CAJ | 학술논문

目的探讨细胞紧密连接基因claudin-4在子宫内膜癌发病和化疗中的作用.方法选择62份子宫内膜样腺癌(EEC)、30份不典型增生子宫内膜和60份正常子宫内膜组织,免疫组化方法和实时荧光定量PCR技术检测claudin-4蛋白、mRNA的表达.90只雌性BALB/c裸鼠腋下注射EEC细胞系Ishikawa,构建人子宫内膜癌裸鼠模型,随机分组进行顺铂(顺铂组)和紫杉醇(紫杉醇组)的腹腔注射化疗(以腹腔注射生理盐水作为对照组),测量化疗前后裸鼠体质量及移植瘤体积的变化,实时荧光定量PCR技术和免疫组化方法检测化疗前后claudin-4 mRNA和蛋白表达的变化.结果在62份EEC中,21份claudin-4蛋白的免疫组化染色呈中等强度,41份呈强阳性表达；30份不典型增生子宫内膜中8份呈弱阳性染色,16份显示中等强度染色,6份显示强阳性染色；60份正常子宫内膜中,28份呈弱阳性染色,32份无claudin-4蛋白的表达.实时荧光定量PCR检测显示,claudin-4 mRNA在EEC、不典型增生内膜和正常子宫内膜中的表达量分别为170±12、89±15和18±3,claudin-4 mRNA在EEC中的表达量高于不典型增生和正常子宫内膜,分别比较,差异均有统计学意义(P＜0.05).子宫内膜癌裸鼠模型检测发现,紫杉醇组和对照组裸鼠化疗前后移植瘤体积及claudin-4的表达均无显著变化；在顺铂组,化疗前移植瘤体积为(0.73±0.12) cm3,化疗后明显缩小至(0.51±0.21)cm3.实时PCR检测显示,顺铂组化疗前、后claudin-4 mRNA的表达量分别为273±27和153±35,claudin-4 mRNA的表达显著降低.结论 claudin-4的过度表达可能与子宫内膜癌的恶性行为有关,claudin-4可作为子宫内膜癌治疗疗效的监测指标.
목적 탐토세포긴밀련접기인claudin-4재자궁내막암발병화화료중적작용.방법 선택62빈자궁내막양선암(EEC)、30빈불전형증생자궁내막화60빈정상자궁내막조직,면역조화방법화실시형광정량PCR기술검측claudin-4단백、mRNA적표체.90지자성BALB/c라서액하주사EEC세포계Ishikawa,구건인자궁내막암라서모형,수궤분조진행순박(순박조)화자삼순(자삼순조)적복강주사화료(이복강주사생리염수작위대조조),측양화료전후라서체질량급이식류체적적변화,실시형광정량PCR기술화면역조화방법검측화료전후claudin-4 mRNA화단백표체적변화.결과 재62빈EEC중,21빈claudin-4단백적면역조화염색정중등강도,41빈정강양성표체；30빈불전형증생자궁내막중8빈정약양성염색,16빈현시중등강도염색,6빈현시강양성염색；60빈정상자궁내막중,28빈정약양성염색,32빈무claudin-4단백적표체.실시형광정량PCR검측현시,claudin-4 mRNA재EEC、불전형증생내막화정상자궁내막중적표체량분별위170±12、89±15화18±3,claudin-4 mRNA재EEC중적표체량고우불전형증생화정상자궁내막,분별비교,차이균유통계학의의(P＜0.05).자궁내막암라서모형검측발현,자삼순조화대조조라서화료전후이식류체적급claudin-4적표체균무현저변화；재순박조,화료전이식류체적위(0.73±0.12) cm3,화료후명현축소지(0.51±0.21)cm3.실시PCR검측현시,순박조화료전、후claudin-4 mRNA적표체량분별위273±27화153±35,claudin-4 mRNA적표체현저강저.결론 claudin-4적과도표체가능여자궁내막암적악성행위유관,claudin-4가작위자궁내막암치료료효적감측지표.
Objective To clarify the role of claudin-4 in endometrial tumorigenesis and explore claudin-4 be as potentially useful agent in the treatment of endometrial carcinoma.Methods The expression of claudin-4 in 62 endonetrioid endometrial carcinoma (EEC),30 atypical hyperplasia endometrial tissue and 60 human normal endometrium was determined using immunohistochemistry and realtime PCR.Ninety female BALB/c mice were transplanted with Ishikawa endometrial cancer cells,which were divided into three groups with different intraperitoneal treatments with cisplatin,paclitaxel and saline solution.After the observation period,the tumors were extracted and stained with monoclonal antibody against claudin-4.The messenger RNA expression of claudin-4 was also detected using real-time PCR.Results Among the EEC samples,34％ (21/62) showed medium staining for claudin-4 and 66％ (41/62) showed intense staining.In atypical hyperplasia group,27％ (8/30) showed weak staining,53％ (16/30) showed medium staining and 20％ (6/30) showed intense staining for claudin-4.Of the normal endometrial tissue,47％ (28/60) showed weak staining and 53％ (32/60) showed no staining for claudin-4.According to real-time PCR,the relative quantity of claudin-4 was 170 ± 12 in EEC group,89 ± 15 in atypical hyperplasia group and 18 ± 3 in normal endometrium.Compared with those in atypical hyperplasia group and normal endometrium group,the protein and mRNA expression of claudin-4 were significantly increased in the group of EEC (all P ＜ 0.05).In the study of Ishikawa xenografts,no significant changes in tumor volume and claudin-4 expression were shown in paclitaxel group compared with that in the control group.Nevertheless,a significant reduction of the tumor growth and a significant decrease in claudin-4 expression were observed in cisplatin group.After cisplatin treatment,the tumor volume was significantly decreased [(0.51 ±0.21) versus (0.73 ±0.12) cm3],and the mRNA expression of claudin-4 was also significantly decreased (153 ± 35 versus 273 ± 27).Conclusion These results demonstrate that claudin-4 is strongly expressed in EEC,which may be a useful biomarker to monitor the effects of chemotherapy in patients with endometrial carcinoma.