脂肪酸合成酶介导骨肉瘤细胞体外“失巢凋亡”抵抗的研究  

A preliminary investigation of FASN-mediated resistance to anokisis in osteosarcoma cells in vitro

作　　者：艾超[1,2] 龙新华[1] 周云飞[1] 刘家明[1] 陈宣银[1] 钟俊桥[3] 刘志礼[1] 

机构地区：[1]南昌大学第一附属医院骨科,南昌330006 [2]广东省江门市新会人民医院骨科,529100 [3]井冈山大学附属医院骨科,江西吉安343000

出　　处：《广东医学》2016年第9期1269-1273,共5页Guangdong Medical Journal

基　　金：国家自然科学基金资助项目(编号:81260400);江西省自然科学基金资助项目(编号:20142BAB205064)

摘　　要：目的比较具有"失巢凋亡"抵抗的骨肉瘤细胞与其亲本细胞中脂肪酸合成酶(FASN)表达和细胞恶型表型的差异。方法 poly-HEMA包被法悬浮培养人骨肉瘤细胞建立人抗失巢凋亡骨肉瘤细胞模型,Western blot、qRT-PCR检测贴壁培养和悬浮培养14 d重贴壁培养的骨肉瘤细胞(U2-OS和143B)中FASN表达水平的差异;Annexin V-FITC/PI双染法流式细胞术,CCK8、Wound healing和Transwell Invasion实验检测细胞凋亡率、细胞增殖、迁徙及侵袭能力的差异。结果常规贴壁培养的U2-OS、143B细胞的增殖、抗凋亡、迁移及侵袭能力显著低于悬浮培养14 d重贴壁培养的U2-OS、143B细胞;FASN蛋白及mRNA在常规贴壁培养的骨肉瘤细胞(U2-OS和143B)中的表达水平显著地低于其在悬浮培养14 d重贴壁培养的细胞中的表达水平。结论 FASN高表达可能通过介导细胞抗"失巢凋亡"促进骨肉瘤细胞的迁徙侵袭。Objective To compare the differences in FASN expression and malignant phenotype between anoikis - resistant human osteosarcoma （OS） cells and their parent cells, and to lay the foundation for further study of the molec- ular mechanisms of promoting OS invasion and metastasis though FASN - mediated anoikis - resistance. Methods Sus- pension culture by coating poly - HEMA on cell culture plates was used to establish the human anoikis - resistant osteosar- coma cell model. The differences in the level of FASN expression between adhere - wall cultured OS cells （ U2 - OS and 143B） and suspension （ 14 d） plus adhere - wall cultured OS cells were measured by Western blot and qRT - PCR assay. The cell apoptotis rate, proliferation, migration and invasion abilities were compared by Annexin V - FITC/PI double stai- ning flow cytometric analysis, CCK8, Wound healing and Transwell invasion assays. Results Compared with the adher- ent OS cells after 14 days＇ suspension culture （suspension ＋ adhere -wall culture）, the adhere -wall cultured OS cells had lower capacity in proliferation, apoptosis -resistance, migration and invasion. The mRNA and protein expressions of FASN were significantly lower in adhere - wall cultured OS cells than in the cells in suspension ＋ adhere - wall culture. Conclusion High expression of FASN may promote invasion and metastasis in OS cells through mediating anoikis - re- sistance.

关 键 词：脂肪酸合酶 失巢凋亡 骨肉瘤 侵袭 转移 

分 类 号：R738.1[医药卫生—肿瘤]