations above were no longer significant after Bonferroni′s correction(all P>0.05).The polymorphic loci of rs10757274,rs2383207,rs10757278,and rs1333048 were in close linkage disequilibrium with each other in CDKN2B‑AS1 gene.Compared with healthy controls,the frequency of haplotype AGAC was decreased in CD patients(1.5%vs 4.5%,χ^(2)=7.61,P=0.006),whereas the frequency of haplotype GGAC was obviously increased in CD patients(3.0%vs 0.6%,χ^(2)=14.25,P<0.001).The stratified analysis further showed that the frequency of haplotype AGAC was higher in the patients with stricturing CD or penetrating CD than in those with non‑stricturing and non‑penetrating CD(3.1%vs 0.4%,χ^(2)=5.31,P=0.021).Conclusions The variations of CDKN2B‑AS1(rs1063192,rs10757274,rs10757278,rs1333048,rs2383207)may not independently affect the risk of CD.Among the haplotypes constructed by rs10757274,rs2383207,rs10757278,and rs1333048,the haplotype AGAC may reduce the risk of CD,whereas it may increase the risk of stricturing or penetrating in CD patients.In addition,the haplotype GGAC may increase the risk of CD.