摘要
目的分析细胞毒T淋巴细胞抗原4(CTLA-4)基因-318 T/C、CT60 G/A、+49 G/A多态性与宫颈癌易感性之间的关系。方法以292例宫颈癌患者及355例健康对照者作为研究对象。抽提全血基因组DNA,PCR扩增后用直接测序法检测CTLA-4基因-318 T/C、CT60 G/A、+49 G/A多态性;密度梯度离心法分离外周血单个核细胞(PBMCs),MTT法检测携带不同基因型的PBMCs的增殖能力;ELISA法检测各组血清中IL-2、IL-4、TGF-β的表达水平。结果与CTLA-4-318TT基因型相比,CC基因型降低宫颈癌的发病风险(χ2=7.440,P<0.05);与CTLA-4 CT60GG基因型相比,AA基因型降低宫颈癌的发病风险(χ2=12.165,P<0.05);携带CTLA-4-318 T、CT60 G等位基因的宫颈癌患者增加(χ2分别为13.482和14.138,P均<0.05);而CTLA-4+49 G/A多态性与降低宫颈癌发病风险无关;携带CTLA-4-318TT、TC、CC的PBMCs细胞增殖率差异有统计学意义(F=13.842,P<0.05),而携带CT60、+49不同基因型的PBMCs细胞增殖率差异无统计学意义(F分别为0.578、0.150,P均>0.05)。CTLA-4-318 T/C、CT60 G/A、+49 G/A不同基因型间IL-2、IL-4和TGF-β的水平差异均有统计学意义(P均<0.05)。对宫颈癌组分层分析发现,CTLA-4-318 T/C基因多态性与患者的年龄、绝经与否、HPV感染因素无关(χ2分别为6.310、1.362和4.772,P均>0.05),而与肿瘤分期有关(χ2=18.555,P<0.05)。结论 CTLA-4-318 T/C基因多态性与宫颈癌的患病风险及病情进展有关,其机制可能与调节T细胞的增殖及TH细胞因子的分泌有关。
Objective To investigate the relationship between cytotoxic T-lymphocyte-associated antigen-4（ CTLA-4） genotypes318 T / C,CT60 G / A and ＋ 49 G / A and susceptibility to cervical cancer. Methods Peripheral blood samples from 292 patients with cervical cancer and 355 healthy controls were collected and their genomic DNAs were extracted. The polymorphism of CTLA-4 318 T / C,CT60 G / A and ＋ 49 G / A were detected by direct sequencing after PCR. Meanwhile,the peripheral blood mononuclear cells（ PBMCs）were isolated by the density gradient centrifugation,and their proliferation activity was detected by MTT assay. In addition,serum IL-2,IL-4,and TGF-β levels from cervical cancer patients and healthy controls were detected by ELISA. Results Compared with CTLA-4 genotype 318 TT,CC decreased the onset risk of cervical cancer obviously（ χ^2= 7. 440,P〈0. 05）. Compared with CTLA-4genotype CT60 GG,AA also significantly decreased the onset risk of cervical cancer（ χ^2= 12. 165,P〈0. 05）. The carriers with CTLA-4 318 T and CT60 G alleles suffered from cervical cancer more than those without significantly（ χ^2= 13. 482 and 14. 138,P〈0. 05）. However,the polymorphism of CTLA-4 ＋ 49 G / A was not related to the onset risk of cervical cancer. There was significant difference in the proliferation of PBMCs between the carriers with CTLA-4 318 TT,TC and CC（ F = 13. 842,P〈0. 05）,while there was no any difference in that between the carriers with CTLA-4 CT60 GG,GA and AA,and between those with CTLA-4 ＋ 49 GG,GA and AA（ F = 0. 578 and 0. 150,P〉0. 05）. There were significant difference in serum IL-2,IL-4 and TGF-β levels between different genotypes of CLTA-4 318 T / C,CT60 G / A and ＋ 49 G / A（ P〈0. 05）. In addition,the stratified analysis showed that the polymorphism of CTLA-4 318 T / C was related to the staging of cervical cancer（ χ^2= 18. 555,P〈0. 05）,but unrelated to the patient＇ s age,menopause,and HPV infection（ χ^2= 6. 310,1. 362 and 4. 772,respectively,P〉0. 05）. Conclusion The polymorphism of CTLA-4318 T / C may be related to the onset risk and development of cervical cancer,which might be correlated with the regulation of T cell proliferation and the secretion of TH-type cytokines.
出处
《临床检验杂志》
CAS
CSCD
2015年第2期119-123,共5页
Chinese Journal of Clinical Laboratory Science
基金
四川省医学科学院.四川省人民医院博士基金(30305030569)
