| 引用本文： |
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杨波,温晓飞,刘辉,等.miR-1721靶向CD44抑制前列腺癌PC-3细胞增殖、侵袭和肿瘤干细胞形成的研究[J].同济大学学报（医学版）,2016,37(4):25-30. [点击复制]
- YANG Bo,WEN Xiao-fei,LIU Hui,et al.miR-1721 suppresses proliferation and invasion of prostate cancer cells and its stem cells by targeting CD44[J].Journal of Tongji University(Medical Science),2016,37(4):25-30. [点击复制]
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| 摘要: |
| 目的 探讨miR-1721靶向抑制CD44的表达及对前列腺癌细胞增殖、侵袭能力的影响。方法 构建CD44 mRNA 3′-非翻译区(3′-UTR)的荧光素酶报告载体系统,通过荧光素酶系统确定miR-1721对CD44 mRNA 3′-UTR的靶向关系；脂质体转染法将miR-1721模拟物转入前列腺癌PC-3细胞中,Western印迹法检测转染后CD44蛋白表达水平的改变；MTT法检测miR-1721mimics对PC-3细胞增殖能力的影响,Transwell小室法检测其对肿瘤细胞袭侵袭能力的影响；通过干细胞成球实验检测miR-1721mimics对前列腺癌干细胞样微球体形成的影响。结果 miR-1721能特异性地与CD44 mRNA 3′-UTR结合,抑制其荧光素酶的活性,干细胞标记基因CD44是miR-1721的靶基因。过表达miR-1721的PC-3细胞中CD44蛋白表达水平明显降低,可显著抑制PC-3细胞的侵袭能力,同时过表达CD44可负调控miR-1721对PC-3细胞增殖和侵袭能力的影响；干细胞成球实验证实,miR-1721能显著抑制前列腺癌细胞PC-3干细胞的微球体形成能力。结论 miR-1721通过靶向调控CD44的表达而抑制前列腺癌细胞增殖、侵袭能力及干细胞的形成能力。 |
| 关键词: 前列腺肿瘤 CD44 干细胞标记基因 miRNAs |
| DOI：10.16118/j.1008-0392.2016.04.005 |
| 通信作者: |
| 投稿时间：2015-12-04 |
| 录用日期： |
| 基金项目:上海市卫生和计划生育委员会科研课题(20134428)；上海市浦东新区卫生系统优秀学科带头人培养计划(PWRd2011-08)；吴阶平医学基金(320.6750.14197) |
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| miR-1721 suppresses proliferation and invasion of prostate cancer cells and its stem cells by targeting CD44 |
| YANG Bo,WEN Xiao-fei,LIU Hui,LIU Feng,DENG Xiao-jun,LIAO Guo-qiang |
| (Dept.of Urology, Shanghai Zhoupu Hospital, Shanghai University of Medicine & Health Sciences, Shanghai 201318, China;Dept.of Ulology, East Hospital, Tongji University, Shanghai 200120, China) |
| Abstract: |
| Objective To investigate the effect of miR-1721 on the proliferation and invasion of prostate cancer cells by targeting CD44. Methods The CD44 3′-untranslated region mRNA-luciferase reporter vector was constructed and the dual-luciferase reporter gene assay was employed to examine the effect of miR-1721 on activity of luciferase. Prostate cancer PC-3 cells were transfected with miR-1721 mimics by LipofectAMINE-2000, and the expressions level of CD44 protein was detected by Western blotting. The inhibition effects of CD44 on cell proliferation and invasion were observed after CD44 siRNA were transfected into PC-3 cells. PC-3-sc cells mammosphere assay was performed after cotransfection with miR-1721 mimics and CD44 siRNA. Results miR-1721 could bind to the 3′-UTR of CD44 and inhibited the activity of luciferase. Then CD44 protein expression was significantly down-regulated when miR-1721 was overexpressed in PC-3 cells. Overexpression of miR-1721 inhibited the invasion of PC-3 cells. Overexpression of miR-1721 antagonized a role of proliferation and invasion in PC-3 cells. Overexpression of miR-1721 reduced mammosphere number and the size of prostate cancer stem cells. Conclusion The miR-1721 expression can suppress cell proliferation and invasion by targeting CD44 in prostate cancer cells. |
| Key words: prostate carcinoma CD44 stem cell genetic marker miRNAs |
