【摘要】：目的探討microRNA-205(miR-205)表達(dá)對(duì)人類黑素瘤細(xì)胞株(A375和A2058)遷移和侵襲的作用及其分子機(jī)制。方法轉(zhuǎn)染miR-205過(guò)表達(dá)慢病毒(Lenti-miR-205)構(gòu)建miR-205過(guò)表達(dá)的黑素瘤細(xì)胞A375和A2058細(xì)胞系(miR-205組),轉(zhuǎn)染空白對(duì)照慢病毒作為對(duì)照細(xì)胞系(NC組)。通過(guò)劃痕實(shí)驗(yàn)和Transwell實(shí)驗(yàn)分別檢測(cè)兩組細(xì)胞遷移和侵襲能力,激光共聚焦掃描顯微鏡觀察細(xì)胞形態(tài)和表達(dá)鈣黏蛋白E(E-cadherin)的細(xì)胞數(shù)量與熒光強(qiáng)度,蛋白質(zhì)印跡法檢測(cè)E-cadherin、Twist、整合素(intergrin)、波形蛋白(vimentin)、Zeb1、基質(zhì)金屬蛋白酶(MMP)-2、MMP-9蛋白表達(dá)。結(jié)果培養(yǎng)8 h和24 h時(shí)miR-205組A375與A2058細(xì)胞的遷移能力均大于NC組(P0.01);miR-205組兩種細(xì)胞的侵襲能力均低于NC組(P0.01)。MiR-205過(guò)表達(dá)后A375和A2058細(xì)胞均由梭形、基質(zhì)型向圓形、表皮型轉(zhuǎn)化。與NC組相比,miR-205組細(xì)胞E-cadherin蛋白表達(dá)增加(P0.01),且表達(dá)E-cadherin的細(xì)胞數(shù)量和熒光強(qiáng)度均增加;Twist、intergrin、vimentin、Zeb1、MMP-2和MMP-9蛋白表達(dá)水平降低(P0.01)。結(jié)論 MiR-205通過(guò)誘導(dǎo)E-cadherin的表達(dá)逆轉(zhuǎn)上皮間質(zhì)轉(zhuǎn)化,從而抑制黑素瘤細(xì)胞的遷移和侵襲。
[Abstract]:Objective to investigate the effect of microRNA-205 (miR-205) expression on migration and invasion of human melanoma cell lines (A375 and A2058) and its molecular mechanism. Methods miR-205 overexpression lentivirus (Lenti-miR-205) was transfected into A375 and A2058 melanoma cells with miR-205 overexpression (miR-205 group), and blank control lentivirus was transfected as control cell line (NC group). The migration and invasion ability of the two groups were detected by scratch test and Transwell assay. The cell number and fluorescence intensity of the cells expressing cadherin E (E-cadherin) and morphology were observed by confocal laser scanning microscopy. The expression of E-cadherin Twist, integrin (intergrin), vimentin (vimentin) Zeb1, matrix metalloproteinase (MMP)-2 and MMP-9 were detected by Western blot. Results the migration ability of A375 and A2058 cells in miR-205 group was higher than that in NC group (P0.01) at 8 h and 24 h. The invasion ability of A375 and A2058 cells in miR-205 group was lower than that in NC group (P0.01). MiR-205 overexpression was lower than that in NC group (P0.01). Compared with NC group, the expression of E-cadherin protein was increased (P0.01), and the number and fluorescence intensity of E-cadherin were increased (P0.01). The expression of MMP-2 and MMP-9 protein were decreased (P0.01). Conclusion MiR-205 inhibits the migration and invasion of melanoma cells by inducing E-cadherin expression to reverse epithelial mesenchymal transformation.
【作者單位】： 復(fù)旦大學(xué)附屬華山醫(yī)院皮膚科;
【基金】：上海市衛(wèi)生局科研課題(20134363)~~
【分類號(hào)】：R739.5

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