摘要
目的:探讨miR-155在转化生长因子-β1(transforming growth factor-β1,TGF-β1)诱导的人前列腺癌细胞PC3中的生物学作用。方法:TGF-β1处理PC3细胞,相差倒置显微镜下观察细胞形态学变化;通过Transwell试验检测细胞的迁移能力;实时荧光定量PCR(quantitative real-time polymerasechain reaction,qRT-PCR)和蛋白质印迹法检测TGF-β1诱导的PC3细胞纤维连接蛋白(fibronectin,FN)的表达情况;qRT-PCR检测miR-155的表达水平;干扰miR-155检测TGF-β1诱导的PC3细胞FN的表达及细胞迁移能力的变化。结果:TGF-β1诱导PC3细胞发生形态改变并增强其迁移能力,且FN的表达显著上调;TGF-β1诱导的miR-155表达呈现时间依赖性;干扰miR-155使TGF-β1诱导的FN表达减少,细胞形态及迁移能力在一定程度上发生逆转。结论:miR-155在TGF-β1诱导的前列腺癌细胞PC3中高表达,可能间接上调FN的表达从而促进细胞迁移,可为前列腺癌的治疗提供理论依据。
Objective: To investigate the biological roles of miR-155 in transforming growth factor-β1 (TGF-β1)-treated human prostate cancer cells PC3. Methods: PC3 cells were treated with TGF-β1. The morphological changes were observed under phase-contrast microscopy. Migration of PC3 cells was detected by Transwell assay. The expression of FN was assessed by quantitative real-time polymerase chain reaction (qRT-PCR), Western blotting and immunofluorescence staining. miR-155 treated by TGF-β1 were analyzed by qRT-PCR. After interfering with miR-155 in TGF-β1-treated PC3 cells, qRT-PCR and Western blotting were used to detect FN expression. Migration of TGF-β1-treated PC3 cells interfered with miR-155 was detected by Transwell assay. Results:TGF-β1 treatment could induce morphological alteration of PC3 cells from compact, cobblestone morphology into spindle-shaped, fibroblast-like morphology. Transwell assay showed that TGF-β1 significantly increased the migration of PC3 cells. qRT-PCR, Western blotting and immunofluorescence staining demonstrated that TGF-β1 up-regulated the expression of FN. The expression of miR-155 induced by TGF-β1 was in a time-dependent manner. Introduction of miR-155 inhibitor partly attenuated the expression of FN in mRNA and protein levels, and inhibited the change of cell morphology and migration in TGF-β1 induced PC3 cells. Conclusion: miR-155 is up-regulated by TGF-β1 in PC3 cells, which may affect cell migration by increasing FN expression. It may have implications for treatment of human prostate cancer.
作者
曾珊珊
李盼
刘浩
陈丹扬
ZENG Shanshan;LI Pan;LIU Hao;CHEN Danyang(Cancer Research Institute, Affiliated Cancer Hospital & Guangzhou Medical University,Guangzhou Key Laboratory of Translational Medicine on Cancer Treatment,Guangzhou 510095,China)
出处
《临床与病理杂志》
2019年第9期1865-1871,共7页
Journal of Clinical and Pathological Research
基金
广东省自然科学基金(2017A030313500)
广州市科技计划一般项目(201707010381)
广州市属高校科研项目(1201630143)~~
