CAJ | 학술논문

目的探讨DNA甲基转移酶1(DNMT1)在子宫颈病变组织和子宫颈癌细胞中的异常表达及其意义.方法选择经病理学确诊的子宫颈鳞状细胞癌(SCC)患者80例、子宫颈上皮内瘤样变(CIN)患者105例(CIN Ⅰ 52例,CINⅡ/Ⅲ53例)和子宫颈正常(NC)者53名,收集其手术或活组织检查子宫颈组织标本.选择子宫颈癌Caski细胞(HPV16阳性)和C33A细胞(HPV阴性)进行体外实验.分别采用Western blot和实时荧光聚合酶链反应(qRT-PCR)法检测子宫颈组织和细胞中DNMT1蛋白和mRNA的表达.结果 DNMT1蛋白和mRNA的表达水平在CIN及SCC组织中均较NC组织高,差异均有统计学意义(F=110.57,P＜0.001；F=2.68,P=0.048)；随着子宫颈病变程度的加重,DNMT1蛋白表达水平呈逐渐上升趋势(x2趋势=50.80,P＜ 0.001),但DNMT1 mRNA的表达在调整子宫颈癌相关因素后,未显示相同趋势(x2趋势=3.63,P＞0.05).Caski细胞DNMT1蛋白和mRNA的表达水平均较C33A细胞高,特别是mRNA的表达水平差异有统计学意义(t=7.134,P=0.002).结论 DNMT1蛋白和mRNA高表达均是导致子宫颈癌和癌前病变发生的危险因素,HPV16感染与DNMT1转录功能异常对子宫颈癌的发生可能有协同效应.
목적 탐토DNA갑기전이매1(DNMT1)재자궁경병변조직화자궁경암세포중적이상표체급기의의.방법 선택경병이학학진적자궁경린상세포암(SCC)환자80례、자궁경상피내류양변(CIN)환자105례(CIN Ⅰ 52례,CINⅡ/Ⅲ53례)화자궁경정상(NC)자53명,수집기수술혹활조직검사자궁경조직표본.선택자궁경암Caski세포(HPV16양성)화C33A세포(HPV음성)진행체외실험.분별채용Western blot화실시형광취합매련반응(qRT-PCR)법검측자궁경조직화세포중DNMT1단백화mRNA적표체.결과 DNMT1단백화mRNA적표체수평재CIN급SCC조직중균교NC조직고,차이균유통계학의의(F=110.57,P＜0.001；F=2.68,P=0.048)；수착자궁경병변정도적가중,DNMT1단백표체수평정축점상승추세(x2추세=50.80,P＜ 0.001),단DNMT1 mRNA적표체재조정자궁경암상관인소후,미현시상동추세(x2추세=3.63,P＞0.05).Caski세포DNMT1단백화mRNA적표체수평균교C33A세포고,특별시mRNA적표체수평차이유통계학의의(t=7.134,P=0.002).결론 DNMT1단백화mRNA고표체균시도치자궁경암화암전병변발생적위험인소,HPV16감염여DNMT1전록공능이상대자궁경암적발생가능유협동효응.
Objective To explore the effects of aberrant expression of DNA methyltransferase 1 (DNMT1) in cervical cancerization tissue and cervical cancer cells.Methods Cervical tissues were collected from 80 cases with a diagnosis of invasive cervix squamous cell carcinoma (SCC),53 cases with high-grade cervical intraepithelial neoplasia (CIN Ⅱ/Ⅲ),52 cases with low-grade cervical intraepithelial neoplasia (CIN Ⅰ)and 53 cases with normal cervix (NC).Meanwhile,Caski (HPV16-positive) and C33A (HPV-negative) cells selected from cervical cancer cell lines were cultured routinely in vitro.The expression of DNMT1 protein and mRNA were examined by Western blot analysis and real-time PCR (qRT-PCR) in the tissues and cells,respectively.Results The levels of DNMT1 protein were 1.33,1.84 and 2.28,and the Ct-ratios (DNMT1/β-actin) of DNMT1 mRNA were 1.27,1.27 and 1.26 in CIN Ⅰ,CIN Ⅱ/Ⅲand SCC group,respectively.Comparing with NC group,the expression of DNMT1 protein or mRNA was elevated in deficient cervical groups,with statistical significance (F =110.57,P ＜ 0.001,F =2.68,P =0.048).The expression levels of DNMT1 protein were increased steadily according to severity of the cervix lesions (x2tend =50.80,P ＜ 0.001),however,the expression of DNMT1 mRNA was not observed the same tendency (x2tend =3.63,P ＞ 0.05).The results from experiment in vitro showed that the levels of DNMT1 protein or mRNA were both higher in Caski cell than in C33A cell,especially for DNMT1 mRNA with significantly difference (t =7.134,P =0.002).Conclusion Aberrant expression of DNMT1 protein or mRNA could link with the risk of cervical cancerization by both transcriptional and posttranscriptional mechanisms.There would be a synergistic effect between overexpression of DNMT1 and HPV16 infection in the progression of cervix carcinogenesis.