CAJ | 학술논문

目的探讨叶酸缺乏与脆性组氨酸三联体(FHIT)基因异常表达在宫颈癌发生发展中的相互作用.方法选取经病理学确诊的宫颈炎症(CI)患者80例、低度宫颈上皮内瘤样变(CINI)患者55例、高度宫颈上皮内瘤样变(CINⅡ/Ⅲ)患者55例以及宫颈鳞状细胞癌(SCC)患者64例作为研究对象.采用微生物法测定其血清叶酸水平、甲基化特异性PCR检测FHIT基因CpG岛甲基化状况.Western blot法检测宫颈组织中FHIT蛋白的表达水平.同时采用体外细胞试验方法,对宫颈癌细胞CaSki(HPV16阳性)进行叶酸干预,检测不同叶酸浓度下的相关指标的变化.利用SPSS 17.0软件进行相关资料的x2检验、Kruskal-Wallis检验、Spearman秩相关分析,应用相加模型进行交互作用评价.结果随着宫颈病变的加重,血清叶酸含量逐渐降低(H=59.08,P＜0.001),FHIT基因CpG岛甲基化率逐渐升高(趋势检验x2=28.34,P＜0.001),FHIT蛋白表达量逐渐降低(H=50.93,P＜0.001).血清叶酸含量与FHIT蛋白表达量呈正相关(r=0.213,P=0.001),在CIN I、CINⅡ/Ⅲ、SCC组中两者均呈现正相加交互作用.细胞试验显示,随着叶酸浓度增加,宫颈癌细胞的增殖抑制率(r=0.98,P＜0.001)和凋亡率(r=0.99,P＜0.001)逐渐增高,FHIT基因CpG岛甲基化程度逐渐减弱,FHIT蛋白的表达量逐渐升高(r=0.97,P＜0.001).结论叶酸缺乏和FHIT蛋白异常低表达均可增加宫颈癌和癌前病变的发生风险,两者在宫颈癌变中存在正相加交互作用.
목적 탐토협산결핍여취성조안산삼련체(FHIT)기인이상표체재궁경암발생발전중적상호작용.방법 선취경병이학학진적궁경염증(CI)환자80례、저도궁경상피내류양변(CINI)환자55례、고도궁경상피내류양변(CINⅡ/Ⅲ)환자55례이급궁경린상세포암(SCC)환자64례작위연구대상.채용미생물법측정기혈청협산수평、갑기화특이성PCR검측FHIT기인CpG도갑기화상황.Western blot법검측궁경조직중FHIT단백적표체수평.동시채용체외세포시험방법,대궁경암세포CaSki(HPV16양성)진행협산간예,검측불동협산농도하적상관지표적변화.이용SPSS 17.0연건진행상관자료적x2검험、Kruskal-Wallis검험、Spearman질상관분석,응용상가모형진행교호작용평개.결과 수착궁경병변적가중,혈청협산함량축점강저(H=59.08,P＜0.001),FHIT기인CpG도갑기화솔축점승고(추세검험x2=28.34,P＜0.001),FHIT단백표체량축점강저(H=50.93,P＜0.001).혈청협산함량여FHIT단백표체량정정상관(r=0.213,P=0.001),재CIN I、CINⅡ/Ⅲ、SCC조중량자균정현정상가교호작용.세포시험현시,수착협산농도증가,궁경암세포적증식억제솔(r=0.98,P＜0.001)화조망솔(r=0.99,P＜0.001)축점증고,FHIT기인CpG도갑기화정도축점감약,FHIT단백적표체량축점승고(r=0.97,P＜0.001).결론 협산결핍화FHIT단백이상저표체균가증가궁경암화암전병변적발생풍험,량자재궁경암변중존재정상가교호작용.
Objective To explore the interaction between folate deficiency and aberrant expression related to fragile histidine triad (FHIT) gene in the progression of cervical cancerization.Methods A total number of 80 patients with histological diagnosis of cervix inflammation (CI),55 cervical intraepithelial neoplasm Ⅰ (CIN Ⅰ),55 cervical intraepithelial neoplasm Ⅱ/Ⅲ (CIN Ⅱ/Ⅲ) and 64 cervical squamous cell carcinoma (SCC) were included in this study.Levels of serum folate were detected by microbiological assay method and the methylation status of FHIT gene CpG islands was tested by methylation-specific PCR (MSP).FHIT protein levels were measured by Western blot.In vitro,cervical cancer cell lines CaSki (HPV16-positive) was treated with different concentrations of folate.Proliferation and apoptosis of cells,methylation of FHIT gene and the levels of FHIT protein expression were measured in each group.All analyses were performed with SPSS (version 17.0)statistical software.Differences among groups were assessed by chi-square test,Kruskal-Wallis test.Spearman correlation,and the interaction effects were evaluated by additive model.Results The levels of serum folate (H=59.08,P＜0.001) and FHIT protein expression (H=50.93,P＜0.001)decreased gradually along with the severity of cervix lesions,while the methylation rates of FHIT gene CpG islands increased (trend x2=28.34,P＜0.001).Both levels of serum folate levels and FHIT protein expression were positively correlated (r=0.213,P=0.001),with an additive interaction seen between them in CIN Ⅰ,CIN Ⅱ/Ⅲ,SCC groups.In vitro,both rates related to proliferation inhibition (r=0.98,P＜0.001) and apoptosis (r=0.99,P＜0.001) together with the levels of FHIT protein expression (r=0.97,P＜0.001) were all increased gradually with the increase of folate concentration while the methylation status of FHIT gene CpG islands all changed from positive to negative gradually.Conclusion Results from our study revealed that both folate deficiency and FHIT protein aberrant low expression might increase the risk of developing cervical cancer and cervix precancerous lesions,and thus play a synergistic action in the progression of cervical cancerization.