中华预防医学杂志
中華預防醫學雜誌
중화예방의학잡지
CHINESE JOURNAL OF
2012年
12期
1088-1094
,共7页
王金桃%丁玲%郝俊霞%赵维敏%周芩%郝敏
王金桃%丁玲%郝俊霞%趙維敏%週芩%郝敏
왕금도%정령%학준하%조유민%주금%학민
叶酸%宫颈鳞状细胞癌%宫颈上皮内瘤样病变%DNA甲基转移酶1%宫颈癌细胞
葉痠%宮頸鱗狀細胞癌%宮頸上皮內瘤樣病變%DNA甲基轉移酶1%宮頸癌細胞
협산%궁경린상세포암%궁경상피내류양병변%DNA갑기전이매1%궁경암세포
Folic acid%Cervix squamous cell carcinoma%Cervical intraepithelial neoplasia%DNA methyltransferase 1%Cervical cancer cell
目的 探讨叶酸缺乏和DNA甲基转移酶1(DNMT1)表达异常在宫颈癌发生发展中的相互作用.方法 选择2009年9月至2010年5月,在山西某医院确诊的宫颈鳞状细胞癌(SCC)患者80例、宫颈上皮内瘤样变(CIN)患者105例(其中CIN Ⅰ组52例,CIN Ⅱ/Ⅲ组53例)和宫颈炎(CI)患者53例为研究对象,采用放射免疫法(RIA)检测其血清叶酸水平.同时,采用体外实验研究方法,对宫颈癌细胞Caski[人乳头瘤病毒(HPV)16阳性]和C33A(HPV阴性)进行不同浓度叶酸干预.检测宫颈组织和细胞中DNMT1蛋白的表达量和mRNA水平.利用SPSS 16.0软件进行相关资料的t检验、方差分析、x2检验、Spearman相关分析,应用相加效应模型进行交互作用评价.结果 CI组、CIN Ⅰ组、CINⅡ/Ⅲ组和SCC组血清叶酸含量(中位数±四分位数间距)分别为(3.21±1.74)、(3.17±1.91)、(2.83±2.23)、(2.66±1.82) ng/ml(P＜0.05),DNMT1蛋白表达水平((x)±s)分别为0.92±0.29、1.33±0.38、1.84±0.37、2.28±0.55,DNMT1 mRNA表达水平((x)±s)分别为1.33±0.11、1.27±0.12、1.27±0.12、1.26±0.13；随着宫颈病变的加重,叶酸含量逐渐降低,DNMT1蛋白和mRNA表达水平逐渐增高(P值均＜0.05)；低叶酸水平和DNMT1蛋白高表达在宫颈癌及癌前病变组均存在正相加交互作用,交互作用相对超额危险度(RERI)、交互作用归因危险比(API)和交互作用指数(S)均显示正相加效应(CIN Ⅰ组:0.27、0.14、1.40；CIN Ⅱ/Ⅲ组:0.47、0.19、1.46；SCC组:1.60、0.31、1.61).体外研究显示,Caski和C33A细胞的生长抑制率从叶酸干预水平为10 μg/ml时的11.4％和13.6％分别上升至1000 μg/ml时的64.8％和49.4％,随着叶酸水平的升高呈上升趋势(r值分别为0.954、0.969,P值均＜0.05)；DNMT1蛋白的表达量随着叶酸水平的升高而降低(r值分别为-0.859、-0.914,P值均＜0.05),分别从叶酸干预水平10 μg/ml时的1.96和1.92降低至1000 μg/ml时的1.60和1.38;叶酸浓度为1000 μg/ml时,Caski细胞DNMT1蛋白和mRNA的表达水平均较C33A细胞高(t值分别为-4.22、3 50,P值均＜0.05).结论 血清叶酸水平低及DNMT1蛋白和mRNA高表达均可增加宫颈癌和癌前病变的发病风险,叶酸缺乏与DNMT1蛋白高表达有正交互作用；补充叶酸可有效抑制宫颈癌细胞的增殖,逆转DNMT1的异常转录和转录后表达异常.
