CFTR基因多态性与原发性肝内胆管结石发病风险的关系

梅璇; 吴海聪; 林静; 郑娇龙; 刘邦; 李东良

doi:10.3969/j.issn.1001-5256.2021.12.028

CFTR基因多态性与原发性肝内胆管结石发病风险的关系

DOI: 10.3969/j.issn.1001-5256.2021.12.028

解放军联勤保障部队第九〇〇医院(福建医科大学福总临床医学院) 肝胆内科，福州 350025

基金项目:

福建省社会发展科技引导项目 (2016Y0068);

福建医科大学启航基金 (2017XQ1205)

详细信息

通信作者:
李东良，dongliangli93@163.com

中图分类号: R657.42
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出版历程
- 收稿日期: 2021-05-03
- 录用日期: 2021-05-24
- 出版日期: 2021-12-20

Association of primary intrahepatic lithiasis with the polymorphisms of the cystic fibrosis transmembrane conductance regulator gene

Department of Hepatobiliary Medicine, The 900th Hospital of PLA Joint Logistics Support Force & Fuzong Clinical Medical College of Fujian Medical University, Fuzhou 350025, China

Research funding:

Social Development Science and Technology Project of Fujian Province (2016Y0068);

Fujian Medical University Sailing Fund Program (2017XQ1205)

摘要

摘要: 目的探讨中国汉族人群囊性纤维化跨膜转导调节因子(CFTR)基因常见的多态性位点与原发性肝内胆管结石(PIL)发病的相关性。方法选择2018年6月—2018年11月在解放军联勤保障部队第九〇〇医院就诊的PIL患者104例，以及同期120例健康体检者，采用一代测序法检测CFTR基因的M470V、TG-repeats、Poly-T位点等位基因和基因型，比较2组研究对象的年龄、性别构成比、发病年龄、等位基因及基因型频率，分析CFTR的3个多态性位点与PIL发病风险的相关性。用K-S检验判断连续变量的正态性。呈正态分布计量资料2组间比较用独立样本t检验，计数资料及等位基因及基因型频率比较、Hardy-Weinberg平衡采用χ²检验进行分析。基因型、等位基因与疾病发生风险评估采用二元logistic回归分析。偏离Hardy-Weinberg平衡的位点与PIL发病风险相关性采用调整OR值(OR_adj)表示。结果 PIL组和对照组间的M470V位点上的等位基因(χ²=15.139，P＜0.01)和基因型(χ²=22.8 89，P＜0.01)在2组间差异有统计学意义。TG-repeat、Poly-T两位点上等位基因及个体基因型在2组间差异均无统计学意义(P值均＞0.05)。PIL患者M470V位点上等位基因G频率明显高于对照组(60.10% vs 41.67%，P＜0.01)。与AA基因型个体相比，GG(OR=4.680，P＜0.01)、AG基因型(OR=2.500，P＜0.01)个体发生PIL的风险显著升高。在TG-repeats位点，12TG/13TG基因型个体的PIL发病风险较11TG/12TG个体更高(OR=11.002，P=0.042)；在Poly-T位点7T/5T基因型个体的PIL发病风险较7T/7T个体更低(OR=0.079，P=0.047)。结论中国汉族人群CFTR基因中M470V位点多态性与PIL发病风险独立相关，等位基因G是PIL发病的高危突变。
- 胆结石 /
- 胆管, 肝内 /
- 多态现象, 遗传 /
- 危险因素
Abstract: Objective To investigate the association of common polymorphism loci of the cystic fibrosis transmembrane conductance regulator (CFTR) gene with the onset of primary intrahepatic lithiasis (PIL) in the Chinese Han population. Methods A total of 104 patients with PIL who attended The 900th Hospital of PLA Joint Logistics Support Force from June to November 2018 were enrolled as PIL group, and 120 healthy controls who underwent physical examination during the same period of time were enrolled as control group. Sanger sequencing was used to detect the alleles and genotypes at the M470V, TG-repeats, and Poly-T loci of the CFTR gene. The two groups were compared in terms of age, sex ratio, age of onset, and allele and genotype frequencies, and the association of the above three polymorphism loci of the CFTR gene with the risk of PIL was analyzed. The K-S test was used to determine the normality of continuous variables. The independent samples t-test was used for comparison of normally distributed continuous data between two groups, and the chi-square test was used to compare categorical data and allele/genotype frequencies and analyze Hardy-Weinberg equilibrium. A binary logistic regression analysis was used to investigate the association of genotypes and alleles with the risk of the disease. The association of the loci deviating from Hardy-Weinberg equilibrium with the risk of PIL was expressed as adjusted odds ratio (OR). Results There were significant differences between the PIL group and the control group in the distribution of alleles (χ²=15.139, P < 0.01) and genotypes (χ²=22.889, P < 0.01) at the M470V locus, while there were no significant differences between the two groups in the distribution of alleles and genotypes at the TG-repeats and Poly-T loci (all P > 0.05). The PIL group had a significantly higher frequency of G allele at the M470V locus than the control group (60.1% vs 41.67%, P < 0.01). Compared with the individuals with AA genotype, the individuals with GG and AG genotypes had a significant increase in the risk of PIL (OR=4.680 and 2.500, both P < 0.01). As for the TG-repeats locus, the individuals with 12TG/13TG genotype had a significantly higher risk of PIL than those with 11TG/12TG genotype (OR=11.002, P=0.042), and as for the Poly-T locus, the individuals with 7T/5T genotype had a significantly lower risk of PIL than those with 7T/7T genotype (OR=0.079, P=0.047). Conclusion The M470V polymorphism of the CFTR gene is independently associated with the risk of PIL in the Chinese Han population, and G allele is a high-risk mutation for the onset of PIL.
- Cholelithiasis /
- Bile Ducts, Intrahepatic /
- Polymorphism, Genetic /
- Risk Factors

