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目的 探讨淫羊藿苷（ICA）通过调控长链非编码RNA(lncRNA)OIP5-AS1/miR-338-3p通路对肝癌细胞增殖、迁移及侵袭的影响。方法 采用四甲基偶氮唑蓝（MTT）法检测ICA对肝癌SNU182细胞增殖的抑制作用，并筛选合适的干预浓度。将肝癌SNU182细胞分为Control组、ICA组、ICA+阴性对照（pc-NC）组、ICA+OIP5-AS1过表达（pc-OIP5-AS1）组、ICA+pc-OIP5-AS1+阴性对照（miR-NC）组和ICA+pc-OIP5-AS1+miR-338-3p模拟物（miR-338-3p mimics）组。采用实时荧光定量PCR检测OIP5-AS1和miR-338-3p表达水平；MTT法检测细胞增殖能力；细胞划痕实验检测细胞迁移能力；Transwell实验检测细胞侵袭能力；流式细胞术检测细胞凋亡水平；蛋白质免疫印迹法检测血管内皮生长因子-A(VEGF-A)、血管内皮钙黏蛋白（VE-cadherin）、增殖细胞核抗原（PCNA）、裂解的半胱氨酸蛋白酶3(cleaved caspase-3)、上皮钙黏蛋白（E-cadherin）、神经钙黏蛋白（N-cadherin）及波形蛋白（Vimentin）的表达水平；双荧光素酶活性实验验证OIP5-AS1和miR-338-3p的靶向关系。结果 12.5～100μmol/L的ICA均可抑制SNU182细胞增殖（P<0.05）；选取50μmol/L的ICA进行后续实验。与Control组比较，ICA组SNU182细胞的OIP5-AS1表达水平、生长活性、划痕愈合率和侵袭数量、N-cadherin、Vimentin、VEGF-A、VE-cadherin和PCNA蛋白表达水平降低，而miR-338-3p表达水平、凋亡率、E-cadherin和cleaved caspase-3蛋白表达水平升高（P<0.05）。与ICA组和ICA+pc-NC组比较，ICA+pc-OIP5-AS1组SNU182细胞的OIP5-AS1表达水平、生长活性、划痕愈合率和侵袭数量、N-cadherin、Vimentin、VEGF-A、VE-cadherin和PCNA蛋白表达水平升高，而miR-338-3p表达水平、凋亡率、E-cadherin和cleaved caspase-3蛋白表达水平降低（P<0.05）；而进一步上调miR-338-3p表达可逆转过表达OIP5-AS1对SNU182细胞恶性生物学行为的促进作用（P<0.05）。结论 ICA可能通过下调OIP5-AS1和上调miR-338-3的表达，抑制SNU182细胞增殖、迁移和侵袭，并促进细胞凋亡。
Abstract：Objective To explore the impacts of icariin(ICA) on the proliferation, migration, and invasion of liver cancer cells by adjusting long non-coding RNA(lncRNA) OIP5-AS1/miR-338-3p pathway. Methods MTT assay was used to measure the inhibitory effect of ICA on the proliferation of liver cancer SNU182 cells, and screen for the appropriate intervention concentration. Liver cancer SNU182 cells were classified into Control group, ICA group, ICA+pc-NC group, ICA+pc-OIP5-AS1 group, ICA+pc-OIP5-AS1+miR-NC group, and ICA+pc-OIP5-AS1+miR-338-3p mimics group. Real-time quantitative PCR was performed to measure the expression of OIP5-AS1 and miR-338-3p. MTT assay was used to detect cell proliferation. Cell scratch assay was used to detect cell migration. Transwell method was used to measure cell invasion. Flow cytometry was used to measure cell apoptosis. Western blot was performed to detect the protein expression of vascular endothelial growth factor A(VEGF-A), vascular endothelial cadherin(VE-cadherin), proliferating cell nuclear antigen(PCNA), cleaved caspase-3, E-cadherin, N-cadherin and vimentin. The dual luciferase activity assay verified the targeting relationship between OIP5-AS1 and miR-338-3p. Results 12.5-100 μmol/L of ICA could inhibit the proliferation of SNU182 cells(P<0.05), and 50 μmol/L was selected for the subsequent experiments. Compared with the Control group, the OIP5-AS1 expression, the growth activity, scratch healing rate, invasion number, N-cadherin, vimentin, VEGF-A, VE-cadherin, and PCNA protein expression of SNU182 cells in the ICA group reduced, while the miR-338-3p expression, cell apoptosis rates, E-cadherin and cleaved caspase-3 protein expression increased(P<0.05). Compared with the ICA group and the ICA+pc-NC group, the OIP5-AS1 expression, the growth activity, scratch healing rate, invasion number, N-cadherin, vimentin, VEGF-A, VE-cadherin, and PCNA protein expression in SNU182 cells in the ICA+pc-OIP5-AS1 group increased, while the miR-338-3p expression, cell apoptosis rates, E-cadherin and cleaved caspase-3 protein expression reduced(P<0.05). Upregulation of miR-338-3p could reduce the promoting effect of overexpression of OIP5-AS1 on the malignant biological behavior of SNU182 cells(P<0.05). Conclusion ICA may inhibit the proliferation, migration and invasion of SNU182 cells while promoting apoptosis by downregulating the expression of OIP5-AS1 and upregulating the expression of miR-338-3p.
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基本信息:
中图分类号:R285
引用信息:
[1]郝亚明,顾秀锋,龙志雄.淫羊藿苷通过调控lncRNA OIP5-AS1/miR-338-3p通路抑制肝癌细胞的增殖、迁移与侵袭[J].临床肿瘤学杂志,2026,31(02):127-133.