The Second Affiliated Hospital of Shandong First Medical University, Tai'an 271000, Shandong, China.
Taian Maternal and Child Health Hospital, Tai'an 271000, Shandong, China.
J Healthc Eng. 2021 Jul 23;2021:4650794. doi: 10.1155/2021/4650794. eCollection 2021.
The vast majority of primary hepatocellular cancer is hepatocellular carcinomas (HCCs). Currently, HCC is one of the more common cancers in humans, and it has a high mortality and disability rate. Mitoxantrone (MTX) is an antitumor drug that can block type II topoisomerase. It has been reported that immunogenic cell death evoked by MTX can induce the discharge of damage associated with molecular patterns (DAMPs) and subsequently influence immune cell infiltration in the tumor microenvironment. High mobilities aggregation box 1 (HMGB1) is the prototypical extracellular DAMP. Many cellular processes have been reported to involve EPHB4 receptor tyrosine kinases, but the relation of DAMP and EPHB4 is uncertain. In this research, we assessed the impact of miR-130-3p by Edu incorporation test on cell proliferation, and we have proven its impact on HCC cell migration through Transwell and wound healing tests. Flow cytometry was applied to study its influence on apoptosis. Luciferase report test was integrated in detecting the miR-130-3p target gene. The influence of miR-130-3p on the manifestation of classical DAMPs was studied, such as HMGB1, ATP, and Calreticulin. A coculture experiment was carried out to further confirm its effects on immune cell infiltration. The result displayed that miR-130-3p overexpression considerably facilitates apoptosis and suppresses the migration or proliferation of HCC cells. EPHB4 was confirmed as the target gene of miR-130-3p. Overexpression of this target gene promotes emission of Calreticulin, ATP, and HMGB1 and subsequently promotes DCs maturation and proliferation of CD4+ T cells. In summary, our results demonstrated that miR-130-3p inhibits HCC cell proliferation and migration by targeting EPHB4 and promotes drug-induced immunogenic cell death.
绝大多数原发性肝癌是肝细胞癌（HCC）。目前，HCC 是人类中较为常见的癌症之一，具有较高的死亡率和致残率。米托蒽醌（MTX）是一种抗肿瘤药物，可阻断 II 型拓扑异构酶。据报道，MTX 诱导的免疫原性细胞死亡可引发损伤相关分子模式（DAMPs）的释放，进而影响肿瘤微环境中免疫细胞的浸润。高迁移率族蛋白 B1（HMGB1）是典型的细胞外 DAMPs。据报道，许多细胞过程都涉及 EPHB4 受体酪氨酸激酶，但 DAMPs 与 EPHB4 的关系尚不确定。在这项研究中，我们通过 Edu 掺入试验评估了 miR-130-3p 对细胞增殖的影响，并通过 Transwell 和划痕愈合试验证明了其对 HCC 细胞迁移的影响。流式细胞术用于研究其对细胞凋亡的影响。荧光素酶报告试验用于检测 miR-130-3p 的靶基因。研究了 miR-130-3p 对经典 DAMPs（如 HMGB1、ATP 和钙网蛋白）表达的影响。进行共培养实验以进一步确认其对免疫细胞浸润的影响。结果显示，miR-130-3p 的过表达可显著促进细胞凋亡，并抑制 HCC 细胞的迁移或增殖。EPHB4 被确认为 miR-130-3p 的靶基因。该靶基因的过表达可促进钙网蛋白、ATP 和 HMGB1 的释放，进而促进 DC 成熟和 CD4+T 细胞的增殖。总之，我们的结果表明，miR-130-3p 通过靶向 EPHB4 抑制 HCC 细胞的增殖和迁移，并促进药物诱导的免疫原性细胞死亡。
