miR-21抑制黑色素瘤细胞凋亡的机制研究  

Mechanism of miR-21 inhibiting apoptosis of melanoma cells

作　　者：周启明[1,2] 李巍[3] 袁建辉[4] 赵艳[1,2] 段江曼[1,2] 熊东林[1,2] 

机构地区：[1]深圳市南山区人民医院,广东深圳518052 [2]深圳市疼痛学重点实验室 [3]深圳市第三人民医院 [4]深圳市疾病预防控制中心

出　　处：《华南预防医学》2017年第3期235-239,共5页South China Journal of Preventive Medicine

基　　金：深圳市知识创新计划基础研究项目(JCY20140411092959833);深圳市科技计划项目(ZDSYS20140509150415945);深圳市科技创新委员会基础研究学科布局项目(JCYJ20160328161613864)

摘　　要：目的验证黑色素瘤患者与正常人血清中miR-21水平差异,明确其在黑色素瘤细胞凋亡中的作用,为探索黑色素瘤潜在的治疗靶标提供理论基础。方法分别采集黑色素瘤患者与正常人血清样本,提取microRNA后利用qRT-PCR分析miR-21在血清中的水平,同时采用ELISA法检测其靶基因PDCD4在血清中的含量。构建miR-21稳定抑制的黑色素瘤细胞株,进一步分析其对PDCD4表达的影响。采用流式细胞术分析miR-21对黑色素瘤细胞凋亡的影响。结果 miR-21在黑色素瘤患者血清中的表达是阴性对照组的(4.62±2.42)倍,其靶基因PDCD4表达是阴性对照组的(0.36±0.21)倍,差异均具有统计学意义(均P<0.01)。miR-21在黑色素瘤细胞中表达被抑制时,PDCD4在A-375和SK-MEL-1细胞中的表达分别是阴性对照组的(1.69±0.39)和(2.20±1.06)倍,细胞凋亡水平分别是空白质粒对照组的(2.82±0.24)、(2.26±0.23)倍,差异均具有统计学意义(均P<0.01)。结论 miR-21在黑色素瘤患者血清中异常升高,推测其能够抑制凋亡蛋白相关蛋白PDCD4的表达,进一步抑制黑色素瘤细胞凋亡,提示miR-21可能作为黑色素瘤治疗的潜在靶分子。Objective To verify the difference of serum miR-21 levels between patients with melanoma and healthy persons, clarify its role in the apoptosis of melanoma cells, and explore the potential targets for melanoma therapy. Methods Sera were collected from patients with melanoma and healthy control to extract microRNAs. RT-PCR was used to quantify the relative serum level of miR-21. The target gene programmed cell death-4 （PDCD4） was also analyzed as the potential target of miR-21 by ELISA. The miR-21 inhibitory melanoma cells was constructed and its effect on PDCD4 expression was identified. The effect of miR-21 on apoptosis was analyzed by flow-cytometric assay. Results Compared with the negative control, the level of miR-21 in sera of melanoma patients was up-regulated （4.62±2.42 ） and its potential target PDCD4 was down-regulated （0.36±0.21 ） （ both P 〈 0.01 ）. As the expression of miR-21 was in- hibited in melanoma cells, the expressions of PDCD4 in A-375 and SK-MEL-1 cells were （ 1.69 ＋ 0.39 ） and （2.20±1.06） times of the negative control group respectively, and the cell apoptosis was increased in miR-21 over expressed melanoma cells compared with negative control （A-375 ： 2, 82±0.24, SK-MEL-1 ： 2.26±0.23 ） through target PDCD4 （ P 〈 0.01 ）. Conclusion miR-21 was increased in patients with melanoma and might inhibit tumor cell apoptosis by inducing down-regulation of PDCD4 in melanoma cells,suggesting that miR-21 may be a potential target in melanoma therapy.

关 键 词：黑色素瘤 微RNAS 细胞凋亡 

分 类 号：R739.5[医药卫生—肿瘤]