宫颈癌患者IL-18基因单核苷酸多态性的筛选

目的 筛选宫颈癌患者IL-18基因单核苷酸多态性位点,初步探讨IL-18基因单核苷酸多态性与宫颈癌相关性,为后续选择合适的单核苷酸多态性标志进行宫颈癌风险预测提供参考.方法 常规方法对原发性宫颈癌患者和健康人群46例进行外周血单个核细胞DNA抽提;自行设计引物,对IL-18基因5′端和6个外显子(长约5 kb)进行PCR法,产物采用DNA测序方法进行分析.结果 在IL-18基因5′端2 kp和6个外显子区域内共筛查出7个候选单核苷酸多态性,频率约0.14%,其中rs1946518和rs1946519的单核苷酸多态性基因型在宫颈癌和健康人之间差异有统计学意义(P＜0.05),且二者存在连锁关系;另有3个位点rs360719、rs360717和rs360718也存在连锁关系,可能与宫颈癌呈负相关,但差异无统计学意义(P＞0.05);位于E4的rs11547404和1个未登陆的单核苷酸多态性分别在1例患者中检测到.结论 采用小样本对特定人群IL-18基因进行测序可有效地筛选出候选单核苷酸多态性.初步发现宫颈癌与IL-18基因上游调控区rs1946518和rs1946519单核苷酸多态性有一定关系,为后续利用单核苷酸多态性标志进行宫颈癌风险预测提供了研究基础.

Screening of single nucleotide polymorphisms of IL-18 gene in patients with cervical cancer

Objective To screen single nucleotide polymorphisms(SNPs) of IL-18 gene promoter and exon region in cervical cancer,explore the correlation of IL-18 gene SNPs and cervical cancer,and so as to provide a basis for further screening suitable SNPs labeling forecasting risk for cervical cancer.Methods DNA was Abstracted in peripheral blood mononuclear cell(PBMC) from 20 patients with primary cervical cancer and 46 healthy controls.The primer was designed by ourselves.5'terminal of IL-18 gene and six exons(5 kb approximately) were amplified by PCR.The product was analysed by applying DNA sequencing.Results We found seven candidate SNPs in 5'terminal of IL-18 gene and six exons,in which the differences of rs1946518 and rs1946519 were significant between cervical cancer patients and healthy controls(P<0.05);moreover,the two SNPs were linked with each other.The other three loci(rs360719,rs360717 and rs360718) were linked with each other as well,and appeared to be negatively correlated with cervical cancer.rs11547404 and an unregistered SNPs located in exon 4 were detected in one patient separately.Conclusion Sequencing of IL-18 gene from specific population by adopting small sample is available for screening candidate SNPs.The study reveals initially cervical cancer may be associated with rs1946518 and rs1946519 located in the IL-18 gene upstream regulatory region.It provides a basis for further screening suitable SNPs labeling forecasting risk for cervical cancer.We identified IL-18 gene polymorphisms that may be associated with the risk for cervical cancer.The method of direct sequencing with small samples can screen efficiently the candidate SNPs of IL-18 gene in cervical cancer.