目的 探討冬凌草甲素對人肝癌細(xì)胞Huh-7索拉非尼（Sor）耐藥細(xì)胞株Huh-7/Sor的影響及其作用機(jī)制。方法 體外培養(yǎng)Huh-7/Sor細(xì)胞株，qRT-PCR檢測miR-217和ABCC1 mRNA的表達(dá)水平，Western blotting檢測ABCC1蛋白的表達(dá)水平；用冬凌草甲素處理Huh-7/Sor細(xì)胞后，CCK-8法檢測細(xì)胞活力，Annexin V/PI染色檢測細(xì)胞凋亡，Western blotting檢測凋亡蛋白和ABCC1的表達(dá)水平；數(shù)據(jù)庫預(yù)測miR-217與ABCC1的結(jié)合位點，轉(zhuǎn)染miR-217 mimic或inhibitor后檢測ABCC1的蛋白水平，熒光素酶報告基因檢測ABCC1 3’UTR區(qū)域活性。結(jié)果 與Huh-7細(xì)胞相比，Huh-7/Sor細(xì)胞中miR-217的表達(dá)水平顯著下調(diào)，ABCC1的表達(dá)水平顯著上調(diào)（P<0.05）；冬凌草甲素能抑制Huh-7/Sor細(xì)胞的增殖，促進(jìn)細(xì)胞凋亡，上調(diào)miR-217的表達(dá)水平，抑制ABCC1的表達(dá)（P<0.05）；miR-217能與ABCC1 3’UTR區(qū)域結(jié)合，抑制ABCC1的蛋白表達(dá)水平（P<0.05）。結(jié)論 冬凌草甲素能通過上調(diào)miR-217而抑制ABCC1的表達(dá)，進(jìn)而抑制Huh-7/Sor細(xì)胞增殖，誘導(dǎo)凋亡。

Objective To investigate the effect of oridonin on sorafenib resistant human hepatoma cell line Huh-7/Sor and its mechanism. Methods Huh-7/Sor cells were cultured in vitro. The mRNA expression levels of mir-217 and ABCC1 in Huh-7/Sor cells were detected by qRT-PCR and ABCC1 protein was detected by Western blotting. After treated with oridonin, the cell viability was detected by CCK-8 method, apoptosis was detected by Annexin V/PI staining, and the protein expression of apoptotic proteins and ABCC1 were detected by Western blotting. The binding sites of miR-217 and ABCC1 were predicted by database. The protein level of ABCC1 was detected by Western blotting after transfection with miR-217 mimic or inhibitor. The activity of ABCC1 3' UTR region was detected by luciferase reporter gene. Results Compared with Huh-7 cells, the expression level of miR-217 in Huh-7/Sor was down-regulated, and the expression level of ABCC1 was significantly up-regulated (P<0.05). Oridonin inhibited the proliferation of Huh-7/Sor cells and promoted apoptosis. The expression of miR-217 was up-regulated and the expression of ABCC1was down-regulated in Huh-7/Sor cells treated by Oridonin. miR-217 could bind with ABCC1 3' UTR region and inhibit the protein expression of ABCC1 (P<0.05). Conclusion Oridonin could inhibit the proliferation of Huh-7/Sor cells and induce apoptosis by up regulating the expression of ABCC1 and miR-217.

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