目的：探究瑞戈非尼（regorafenib，Rego）对人肝癌SMMC-7721细胞增殖、凋亡的影响及其可能的机制。方法：将SMMC-7721细胞分为对照组及 Rego 组 ，分别用 0、10 μmol/L Rego 处理 48 h 后 ，流式细胞术检测各组细胞凋亡率 ，qPCR检测细胞中 miR-122 的表达。采用脂质体转染的方法将合成的hsa-miR-122-5p模拟物转染SMMC-7721细胞构建miR-122过表达的 overExp-miR-122 细胞，并将细胞分为对照组、Rego 组、overExp-NC 组、overExp-NC+Rego 组、overExp-miR-122 组及overExp-miR-122+Rego 组，采用 MTT 法 检 测 细 胞 活 性 ，流 式 细 胞 术 检 测 细 胞 凋 亡 率 、WB 法 检 测 细 胞 中 Bcl2、cleaved caspase-3、RAS、RAF1、p-ERK1蛋白表达水平。结果：与对照组相比，Rego处理后细胞凋亡率显著升高（P<0.05），且miR-122表达量显著上升（P<0.01）；与overExp-NC组比较，overExp-miR-122组细胞增殖抑制率、凋亡率和cleaved caspase-3表达均显著升高（均P<0.01），RAS 蛋白表达显著下降（P<0.05），Bcl2、RAF、p-ERK1 蛋白表达均显著下降（均 P<0.01）；与 overExp-miR-122 组相比，overExp-miR-122+Rego组细胞中各检测指标变化进一步显著增加（P<0.01）。结论：Rego可抑制SMMC-7721细胞增殖、促进凋亡，其作用可能与调控miR-122、凋亡相关因子的表达和抑制RAS/RAF/ERK信号通路有关。

Objective: To explore the effects of regorafenib (Rego) on proliferation and apoptosis of human liver cancer SMMC-7721 cells and the possible mechanism. Methods: SMMC-7721 cells were divided into control group and Rego group (10 μmol/L Rego).After the treatment for 48 h, Flow cytometry was used to detect cell apoptosis rate, and qPCR was used to determine the level of miR-122 in two groups of cells. hsa-miR-122-5p mimics were transfected into SMMC-7721 cells by lipofection to construct the miR-122 overexpressing cell line (overExp-miR-122). Then, the cells were divided into control group, Rego group, overExp-NC group,overExp-NC+Rego group, overExp-miR-122 group and overExp-miR-122+Rego group. MTT and Flow cytometry were used to detect cell viability and apoptosis rate, respectively. The protein expression levels of Bcl2, cleaved caspase-3, RAS, RAF1 and p-ERK1 in cells were detected by WB assay. Results: Compared with control group, the cell apoptosis rate was significantly increased after Rego treatment (P<0.05), and miR-122 expression level was significantly increased in Rego group (P<0.01). Compared with overExp-NC group, the proliferation inhibition rate and apoptosis rate in the cells of overExp-miR-122 group were significantly increased (P<0.01),the expression level of cleaved caspase-3 was significantly upregulated, while the protein expressions of Bcl2, RAS, RAF and p-ERK were significantly decreased (P<0.05 or P<0.01). Compared with overExp-miR-122 group, the changes in above detected indicators of overExp-miR-122+Rego group were more obvious (P<0.01). Conclusion: Regorafenib can inhibit the proliferation and promote apoptosis of SMMC-7721 cells, which may be achieved by regulating the expression of miR-122, thereby regulating the expression of apoptosis related factors and inhibiting RAS/RAF/ERK signaling pathway.

湖北省中央引导地方科技发展专项资助（No. 2019ZYYD067）