本文選題：miR-a　切入點(diǎn)：葡萄膜黑色素瘤　出處：《眼科新進(jìn)展》2017年07期

【摘要】：目的分析miR26a在葡萄膜黑色素瘤中的表達(dá)及對葡萄膜黑色素瘤細(xì)胞增殖、凋亡、遷移及侵襲的影響。方法對葡萄膜黑色素瘤細(xì)胞系SP6.5、M23及正常葡萄膜上皮細(xì)胞系A(chǔ)RPE-19行RT-PCR實(shí)驗(yàn),檢測3種細(xì)胞系中miR-26a的相對表達(dá)差異;將葡萄膜黑色素瘤細(xì)胞系SP6.5分成miR-26a模擬物組和NC組,分別轉(zhuǎn)染miR-26a mimics和對照序列scramble,用CCK-8實(shí)驗(yàn)和流式細(xì)胞術(shù)分別檢測兩組細(xì)胞增殖和凋亡;采用細(xì)胞劃痕實(shí)驗(yàn)及Transwell實(shí)驗(yàn)分別檢測兩組細(xì)胞的遷移和侵襲能力;采用Western blot檢測其下游蛋白組蛋白甲基化轉(zhuǎn)移酶果蠅zeste基因增強(qiáng)子同源物2(enhancer of zeste homolog 2,EZH2)的表達(dá)水平。結(jié)果葡萄膜黑色素瘤細(xì)胞系SP6.5中miR-26a相對表達(dá)量為0.250±0.029,細(xì)胞系M23中為0.350±0.017,正常葡萄膜上皮細(xì)胞系A(chǔ)RPE-19中miR-26a相對表達(dá)量為1.0,在細(xì)胞系SP6.5及M23中miR-26a的相對表達(dá)量均低于正常葡萄膜上皮細(xì)胞系A(chǔ)RPE-19中miR-26a的相對表達(dá)量(均為P0.001)。miR-26a模擬物組在培養(yǎng)72 h、96 h、120 h OD450值(0.69±0.09、1.23±0.15、2.12±0.23)均顯著低于NC組(1.39±0.11、2.35±0.25、3.53±0.27),差異均有統(tǒng)計(jì)學(xué)意義(均為P0.05)。miR-26a模擬物組細(xì)胞凋亡率(15.60±2.30)%高于NC組(5.00±0.70)%(P0.01);miR-26a模擬物組劃痕愈合率(23.7±2.1)%低于NC組(68.9±5.1)%(P0.01);侵襲細(xì)胞數(shù)(45.1±3.9)個低于NC組(115.3±8.9)個(P0.01)。miR-26a模擬物組EZH2蛋白相對表達(dá)量(0.39±0.09)低于NC組(1.0;P0.01)。結(jié)論 miR-26a在葡萄膜黑色素瘤中低表達(dá),過表達(dá)miR-26a顯著抑制葡萄膜黑色素瘤細(xì)胞增殖,促進(jìn)凋亡,并抑制遷移和侵襲,其可能是通過下調(diào)EZH2的表達(dá)來發(fā)揮作用的。
[Abstract]:Objective to investigate the expression of miR26a in uveal melanoma and its effect on the proliferation, apoptosis, migration and invasion of uveal melanoma cells.Methods RT-PCR assay was performed on the uveal melanoma cell line SP6.5 M23 and the normal uveal epithelial cell line ARPE-19 to detect the relative expression of miR-26a in the three cell lines, and the SP6.5 cell line was divided into miR-26a mimic group and NC group.The proliferation and apoptosis of miR-26a mimics and control sequence were detected by CCK-8 assay and flow cytometry respectively, and the migration and invasion ability of the two groups were detected by cell scratch test and Transwell assay, respectively.Western blot was used to detect the expression of the enhancer 2(enhancer of zeste homolog 2 (EZH2) in Drosophila melanogaster zeste gene.Results the relative expression of miR-26a was 0.250 鹵0.029 in uveal melanoma cell line SP6.5, 0.350 鹵0.017 in M23 cell line, and 1.0 in normal uveal epithelial cell line ARPE-19. The relative expression of miR-26a in SP6.5 and M23 cell lines was lower than that in normal staphylococcal cell line.钀勮啘涓婄毊緇嗚優(yōu)緋籄RPE-19涓璵iR-26a鐨勭浉瀵硅〃杈鵑噺(鍧囦負(fù)P0.001).miR-26a妯℃嫙鐗╃粍鍦ㄥ煿鍏，

本文編號：1702701