背景:龙胆。是一种传统的藏药,具有镇痛和抗炎活性。环状RNA(circularRNAs,circRNAs)已被确定参与非小细胞肺癌(NSCLC)的肿瘤发生。这里,本研究旨在探讨龙胆黄酮(GF)和circ_0059665在NSCLC进展中的作用和机制。
方法:采用qRT-PCR和蛋白质印迹分析检测mRNA和蛋白的含量。细胞增殖和侵袭能力通过细胞计数试剂盒-8,EdU,集落形成和transwell分析,分别。通过流式细胞术分析M2巨噬细胞极化。
结果:GF治疗抑制NSCLC细胞增殖,低氧条件下的侵袭和M2巨噬细胞极化。Circ_0059665在NSCLC组织和细胞中高表达。其表达在低氧条件下增加,但在GF处理后降低。此外,circ_0059665过表达逆转了低氧条件下GF对NSCLC细胞的抗癌作用。机械上,circ_0059665充当miR-512-5p的海绵来调节NOVA2表达。缺氧降低miR-512-5p水平,NSCLC细胞中NOVA2水平升高,而这些趋势在GF治疗后被消除。Circ_0059665沉默抑制NSCLC细胞增殖,低氧环境中的侵袭和M2巨噬细胞极化,被NOVA2过表达抵消。此外,NOVA2上调逆转了GF对低氧处理NSCLC细胞的抑制作用。此外,GF通过抑制circ_0059665阻碍体内NSCLC肿瘤生长。
结论:GF治疗在低氧环境中抑制NSCLC细胞增殖,侵袭和M2巨噬细胞极化通过circ_0059665/miR-512-5p/NOVA2轴。
BACKGROUND: Gentiana macrophylla Pall. is a traditional Tibetan medicinal herb possessing antinociceptive and anti-inflammatory activities. Circular RNAs (circRNAs) have been identified to be involved in the tumorigenesis of non-small cell lung cancer (NSCLC). Here, this study focused on investigating the function and mechanism of Gentiana macrophylla flavonoids (
GF) and circ_0059665 in NSCLC progression.
METHODS: The contents of mRNA and protein were detected using qRT-PCR and western blotting analysis. Cell proliferative and invasive abilities were evaluated by cell counting kit-8, EdU, colony formation and transwell assays, respectively. M2 macrophage polarization was analyzed by flow cytometry.
RESULTS: GF treatment suppressed NSCLC cell proliferation, invasion and M2 macrophage polarization under hypoxic conditions. Circ_0059665 was highly expressed in NSCLC tissues and cells. Its expression was increased under hypoxic conditions but was reduced following
GF treatment. Furthermore, circ_0059665 overexpression reversed the anticancer effects of GF on NSCLC cells under hypoxic conditions. Mechanistically, circ_0059665 acted as a sponge for miR-512-5p to regulate NOVA2 expression. Hypoxia decreased miR-512-5p levels, and increased NOVA2 levels in NSCLC cells, while these tendencies were abolished after
GF treatment. Circ_0059665 silencing inhibited NSCLC cell proliferation, invasion and M2 macrophage polarization in hypoxic environments, which were counteracted by NOVA2 overexpression. Moreover, NOVA2 upregulation reversed the suppressive effects of
GF on NSCLC cells with hypoxia treatment. In addition,
GF impeded NSCLC tumor growth in vivo via suppressing circ_0059665.
CONCLUSIONS: GF treatment in hypoxic environments suppressed NSCLC cell proliferation, invasion and M2 macrophage polarization via the circ_0059665/miR-512-5p/NOVA2 axis.