NKG2D基因多态性与喀什地区宫颈癌易感性的关联研究

Association between NKG2D gene polymorphism and cervical cancer susceptibility in Kashgar region

目的研究NKG2D基因多态性与喀什地区宫颈癌易感性的关联。方法选取2020年6月至2021年3月在我院妇科经病理检查明确诊断且均未行任何治疗的各期宫颈癌患者100例作为病例组,选取同期来自本院体检中心及住院患者中排除全身恶性肿瘤的高危型HPV感染的健康妇女100例作为对照组。采集患者组织病理和宫颈分泌物标本。采用Nanodrop测其浓度后进行单核苷酸多态性(single nucleotide polymorphisms,SNP)分型,用聚合酶链反应结合限制性片段长度多态性(PCR-RFLP)技术检测研究对象中NKG2D基因的引物,并测序不同基因型。分析NKG2D基因中的SNP位点rs2617160A>T和rs2255336T>C位点等位基因和基因频率在病例组和对照组患者的分布频率差异,比较NKG2D基因中的SNP位点rs2255336T>C及rs2617160A>T构建的单倍型在病例组和对照组中的分布差异。结果rs2617160A>T和rs2255336T>C位点的基因型在对照组和病例组中分布特征符合Hardy-Weinberg平衡(P>0.05)。多态性位点rs2255336T>C等位基因及基因型在病例组和对照组中分布频率差异不具有统计学意义(χ^(2)=0.287,P=0.592和χ^(2)=0.532,P=0.767)。多态性位点rs2617160A>T等位基因在病例组和对照组中分布频率差异具有统计学意义(χ^(2)=4.511,P=0.034),病例组中等位基因A分布频率较高。多态性位点rs2617160A>T在病例组中AA基因型的分布频率较高(χ^(2)=6.375,P=0.041,OR=0.57,95%CI 0.38~0.90)。病例组和对照组rs2255336T>C和rs2617160A>T之间存在强连锁,D′值分别为0.994和0.991。单倍型rs2255336C-rs2617160A在病例组和对照组中分布频率差异具有统计学意义(P<0.05),且经Bonferroni校正后差异仍具有统计学意义(P<0.05)。单倍型CT和TA在病例组和对照组中的分布频率比较差异不具有统计学意义(P>0.05),且经Bonferroni校正后差异仍不具有统计学意义(P>0.05)。宫颈癌患者的淋巴结转移情况、病理分化程度及临床分期情况在AC、TC、AT单倍型组间比较差异具有统计学意义(P<0.05)。结论NKG2D基因中的SNP位点rs2617160A>T等位基因A及基因型AA可能是宫颈癌发生的风险因素,单倍型CA可能与喀什地区宫颈癌易感性发生有关。

Objective To study the association between NKG2D gene polymorphism and cervical cancer susceptibility in Kashgar.Methods From June 2020 to March 2021,100 patients with cervical cancer diagnosed by pathological examination but no treatment in the department of gynecology were selected as case group,and 100 healthy women with high-risk HPV infection excluding systemic malignant tumors were selected as control group.Samples of pathological tissues and cervical secretion were collected.The concentration of NKG2D gene was measured by Nanodrop,and then single nucleotide polymorphisms(SNPs)were typed.The primers of NKG2D gene were detected by polymerase chain reaction combined with restriction fragment length polymorphism(PCR-RFLP),and different genotypes were sequenced.The difference of allele and gene frequency of SNP sites rs2617160A>T and RS2255336T>C in NKG2D gene was analyzed between case group and control group,and the SNP-based haplotypes at rs2255336T>C and rs2617160A>T in NKG2D gene were compared between case group and control group.Results The distribution of the genotypes of rs2617160A>T and rs2255336T>C in control group and case group was in accordance with Hardy-Weinberg equilibrium(P>0.05).There was no significant difference in the distribution frequency of allele and genotype of rs2255336T>C between case group and control group(χ^(2)=0.287,P=0.592;χ^(2)=0.532,P=0.767).The distribution frequency of rs2617160A>T allele between case group and control group was statistically different(χ^(2)=4.511,P=0.034),and the distribution frequency of allele A in case group was higher.The distribution frequency of AA genotype at rs2617160A>T in case group was higher(χ^(2)=6.375,P=0.041,OR=0.57,95%CI 0.38-0.90).There was a strong linkage between rs2255336T>C and rs2617160A>T,and the values of D′were 0.994 and 0.991 in case group and control group,respectively.The distribution frequency of haplotype rs2255336C-rs2617160A was statistically different between case group and control group(P<0.05),and the difference was still statistically significant after Bonferroni correction(P<0.05).There was no statistical difference in the distribution frequency of haplotype CT and TA between case group and control group(P>0.05),and the difference was still not statistically significant after Bonferroni correction(P>0.05).The lymph node metastasis,the pathological differentiation and the clinical stage of cervical cancer patients were significantly different among AC,TC and AT haplotypes(P<0.05).Conclusion The allele A and genotype AA at SNP rs2617160A>T in NKG2D gene may be risk factors for cervical cancer,and haplotype CA may be related to the susceptibility of cervical cancer in Kashgar.