OCT4在顺铂治疗睾丸胚胎癌中的作用  

The role of OCT4 in cisplatin treated of testicular embryonal carcinoma

作　　者：林杰[1] 谢筱筱[2] 虞相博 秦乐[1] LIN Jie;XIE Xiao-xiao;YU Xiang-bo;QIN Le(Department of Pediatric Surgery,The Second Affiliated Hospital and Yuying Children's Hospital of Wenzhou Medical University,Wenzhou325000,China;Department of Medical Imaging,The Second Affiliated Hospital and Yuying Children's Hospital of Wenzhou Medical University,Wenzhou 325000,China)

机构地区：[1]温州医科大学附属第二医院育英儿童医院小儿外科,温州325000 [2]温州医科大学附属第二医院育英儿童医院放射影像科,温州325000

出　　处：《现代免疫学》2020年第6期446-453,共8页Current Immunology

基　　金：温州市科技局项目(Y20190438)。

摘　　要：为探讨胚胎转录因子八聚体结合转录因子4(octamer-binding transcription factor 4,OCT4)在顺铂治疗睾丸胚胎癌中的作用,采用体外细胞实验评估顺铂对OCT4沉默睾丸胚胎癌细胞株的影响。用500 ng/μL顺铂处理肿瘤细胞后,MTT法检测OCT4沉默/不沉默条件下2012EP和833K-E细胞株的增殖能力,划痕实验检测细胞的迁移能力;采用细胞迁移侵袭实验和结晶紫染色法检测细胞侵袭能力;FACS检测细胞周期分布情况;Western blotting分别检测OCT4沉默/不沉默条件下肿瘤发生相关通路中几种关键蛋白的表达水平。结果显示,与对照组相比,顺铂反增不同浓度OCT4沉默细胞的存活率(P<0.01)。经500 ng/μL顺铂处理后,OCT4沉默细胞的增殖能力是对照组的2~3倍(P<0.001),迁移能力是对照组的2倍(P<0.001),侵袭能力则是约1.5倍(P<0.001)。此外,OCT4沉默上调了参与细胞增殖、迁移、转移和细胞周期调控的重要蛋白的表达水平,而OCT4蛋白表达水平与β连环蛋白呈正相关。由此,OCT4可能成为顺铂治疗睾丸胚胎癌的辅助靶点,其下调或有助于减少化疗中顺铂耐药和随后的疾病复发。To study the role of embryonal transcriptor octamer-binding transcription factor 4(OCT4) in cisplatin treated of testicular embryonal carcinoma, in vitro assays were employed to assess the effect of cisplatin treatment on testicular embryonal carcinoma cell lines under OCT4 silencing condition. Following treatment with 500 ng/μL cisplatin, MTT assay was used to examine cell proliferation of 2012 EP and 833 K-E cells with or without OCT4 silencing, while wound healing assay was used to examine cell migration ability. Transwell assay and crystal violet staining were employed to measure cell invasive capacity, whereas the distribution pattern of cell cycle was assessed by FACS. The expression levels of several critical components in tumorigenicity related pathways with or without OCT4 silencing were determined by Western blotting. Cisplatin enhanced OCT4-silenced cell viability at different concentrations(P<0.01) when compared to control cells. Upon treatment with 500 ng/μL cisplatin, the cell proliferation rate was 2 to 3 folds of the control group(P<0.001), 2 folds of the control group in cell migration capacity(P<0.001), and about 1.5 folds of the control group in invasive capacity(P<0.001). In addition, OCT4 silencing upregulated the expression level of the proteins involved in cell proliferation, migration, metastasis and cell cycle regulation. The results suggest that OCT4 may serve as a therapeutic target for testicular embryonal carcinoma treatment in combination with cisplatin by modulating OCT4 expression level. The study indicates that OCT4 downregulation relieves to cisplatin resistance in chemotherapy and subsequent disease relapse.

关 键 词：八聚体结合转录因子4 顺铂 耐药 睾丸胚胎癌 化疗 

分 类 号：Q955[生物学—动物学]