目的：探讨长链非编码RNA锌指结构反义转录本1（lncRNA ZFAS1）调控miR-588/高迁移率蛋白A2（HMGA2）轴对肝癌HepG2细胞增殖、侵袭、迁移的影响。方法：qPCR、WB法检测80例肝癌组织及对应癌旁组织（2018年1月至2019年12月在武汉第三医院首义院区手术切除标本）及人正常 肝 LO2 细 胞 和 肝 癌 HepG2、Huh7、HCCLM3 细胞中 ZFAS1、miR-588 及HMGA2表达水平；采用Kaplan-Meier进行患者生存曲线分析。将 HepG2 细胞分为空白组、si-NC组、si-ZFAS1组、si-ZFAS1+inhibitor NC组、si-ZFAS1+miR-588 inhibitor组；qPCR检测各组HepG2细胞中ZFAS1、miR-588表达水平，WB法检测各组HepG2细胞中HMGA2蛋白表达；CCK-8法、Transwell和划痕实验分别检测各组HepG2细胞的增殖、侵袭和迁移能力；双荧光素酶报告基因实验分别验证ZFAS1和miR-588、miR-588和HMGA2的靶向关系。用HepG2细胞移植瘤裸鼠模型检测敲减ZFAS1或/和miR-588 对移植瘤生长的影响。结果:在肝癌组织和肝癌细胞中，ZFAS1、HMGA2 呈高表达，miR-588 呈低表达（均 P<0.05）；ZFAS1低表达患者2年生存率高于高表达组（P<0.05）；与空白组比较 ，si-ZFAS1 组 ZFAS1、HMGA2表达水平显著降低，miR588表达水平显著升高（均P<0.05）。与si-ZFAS1组比较，si-ZFAS1+miR-588 inhibitor组中ZFAS1表达水平无显著变化（P>0.05），HMGA2表达水平显著升高、miR-588表达水平显著降低（P<0.05）；敲减ZFAS1可抑制HepG2细胞的增殖、侵袭和迁移能力，并抑制裸鼠体内移植瘤的生长（均P<0.05）；ZFAS1靶向miR-588并抑制后者的表达，miR-588靶向HMGA2并抑制后者的表达；同时抑制ZFAS1和miR-588表达可逆转敲减ZFAS1对HepG2细胞增殖、侵袭与迁移能力及体内移植瘤生长的抑制作用（均P<0.05）。结论:敲减ZFAS1可通过促进miR-588表达来下调HMGA2表达，进而抑制肝癌HepG2细胞的增殖、侵袭与迁移能力。

Objective: To investigate the effects of long non-coding RNA zinc finger antisense 1 (lncRNA ZFAS1) on the proliferation, invasion, and migration of liver cancer cells by regulating the miR-588/high mobility group AT-hook protein 2 (HMGA2) axis. Methods: Real-time fluorescent quantitative PCR (qRT-PCR) and western blot were performed to measure the expression levels of ZFAS1, miR-588 and HMGA2 in 80 pairs of liver cancer tissues and corresponding para-cancerous issues (the tissue sample were collected from Shouyi Campus, Wuhan Third Hospital during Jan. 2018 and Dec. 2019), human normal liver cell line (LO2) and liver cancer cell lines (HepG2, Huh7, HCCLM3). The survival of patients was analyzed using the Kaplan-Meier survival curve. HepG2 cells were divided into blank group, si-NC group, si-ZFAS1 group, si-ZFAS1+inhibitor NC group, and si-ZFAS1+miR-588 inhibitor group. qPCR was performed to measure the expression of ZFAS1 and miR-588 in HepG2 cells of each group, and Western blot was performed to measure the expression of HMGA2 protein in the cells. CCK-8 method, Transwell, and scratch test were performed to measure the proliferation, invasion, and migration of HepG2 cells. Dual-luciferase reporter gene experiment was performed to verify the targeting relationship between ZFAS1 and miR-588 as well as between miR-588 and HMGA2. A HepG2 cell transplanted tumor model was established in nude mice to examine the effect of silencing ZFAS1 or/and miR-588 on the growth of transplanted tumors. Results: ZFAS1 and HMGA2 were highly expressed while miR-588 was lowly expressed in liver cancer tissues and liver cancer cells (all P<0.05). The 2-year survival rate of patients with low ZFAS1 expression was higher than that in the high expression group (P<0.05). Compared with the blank group, the relative expression of ZFAS1 and HMGA2 protein in the si-ZFAS1 group was significantly reduced, and the relative expression of miR-588 was significantly increased (all P<0.05); compared with the si-ZFAS1 group, the relative expression of ZFAS1 in the si-ZFAS1+miR-588 inhibitor group did not change significantly (P>0.05), however, the relative expression of HMGA2 protein was significantly increased, and the relative expression of miR-588 was significantly reduced (P<0.05). Silencing ZFAS1 was able to inhibit the proliferation, invasion, and migration of HepG2 cells and inhibit the growth of transplanted tumors in nude mice (all P<0.05). ZFAS1 targeted and down-regulated the expression of miR-588, while miR-588 targeted and down-regulated the expression of HMGA2. Simultaneous inhibition of miR-588 expression could reverse the inhibitory effects of silencing ZFAS1 on the proliferation, invasion, and migration of HepG2 cells and the growth of transplanted tumors in nude mice (all P<0.05). Conclusion: Silencing ZFAS1 may down-regulate the expression of HMGA2 by promoting the expression of miR-588, thereby inhibiting the proliferation, invasion, and migration of liver cancer HepG2 cells.

R735.7; R730.2

武汉市卫生和计划生育委员会科研项目资助（No.WX18Q17）