Background:The sonic hedgehog(SHH)pathway is an important signaling pathway for neural tube closure.GLI family zinc finger 2(GLI2)is the major activation mediator of the SHH pathway;however,no single-nucleotide polymo...Background:The sonic hedgehog(SHH)pathway is an important signaling pathway for neural tube closure.GLI family zinc finger 2(GLI2)is the major activation mediator of the SHH pathway;however,no single-nucleotide polymorphisms(SNPs)in GLI2 have been reported to be associated with human neural tube defects(NTDs)to date.Here,we evaluated a mutation in GLI2 in the Han Chinese population.Methods:We used SNPscan to genotype rs3738880 in the GLI2 coding region.We then investigated the function of this gene by Western blotting and dual-luciferase assays.Results:In this study,we found that the GLI2 missense variant rs3738880 significantly increased the risk of NTDs in the Han Chinese population via association studies in a cohort of 254 patients and 277 controls from Shanxi Province(odds ratio[OR]=1.89,95%confidence interval[CI]=1.28-2.80,P=0.0012).Additional stratified analyses demonstrated that rs3738880 was significantly related to spina bifida(114 cases,OR=2.01,95%CI=1.19–3.38,P=0.0067).Functional analysis revealed that rs3738880 did not affect GLI2 protein stability and significantly increased SHH activity because of the introduction of a potential phosphorylation site in GLI2.Conclusion:rs3738880 was a risk factor for NTDs in the Han Chinese population.展开更多
文摘目的探讨5-氟尿嘧啶(5-FU)对结肠癌大鼠增殖细胞核抗原(PCNA)表达及端粒酶(hTRT)活性的影响。方法取无特殊病原菌级标准健康雄性裸大鼠36只,10周龄,体重200~230 g。配制结肠癌细胞LoVo单细胞悬液腹腔注射建立结肠癌大鼠模型。将建模成功的30只结肠癌大鼠随机平分为3组-模型组、5-氟尿嘧啶1组、5-氟尿嘧啶2组,模型组予以0.3 mL 0.9%氯化钠溶液灌胃治疗;5-氟尿嘧啶1组、5-氟尿嘧啶2组给予腹腔注射5-氟尿嘧啶,剂量分别为30、60 mg/kg,1次/d,1周3次,共应用4周。测定与记录3组大鼠治疗第2周与第4周的肿瘤体积。观察与记录3组大鼠治疗第4周的结肠癌转移情况,包括局部淋巴结转移、肝转移、肺转移、腹膜转移等。采用免疫组织化学法检测平均单个视野下PCNA阳性细胞、hTRT阳性细胞比率。蛋白质印迹法检测GLI家族锌指蛋白1(Gli1)蛋白、血管内皮生长因子(VEGF)蛋白的相对表达水平。结果5-氟尿嘧啶1组、5-氟尿嘧啶2组治疗第2周与第4周的肿瘤体积均低于模型组(P<0.05),5-氟尿嘧啶2组低于5-氟尿嘧啶1组,差异均有统计学意义(P<0.05)。5-氟尿嘧啶1组、5-氟尿嘧啶2组治疗第4周的结肠癌局部淋巴结转移、肝转移、肺转移、腹膜转移率均低于模型组(P<0.05),5-氟尿嘧啶2组低于5-氟尿嘧啶1组,差异均有统计学意义(P<0.05)。5-氟尿嘧啶1组、5-氟尿嘧啶2组治疗第4周的PCNA、hTRT阳性细胞比率和Gli1、VEGF蛋白相对表达水平均低于模型组(P<0.05),5-氟尿嘧啶2组低于5-氟尿嘧啶1组,差异均有统计学意义(P<0.05)。结论5-氟尿嘧啶在结肠癌大鼠的应用具有很好的抑瘤效果,也可降低周围组织转移率,可抑制Gli1、VEGF蛋白的表达,其作用机制与抑制PCNA表达及hTRT活性具有相关性。
基金This work was supported by grants from the National Key Basic Research Program of China[2016YFC1000502 to H.Wang and X Yang]the National Natural Science Foundation of China[81430005 to H Wang,81472050 to X Yang]Open project of National Population and Family Planning Key Laboratory of Contraceptive drugs and Devices[2016KF04 for X Yang].
文摘Background:The sonic hedgehog(SHH)pathway is an important signaling pathway for neural tube closure.GLI family zinc finger 2(GLI2)is the major activation mediator of the SHH pathway;however,no single-nucleotide polymorphisms(SNPs)in GLI2 have been reported to be associated with human neural tube defects(NTDs)to date.Here,we evaluated a mutation in GLI2 in the Han Chinese population.Methods:We used SNPscan to genotype rs3738880 in the GLI2 coding region.We then investigated the function of this gene by Western blotting and dual-luciferase assays.Results:In this study,we found that the GLI2 missense variant rs3738880 significantly increased the risk of NTDs in the Han Chinese population via association studies in a cohort of 254 patients and 277 controls from Shanxi Province(odds ratio[OR]=1.89,95%confidence interval[CI]=1.28-2.80,P=0.0012).Additional stratified analyses demonstrated that rs3738880 was significantly related to spina bifida(114 cases,OR=2.01,95%CI=1.19–3.38,P=0.0067).Functional analysis revealed that rs3738880 did not affect GLI2 protein stability and significantly increased SHH activity because of the introduction of a potential phosphorylation site in GLI2.Conclusion:rs3738880 was a risk factor for NTDs in the Han Chinese population.
