Major of integrated Chinese and Western Medicine, Guangzhou University of Chinese Medicine, Guangzhou, Guangdong, China.
Department of Thyroid Surgery, Linyi People's Hospital, Linyi, Shandong, China.
Biotechnol Genet Eng Rev. 2024 Oct;40(2):1389-1405. doi: 10.1080/02648725.2023.2193775. Epub 2023 Mar 22.
Triptolide (TP) is involved in the progression of liver cancer. However, the detailed molecular network regulated through TP is still unclear. Long non-coding RNA (LncRNA) SLC9A3 exerts roles in various pathological progresses. Nevertheless, whether SLC9A3 affects the sensitivity of liver cancer cells to TP have not been uncovered. The content of SLC9A3-AS1 and miR-449b-5p was estimated by utilizing quantitative real-time polymerase-chain reaction (qRT-PCR). Cell counting kit 8 (CCK-8) assay was introduced to assess cell viability. Additionally, cell viability as well as invasion was tested via transwell assay. The direct binding between miR-449b-5p and SLC9A3-AS1 or LDHA was confirmed through luciferase reporter gene assay. Moreover, glycolysis rate was tested by calculating the uptake of glucose in addition to the production of lactate in Huh7 cells. LncRNA SLC9A3-AS1 was up-regulated in liver cancer tissue samples and cells. Knockdown of SLC9A3-AS1 notably further inhibited viability, migration as well as invasion in Huh7 cells. MiR-449b-5p was the direct downstream miRNA of SLC9A3-AS1 and was down-regulated by SLC9A3-AS1 in Huh7 cells. In addition, miR-449b-5p was reduced in liver cancer tissues and cells. Overexpressed miR-449b-5p increased the sensitivity of Huh7 cells to TP remarkably. Moreover, miR-449b-5p negatively regulated LDHA expression in Huh7 cells. This work proved that SLC9A3-AS1 increased the sensitivity of liver cancer cells to TP by regulating glycolysis rate mediated via miR-449b-5p/LDHA axis. These findings implied that TP is likely to be a potent agent for treating patients diagnosed with liver cancer.
雷公藤红素(TP)参与肝癌的进展。然而,TP 调节的详细分子网络仍不清楚。长链非编码 RNA(LncRNA)SLC9A3 在各种病理进展中发挥作用。然而,SLC9A3 是否影响肝癌细胞对 TP 的敏感性尚未被揭示。通过实时定量聚合酶链反应(qRT-PCR)测定 SLC9A3-AS1 和 miR-449b-5p 的含量。细胞计数试剂盒 8(CCK-8)测定法用于评估细胞活力。此外,通过 Transwell 测定法检测细胞活力和侵袭能力。通过荧光素酶报告基因测定证实了 miR-449b-5p 与 SLC9A3-AS1 或 LDHA 之间的直接结合。此外,通过计算 Huh7 细胞中葡萄糖的摄取以及乳酸的产生来测试糖酵解率。LncRNA SLC9A3-AS1 在肝癌组织样本和细胞中上调。SLC9A3-AS1 的敲低显着进一步抑制 Huh7 细胞的活力、迁移和侵袭。MiR-449b-5p 是 SLC9A3-AS1 的直接下游 miRNA,在 Huh7 细胞中受 SLC9A3-AS1 下调。此外,miR-449b-5p 在肝癌组织和细胞中减少。过表达 miR-449b-5p 显着增加 Huh7 细胞对 TP 的敏感性。此外,miR-449b-5p 负调节 Huh7 细胞中的 LDHA 表达。这项工作证明,SLC9A3-AS1 通过调节 miR-449b-5p/LDHA 轴介导的糖酵解率增加肝癌细胞对 TP 的敏感性。这些发现表明,TP 可能是治疗肝癌患者的有效药物。