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表 1 CFTR基因3个位点正反向引物

基因位点	正向引物(5′-3′)	反向引物(5′-3′)
M470V (rs213950)	TTGTGCATAGCAGAGTACCTGAAA	GCTTCTTAAAGCATAGGTCATGTG
Poly-T	CCATGTGCTTTTCAAACTAATTGT	TAAAGTTATTGAATGCTCGCCATG
TG-repeats	CCATGTGCTTTTCAAACTAATTGT	TAAAGTTATTGAATGCTCGCCATG

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表 2 研究对象的一般资料

特征	PIL组 (n=104)	对照组 (n=120)	统计值	P值
年龄(岁)	59.32±14.35	48.47±12.65	t=6.015	＜0.05
男/女(例)	34/70	72/48	χ²=16.667	＜0.05
糖尿病[例(%)]	12(11.53)	18(15.00)	χ²=0.576	0.448
高血压[例(%)]	20(19.23)	13(10.83)	χ²=3.128	0.077
吸烟[例(%)]	28(26.92)	42(35.00)	χ²=1.692	0.193

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表 3 PIL组与对照组M470V位点基因型比较

基因型	PIL组		对照组		OR_adj
基因型	观察值	期望值	观察值	期望值	OR_adj
AA	15	17	52	41	ref
AG	53	50	36	58	2.50
GG	36	37	32	21	4.68
注：OR_adj=(PIL组G1观察值×对照组G2期望值)/(PIL组G2观察值×对照组G1期望值)。G1、G2分别是疾病关联研究中进行对比的两个基因型，G1定义为携带有G等位基因的基因型AG与GG，G2定义为仅携带有A等位基因的基因型AA。

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表 4 PIL组和对照组3个位点等位基因分布

基因位点	等位基因	PIL组(2n=208)	对照组(2n=240)	χ²值	P值¹⁾	OR_adj(95%CI)	P值
M470V	A[例(%)]	83(39.90)	140(58.33)	15.139	＜0.01	ref
	G[例(%)]	125(60.10)	100(41.67)			2.108(1.445~3.077)	＜0.01
TG-repeat	12TG[例(%)]	107(51.44)	134(55.83)	1.778	0.633	ref
	10TG[例(%)]	1(0.48)	3(1.25)			0.417(0.043~4.071)	0.438
	11TG[例(%)]	94(45.19)	97(40.42)			1.214(0.829~1.776)	0.319
	13TG[例(%)]	6(2.89)	6(2.50)			1.252(0.393~3.994)	0.703
Poly-T	7T[例(%)]	206(99.04)	235(97.92)	0.978	0.721	ref
	9T[例(%)]	1(0.48)	2(0.83)			0.570(0.051~6.336)	0.648
	5T[例(%)]	1(0.48)	3(1.25)			0.380(0.039~3.684)	0.404
注：OR_adj指校正性别、年龄后的OR值；1)不同基因位点的等位基因分布的显著性。

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表 5 PIL组和对照组3个位点基因型分布

基因位点	个体基因型	PIL组(n=104)	对照组(n=120)	χ²值	P值¹⁾	OR_adj(95%CI)	P值
M470V	AA[例(%)]	15(14.42)	52(43.33)	22.889	＜0.01	ref
	AG[例(%)]	53(50.96)	36(30.00)			2.500(3.862~24.638)	＜0.01
	GG[例(%)]	36(34.62)	32(26.67)			4.680(2.503~15.879)	＜0.01
TG-repeat	11TG/12TG[例(%)]	52(50.00)	65(54.16)	5.963	0.430	ref
	10TG/11TG[例(%)]	0	3(2.50)				0.999
	10TG/12TG[例(%)]	1(0.96)	2(1.67)			0.980(0.078~12.360)	0.987
	11TG/11TG[例(%)]	20(19.23)	14(11.67)			1.285(0.544~3.032)	0.568
	11TG/13TG[例(%)]	2(1.92)	3(2.50)			2.085(0.172~25.343)	0.564
	12TG/12TG[例(%)]	25(24.04)	31(25.83)			1.635(0.546~4.898)	0.379
	12TG/13TG[例(%)]	4(3.85)	2(1.67)			11.002(1.095~110.565)	0.042
Poly-T	7T/7T[例(%)]	102(98.08)	114(95.00)	1.822	0.779	ref
	7T/9T[例(%)]	1(0.96)	2(1.67)			2.429(0.153~38.567)	0.529
	7T/5T[例(%)]	1(0.96)	3(2.50)			0.079(0.006~0.965)	0.047
	5T/5T[例(%)]	0	1(0.83)
注：OR_adj指校正性别、年龄后的OR值；1)不同基因位点的基因型分布的显著性。

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表 6 PIL组和对照组TG-repeat、Poly-T多态性位点的单倍体基因型分布

单倍体基因型	PIL组(2n=208)	对照组(2n=240)	χ²值	OR_adj(95%CI)	P值
11TG-7T[例(%)]	94(45.19)	98(40.83)	0.865	1.195(0.821~1.739)	0.389
12TG-7T[例(%)]	106(50.96)	127(52.92)	0.171	0.925(0.638~1.341)	0.705
13TG-7T[例(%)]	6(2.88)	5(2.08)	0.299	1.396(0.420~4.643)	0.761
11TG-5T[例(%)]	0	1(0.42)			1.000
12TG-5T[例(%)]	1(0.48)	4(1.67)		0.285(0.032~2.570)	0.379
10TG-9T[例(%)]	1(0.48)	2(0.83)		0.575(0.052~6.386)	1.000
10TG-7T[例(%)]	0	3(1.25)			0.252
注：OR_adj指校正性别，年龄后的OR值。

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