摘要
目的探讨脂肪间充质干细胞是否通过调节EZH2的表达促进卵巢癌细胞生长、侵袭及迁移。方法体外分离、培养健康人大网膜脂肪间充质干细胞并制备脂肪间充质干细胞条件培养基(ADSCs-CM),建立EZH2shRNA稳定转染的卵巢癌细胞系。条件培养基处理稳定转染及未转染的卵巢癌细胞,实时定量PCR及Western blot免疫印迹法检测2组细胞中EZH2mRNA及蛋白的表达差异,CCK-8方法及Transwell培养体系检测条件培养基对2组卵巢癌细胞增殖及侵袭能力的影响。结果条件培养基处理后的卵巢癌细胞较普通培养基培养的卵巢癌细胞EZH2表达显著升高,并伴随增殖及侵袭能力的增强;我们通过shRNA沉默卵巢癌细胞中的ezh2基因,建立稳定转染的卵巢癌细胞系,再用条件培养基处理EZH2shRNA稳定转染的卵巢癌细胞,处理后EZH2表达较未转染细胞未见明显升高,说明siRNA成功抑制细胞系中EZH2的表达。CCK-8检测和Transwell侵袭实验结果表明,稳定转染的卵巢癌细胞经条件培养基处理后,脂肪干细胞条件培养基对卵巢癌细胞的增殖、迁移及侵袭能力的促进作用消失。结论人脂肪间充质干细胞可能通过调节卵巢癌细胞中EZH2的表达对卵巢癌细胞的增殖、迁移及侵袭能力起促进作用。
Our previous studies showed that adipose-derived mesenchymal stem cells(ADSCs) could promote the growth and metastasis of ovarian cancer. However, the underlying mechanism was still unclear. The expression of the histone methyltransferase enhancer of zeste homologue 2(EZH2) is often increased in malignant tumours, and EZH2 is involved in the metastasis and growth of ovarian cancer. This study was performed to investigate the role of EZH2 in the promotion of ovarian cancer by ADSCs. Conditionally cultured ovarian cancer cells were transfected with or without EZH2 siRNA, then the mRNA and protein expression of EZH2 were evaluated by qRT-PCR and Western blot assays, respectively;the proliferation, migration and invasion abilities of cancer cells were detected by CCK-8, scratch-wound and Transwell assays, respectively. Data showed that ADSC-conditioned medium significantly increased the proliferation, migrationand invasion abilities of ovarian cancer cells. The mRNA and protein expression levels of EZH2 significantly increased in cells cultured in ADSC-conditioned medium compared with cells cultured in unconditioned medium. Furthermore, inhibition of EZH2 in ovarian cancer cells decreased cell proliferation and invasion even when the cells were cultured with ADSC-conditioned medium(P<0.05). Our study suggested that ADSCs could promote the proliferation,migration and invasion of ovarian cancer cells by increasing the expression of EZH2.
作者
尤敏
王福玲
王雅琦
段莹莹
YOU Min;WANG Fuling;WANG Yaqi;DUAN Yingying(Medical College of Qingdao University, Qingdao 266061, China;Department of Obstetrics and Gynaecology,Affiliated Hospital of Qingdao University, Qingdao 266061, China;Department of Obstetrics and Gynaecology,Yantaishan Hospital, Yantai 264001, China)
出处
《免疫学杂志》
CAS
CSCD
北大核心
2019年第6期499-506,共8页
Immunological Journal
关键词
卵巢癌
脂肪间充质干细胞
EZH2
增殖
侵袭
Ovarian cancer
Adipose-derived mesenchymal stem cells
Enhancer of zeste homologue 2
Proliferation
Metastasis