目的 探討葉痠缺乏和DNA甲基轉移酶1(DNMT1)錶達異常在宮頸癌髮生髮展中的相互作用.方法 選擇2009年9月至2010年5月,在山西某醫院確診的宮頸鱗狀細胞癌(SCC)患者80例、宮頸上皮內瘤樣變(CIN)患者105例(其中CIN Ⅰ組52例,CIN Ⅱ/Ⅲ組53例)和宮頸炎(CI)患者53例為研究對象,採用放射免疫法(RIA)檢測其血清葉痠水平.同時,採用體外實驗研究方法,對宮頸癌細胞Caski[人乳頭瘤病毒(HPV)16暘性]和C33A(HPV陰性)進行不同濃度葉痠榦預.檢測宮頸組織和細胞中DNMT1蛋白的錶達量和mRNA水平.利用SPSS 16.0軟件進行相關資料的t檢驗、方差分析、x2檢驗、Spearman相關分析,應用相加效應模型進行交互作用評價.結果 CI組、CIN Ⅰ組、CINⅡ/Ⅲ組和SCC組血清葉痠含量(中位數±四分位數間距)分彆為(3.21±1.74)、(3.17±1.91)、(2.83±2.23)、(2.66±1.82) ng/ml(P＜0.05),DNMT1蛋白錶達水平((x)±s)分彆為0.92±0.29、1.33±0.38、1.84±0.37、2.28±0.55,DNMT1 mRNA錶達水平((x)±s)分彆為1.33±0.11、1.27±0.12、1.27±0.12、1.26±0.13；隨著宮頸病變的加重,葉痠含量逐漸降低,DNMT1蛋白和mRNA錶達水平逐漸增高(P值均＜0.05)；低葉痠水平和DNMT1蛋白高錶達在宮頸癌及癌前病變組均存在正相加交互作用,交互作用相對超額危險度(RERI)、交互作用歸因危險比(API)和交互作用指數(S)均顯示正相加效應(CIN Ⅰ組:0.27、0.14、1.40；CIN Ⅱ/Ⅲ組:0.47、0.19、1.46；SCC組:1.60、0.31、1.61).體外研究顯示,Caski和C33A細胞的生長抑製率從葉痠榦預水平為10 μg/ml時的11.4％和13.6％分彆上升至1000 μg/ml時的64.8％和49.4％,隨著葉痠水平的升高呈上升趨勢(r值分彆為0.954、0.969,P值均＜0.05)；DNMT1蛋白的錶達量隨著葉痠水平的升高而降低(r值分彆為-0.859、-0.914,P值均＜0.05),分彆從葉痠榦預水平10 μg/ml時的1.96和1.92降低至1000 μg/ml時的1.60和1.38;葉痠濃度為1000 μg/ml時,Caski細胞DNMT1蛋白和mRNA的錶達水平均較C33A細胞高(t值分彆為-4.22、3 50,P值均＜0.05).結論 血清葉痠水平低及DNMT1蛋白和mRNA高錶達均可增加宮頸癌和癌前病變的髮病風險,葉痠缺乏與DNMT1蛋白高錶達有正交互作用；補充葉痠可有效抑製宮頸癌細胞的增殖,逆轉DNMT1的異常轉錄和轉錄後錶達異常.
목적 탐토협산결핍화DNA갑기전이매1(DNMT1)표체이상재궁경암발생발전중적상호작용.방법 선택2009년9월지2010년5월,재산서모의원학진적궁경린상세포암(SCC)환자80례、궁경상피내류양변(CIN)환자105례(기중CIN Ⅰ조52례,CIN Ⅱ/Ⅲ조53례)화궁경염(CI)환자53례위연구대상,채용방사면역법(RIA)검측기혈청협산수평.동시,채용체외실험연구방법,대궁경암세포Caski[인유두류병독(HPV)16양성]화C33A(HPV음성)진행불동농도협산간예.검측궁경조직화세포중DNMT1단백적표체량화mRNA수평.이용SPSS 16.0연건진행상관자료적t검험、방차분석、x2검험、Spearman상관분석,응용상가효응모형진행교호작용평개.결과 CI조、CIN Ⅰ조、CINⅡ/Ⅲ조화SCC조혈청협산함량(중위수±사분위수간거)분별위(3.21±1.74)、(3.17±1.91)、(2.83±2.23)、(2.66±1.82) ng/ml(P＜0.05),DNMT1단백표체수평((x)±s)분별위0.92±0.29、1.33±0.38、1.84±0.37、2.28±0.55,DNMT1 mRNA표체수평((x)±s)분별위1.33±0.11、1.27±0.12、1.27±0.12、1.26±0.13；수착궁경병변적가중,협산함량축점강저,DNMT1단백화mRNA표체수평축점증고(P치균＜0.05)；저협산수평화DNMT1단백고표체재궁경암급암전병변조균존재정상가교호작용,교호작용상대초액위험도(RERI)、교호작용귀인위험비(API)화교호작용지수(S)균현시정상가효응(CIN Ⅰ조:0.27、0.14、1.40；CIN Ⅱ/Ⅲ조:0.47、0.19、1.46；SCC조:1.60、0.31、1.61).체외연구현시,Caski화C33A세포적생장억제솔종협산간예수평위10 μg/ml시적11.4％화13.6％분별상승지1000 μg/ml시적64.8％화49.4％,수착협산수평적승고정상승추세(r치분별위0.954、0.969,P치균＜0.05)；DNMT1단백적표체량수착협산수평적승고이강저(r치분별위-0.859、-0.914,P치균＜0.05),분별종협산간예수평10 μg/ml시적1.96화1.92강저지1000 μg/ml시적1.60화1.38;협산농도위1000 μg/ml시,Caski세포DNMT1단백화mRNA적표체수평균교C33A세포고(t치분별위-4.22、3 50,P치균＜0.05).결론 혈청협산수평저급DNMT1단백화mRNA고표체균가증가궁경암화암전병변적발병풍험,협산결핍여DNMT1단백고표체유정교호작용；보충협산가유효억제궁경암세포적증식,역전DNMT1적이상전록화전록후표체이상.
