miR-130b通过调控PTEN在人视网膜母细胞瘤发挥促癌作用

发布时间：2018-07-08 08:17

本文选题：视网膜母细胞瘤 + miR-b　；参考：《眼科》2017年04期

【摘要】：目的探讨mi R-130b在人视网膜母细胞瘤(retinoblastoma,RB)中的作用,并阐明其分子机制。设计实验研究。研究对象人RB癌组织及细胞系。方法采用荧光定量PCR检测40例RB患者癌组织及相应癌旁组织中mi R-130b的表达情况。采用噻唑蓝比色法(MTT)和蛋白印迹法检测mi R-130b对RB细胞增殖及凋亡能力的影响。通过生物信息学分析预测mi R-130b的靶基因,并利用双荧光素酶报告基因实验和蛋白印迹验证。主要指标细胞生存率、细胞凋亡率及荧光素酶活性。结果mi R-130b在人RB癌组织的表达量明显高于配对的癌旁组织(P=0.023);mi R-130b在HXO-Rb44(P=0.008)和Y79(P=0.012)细胞系中的表达量亦高于正常视网膜血管内皮细胞(ACBRI-181)。体外功能实验发现,转染mi R-130b抑制物可显著抑制HXO-Rb44(P=0.004)和Y79(P=0.001)细胞的增殖,并增加HXO-Rb44(P=0.011)和Y79(P=0.027)细胞凋亡。生物信息学分析发现"与张力蛋白同源10号染色体缺失的磷酸酶"(phosphatase and tensin homologue deleted on chromosome 10,PTEN)是mi R-130b的靶基因。双荧光素酶报告基因试验和蛋白印迹结果显示,mi R-130b可直接与PTEN m NRA 3’-非编码区结合,并抑制其翻译,并最终降低HXO-Rb44(P=0.038)和Y79(P=0.025)细胞中PTEN蛋白表达水平。结论 mi R-130b在RB组织表达上调,mi R-130b通过下调PTEN的表达可促进RB细胞的增殖并减少其凋亡,提示mi R-130b可作为RB基因治疗的一个新的分子靶点。
[Abstract]:Objective to investigate the role of mi R-130b in human retinoblastoma (RB) and to elucidate its molecular mechanism. Design experimental research. Human RB cancer tissues and cell lines were studied. Methods the expression of miR-130b in 40 patients with RB and its adjacent tissues was detected by fluorescence quantitative PCR. The effects of miR-130b on the proliferation and apoptosis of RB cells were detected by MTT and Western blot. The target gene of mi R-130b was predicted by bioinformatics analysis, and the double luciferase reporter gene experiment and Western blotting were used. Main outcome measures cell survival rate apoptosis rate and luciferase activity. Results the expression of miR-130b in human RB cancer tissues was significantly higher than that in matched paracancerous tissues (P0. 023). The expression of miR-130b in HXO-Rb44 (P0. 008) and Y79 (P0. 012) cell lines was also higher than that in normal retinal vascular endothelial cells (ACBRI-181). In vitro functional experiments showed that the transfection of mi R-130b inhibitor significantly inhibited the proliferation of HXO-Rb44 (P0. 004) and Y79 (P0. 001) cells, and increased the apoptosis of HXO-Rb44 (P0. 011) and Y79 (P0. 027) cells. Bioinformatics analysis showed that "phosphatase" (phosphatase and tensin homologue deleted on chromosome 10) was the target gene of mi R-130b. The results of double luciferase reporter gene assay and Western blotting showed that PTEN R-130b could bind directly to PTEN m NRA3- non-coding region, inhibit its translation, and ultimately reduce the expression of PTEN protein in HXO-Rb44 (Pt0.038) and Y79 (Pn0.025) cells. Conclusion the up-regulation of miR-130b expression in RB can promote the proliferation of RB cells and decrease its apoptosis by down-regulating the expression of PTEN, suggesting that miR-130b may be a new molecular target for RB gene therapy.
【作者单位】：河南省濮阳市油田总医院眼科;
【分类号】：R739.7

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