【摘要】：目的研究microRNA-211(miR-211)對卵巢癌SKOV-3細胞上皮-間質轉化(EMT)功能的影響及其相關機制。方法 SKOV-3卵巢癌細胞株分別轉染miR-211模擬物(211M組)及其陰性對照模擬物(NCM組),并設立未轉染對照組,采用RT-PCR法檢測各組細胞miR-211含量;Transwell實驗檢測3組細胞遷移能力和侵襲能力;Western blot法檢測3組細胞Snail、α-連環(huán)蛋白(α-Catenin)和與性別決定相關的高遷移率基因群4(SOX4)表達水平;采用Western blot法檢測SOX4過表達對miR-211抑制EMT的拮抗作用;雙熒光素酶實驗檢測miR-211與SOX4的關系。結果 211M組miR-211的表達水平明顯上調,表達水平為未轉染對照組的(706.67±30.95)倍(P0.05)。211M組遷移細胞數量為(12.32±0.77)個/視野,明顯低于未轉染對照組的(82.25±1.05)個/視野(P0.05)。211M組侵襲細胞數量為(9.22±0.32)個/視野,明顯低于未轉染對照組的(62.10±1.77)個/視野(P0.05)。211M組細胞Snail蛋白表達量明顯降低,α-Catenin蛋白表達量明顯升高,SOX4蛋白表達量明顯降低。SOX4+211M組卵巢癌細胞中Snail蛋白表達量明顯升高,α-Catenin蛋白表達量明顯降低。雙熒光素酶檢驗結果顯示SOX4為miR-211的下游靶基因。結論 miR-211可能通過降低下游靶基因SOX4水平影響EMT相關蛋白表達,抑制卵巢癌SKOV-3細胞的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】： 中山大學附屬第一醫(yī)院婦產科;
【基金】：國家自然科學基金青年科學基金項目(編號:81602723) 廣東省科技發(fā)展專項(公益研究與能力建設)(編號:2016A020215214) 廣東省醫(yī)學科研基金資助項目(編號:A2015130)
【分類號】：R737.31
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本文編號：2419950