【摘要】：目的研究microRNA-211(miR-211)對(duì)卵巢癌SKOV-3細(xì)胞上皮-間質(zhì)轉(zhuǎn)化(EMT)功能的影響及其相關(guān)機(jī)制。方法 SKOV-3卵巢癌細(xì)胞株分別轉(zhuǎn)染miR-211模擬物(211M組)及其陰性對(duì)照模擬物(NCM組),并設(shè)立未轉(zhuǎn)染對(duì)照組,采用RT-PCR法檢測(cè)各組細(xì)胞miR-211含量;Transwell實(shí)驗(yàn)檢測(cè)3組細(xì)胞遷移能力和侵襲能力;Western blot法檢測(cè)3組細(xì)胞Snail、α-連環(huán)蛋白(α-Catenin)和與性別決定相關(guān)的高遷移率基因群4(SOX4)表達(dá)水平;采用Western blot法檢測(cè)SOX4過表達(dá)對(duì)miR-211抑制EMT的拮抗作用;雙熒光素酶實(shí)驗(yàn)檢測(cè)miR-211與SOX4的關(guān)系。結(jié)果 211M組miR-211的表達(dá)水平明顯上調(diào),表達(dá)水平為未轉(zhuǎn)染對(duì)照組的(706.67±30.95)倍(P0.05)。211M組遷移細(xì)胞數(shù)量為(12.32±0.77)個(gè)/視野,明顯低于未轉(zhuǎn)染對(duì)照組的(82.25±1.05)個(gè)/視野(P0.05)。211M組侵襲細(xì)胞數(shù)量為(9.22±0.32)個(gè)/視野,明顯低于未轉(zhuǎn)染對(duì)照組的(62.10±1.77)個(gè)/視野(P0.05)。211M組細(xì)胞Snail蛋白表達(dá)量明顯降低,α-Catenin蛋白表達(dá)量明顯升高,SOX4蛋白表達(dá)量明顯降低。SOX4+211M組卵巢癌細(xì)胞中Snail蛋白表達(dá)量明顯升高,α-Catenin蛋白表達(dá)量明顯降低。雙熒光素酶檢驗(yàn)結(jié)果顯示SOX4為miR-211的下游靶基因。結(jié)論 miR-211可能通過降低下游靶基因SOX4水平影響EMT相關(guān)蛋白表達(dá),抑制卵巢癌SKOV-3細(xì)胞的EMT功能。
[Abstract]:Objective to study the effect of microRNA-211 (miR-211) on epithelial-interstitial transformation of SKOV-3 cells and its related mechanism. Methods SKOV-3 ovarian cancer cell lines were transfected with miR-211 analogue (211M group) and negative control group (NCM group), and the untransfected control group was set up. The miR-211 content of each group was detected by RT-PCR assay. The expression levels of Snail, 偽-catenin (偽-Catenin) and high mobility gene group 4 (SOX4) were detected by Transwell assay and; Western blot assay. Western blot assay was used to detect the antagonistic effect of SOX4 overexpression on miR-211 inhibiting EMT, and double luciferase assay was used to detect the relationship between miR-211 and SOX4. Results the expression level of miR-211 in 211M group was (706.67 鹵30.95) times higher than that in untransfected control group (P0.05), and the number of migration cells in 211M group was (12.32 鹵0.77) / visual field. The number of invasive cells in 211M group was (9.22 鹵0.32) / visual field, which was significantly lower than that in untransfected control group (82.25 鹵1.05) / visual field (P0.05). The expression of Snail protein in 211M group was significantly lower than that in untransfected control group (62.10 鹵1.77) / visual field (P0.05). In SOX4 211M group, the expression of Snail protein was significantly increased, and the expression of 偽 -Catenin protein was significantly decreased. Double luciferase assay showed that SOX4 was the downstream target gene of miR-211. Conclusion miR-211 inhibits the EMT function of ovarian cancer SKOV-3 cells by decreasing the SOX4 level of the downstream target gene and affecting the expression of EMT related proteins.
【作者單位】： 中山大學(xué)附屬第一醫(yī)院婦產(chǎn)科;
【基金】：國(guó)家自然科學(xué)基金青年科學(xué)基金項(xiàng)目(編號(hào):81602723) 廣東省科技發(fā)展專項(xiàng)(公益研究與能力建設(shè))(編號(hào):2016A020215214) 廣東省醫(yī)學(xué)科研基金資助項(xiàng)目(編號(hào):A2015130)
【分類號(hào)】：R737.31
，

本文編號(hào)：2419950