| 周美娟,黄海波,林志祥,丁振华.紫外线B辐射敏感的miR-365在皮肤鳞癌发生中的作用研究[J].中华放射医学与防护杂志,2014,34(11):813-816,866 |
| 紫外线B辐射敏感的miR-365在皮肤鳞癌发生中的作用研究 |
| The carcinogenic effect of UVB sensitive miR-365 in cutaneous squamous cell carcinoma |
| 投稿时间：2014-04-30 |
| DOI：10.3760/cma.j.issn.0254-5098.2014.11.004 |
| 中文关键词: miR-365 紫外线B 皮肤鳞状细胞癌 癌基因 |
| 英文关键词:MiR-365 UVB Cutaneous squamous cell carcinoma Oncogene |
| 基金项目:国家自然科学基金(81172634,81202152) |
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| 中文摘要: |
| 目的 探讨紫外线(UVB)辐射敏感miR-365在皮肤鳞状细胞癌发生中的作用.方法 取对数生长期正常人皮肤角质细胞HaCaT分为对照组和照射组,照射组给予50 J/m2的UVB照射.照射后培养不同时间,分析miRNA的表达谱.实时荧光定量PCR分析HaCaT细胞和皮肤鳞癌细胞系A431、Tca8113和HSC-1的miR-365的表达.平板克隆形成实验和Transwell小室细胞运动实验分别检测细胞的克隆形成能力和体外运动能力.构建miR-365高表达的HaCaT细胞系HaCaTpre-miR-365-2.裸鼠皮下注射HaCaTpre-miR-365-2观察其成瘤能力.结果 UVB照射后,HaCaT差异表达的miRNAs共30个,其中miR-365最敏感,且照后6 h升高最明显,为对照的6.7倍;皮肤鳞癌细胞系A431、Tca8113和HSC-1的miR-365的表达均高于HaCaT细胞(P<0.05),其中,A431增加最多,为(15.67±1.12)倍,Tca8113最少,为(4.72±0.85)倍;抑制皮肤鳞癌细胞系miR-365的表达可以显著抑制其克隆形成能力(t=13.68,P<0.05)和体外细胞迁移能力(t=19.98,P<0.05),而提高HaCaT的miR-365的表达则可以显著提高其克隆形成能力(t=7.11,P<0.05)和体外细胞迁移能力(t=22.03,P<0.05),且能够在裸鼠皮下成功地成瘤.结论 miR-365是一种皮肤鳞状细胞癌的促癌基因. |
| 英文摘要: |
| Objective To investigate the carcinogeic role of miR-365 in cuntanerous squamous cell carcinoma (cSCC). Methods Normal HaCaT cells were divided into control and irradiation groups, the later was exposed by UVB irradiation (50 J/m2). MicroRNA expression profiles of the two groups were analyzed by microRNA array. The expression variations of miR-365 in HaCaT, A431, Tca8113 and HSC-1 cells were validated by qRT-PCR analysis. The colony-forming and invasion capacities were dectected by colony forming assay and Transwell migration assay in vitro, respectively. HaCaTpre-miR-365-2 highly expressing miR-365 was constructed by retroviral vector infection. Tumorigenicity evaluation was carried out by subcutaneously inject of the cells at the right back flank of nude mice. Results There were 30 microRNAs differentially expressed in HaCaT cells after UVB irradiation and miR-365 was one of the most sensitive miRNAs(as high 6.7 times as control). Expression of miR-365 in all the cSCC cell lines A431, Tca8113 and HSC-1 were significantly higher than that in HaCaT cell, in which the maximum was A431 (15.67±1.12 times,P<0.01), and the minimum was Tca8113(4.72±0.85 times,P<0.05). Knockdown of miR-365 in cSCC cell lines significantly inhibited the colony forming ability (t=13.68,P<0.05) and cell migration (t=19.98,P<0.05) in vitro. HaCaT cells overexpressing miR-365 by transient transfection significantly increased the ability of colony formation (t=7.11,P<0.05) and cell migration (t=22.03,P<0.05) in vitro. In addition, HaCaTpre-miR-365-2 cell line stably expressing miR-365 could successfully establish tumors in nude mice. Conclusions MiR-365 is an oncogene for cutaneous squamous cell carcinoma. |
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