Objective To explore the interaction of folate deficiency and aberration of DNA methyltransferase 1 (DNMT1) in the progression of cervix carcinogenesis.Methods All clinical samples were collected from 80 patients with cervix squamous cell carcinoma (SCC),105 patients with cervical intraepithelial neoplasm (CIN Ⅰ,n =52 ; CIN Ⅱ / Ⅲ,n =53) and 53 patients with cervix inflammation (CI).The participants were diagnosed by histology at Shanxi Province Tumor Hospital and Second Hospital of Shanxi Mcdical University during the period of September 2009 to May 2010.Meanwhile,cervical cancer cell lines Caski and C33 A were treated with different concentration of folate.Radioimmunoassay (RIA),Western blotting and real-time PCR were used to detect the levels of serum folate,the expression of DNMT1 protein and mRNA,respectively.The data were analyzed by Student t test,ANOVA,chi-square test and Spearman correlation using SPSS statistical package.The correlation strength between factors and cervical canceriation was calculated by OR and 95％ CI value.Interaction effect was evaluated by the application of additive effect model.Results The levels of serum folate (median inter-quartile range) were (2.66 ± 1.82),(2.83 ±2.23),(3.17±1.91) and (3.21±1.74)ng/ml,the levels of DNMT1 protein ((x)±s) were 2.28 ±0.55,1.84 ± 0.37,1.33 ± 0.38 and 0.92 ± 0.29,the Ct-ratio (Ct value of DNMT1/Ct value of β-actin) of DNMT1 mRNA ((x)± s) were 1.26 ± 0.13,1.27 ± 0.12,1.27 ± 0.12 and 1.33 ± 0.11 in the group of SCC,CIN Ⅱ/Ⅲ,CIN Ⅰ and CI,respectively.The results showed that the serum folate levels were descended,and the expression levels of DNMT1 protein (x2tend =50.80,P ＜ 0.05) and mRNA (x2tend =17.63,P ＜ 0.05)were increased steadily with the severity of the cervix lesions.Moreover,our results revealed that there was an additive interaction between folate deficiency and high-expression of DNMT1 protein related to the risk of C1N and SCC.And it showed that the relative excess risk of interaction(RERI),attributable proportion of interaction(API) and synergy index (S) was 0.27,0.14 and 1.40 in CIN Ⅰ group,0.47,0.19,1.46 in CIN Ⅱ/Ⅲ group,1.60,0.31,1.61 in SCC group,respectively.It was found that folate was able to reduce the proliferation of Caski and C33A cells (r values were 0.954 and 0.969,all P values ＜ 0.05),with 11.4％ and 13.6％ of growth inhibition at the concentration of 10 μg/ml,64.8％ and 49.4％ at 1000 μg/ml in Caski and C33A cells,respectively.The result showed there was an inverse correlation between the levels of folate and DNMT1 protein(r values were-0.859 and-0.914,all P values ＜ 0.05),with 1.96 and 1.92 of expression levels at the concentration of 10 μg/ml,and 1.60 and 1.38 at 1000 μg/ml in Caski and C33A cells,respectively.At folate concentration of 1000 μg/ml,the expression of DNMT1 protein or mRNA was higher in Caski cell than in C33A cell (t values were-4.22 and 3.50,all P values ＜0.05).Conclusion Our finding indicated that the low levels of serum folate and high-expression of DNMT1 protein or mRNA seemed to be associated with high risk of cervical cancer and cervix precancerous lesion.Sufficient folate is able to effectively inhibit the growth of cervical cancer cells in vitro,and would counteract transcriptional and posttranscriptional aberration of DNMT1.It suggested that there might be a synergistic action between folate deficiency and aberration of DNMT1 in the progression of cervix carcinogenesis.
