一种新型多环螺环氧化吲哚类化合物对肺癌细胞系的生长抑制作用及其机制

Inhibitory Effect and Mechanism of a Novel Polycyclic Spiro-epoxy Indole Compound on Lung Cell Lines Proliferation

关键词:  多环螺环氧化吲哚  SKLU1  RhoGEF7  IL-6/JAK/STAT3  NF-κB激活

Key words:  polycyclic spiro-epoxy indole  SKLU1  RhoGEF7  IL-6/JAK/STAT3  NF-κB activation

摘要:

目的 探究一种新型多环螺环氧化吲哚化合物对肺细胞系生长抑制的作用与机制。方法 首先通过MTT试验、流式细胞技术与细胞划痕试验检测化合物对肺癌细胞系的增殖、细胞凋亡以及细胞迁移的影响。接着通过PharmMapper平台预测其靶标蛋白，通过GEPIA平台预测与该靶蛋白具有较强相关性的蛋白，最后分别通过双荧光素酶报告基因试验，qPCR试验以及免疫印迹试验，在启动子水平、转录水平和表达水平检测化合物对目的基因的影响。结果 实验结果表明化合物能够显著抑制SKLU1细胞系的增殖，IC₅₀值为11.38 μmol·L^-1，化合物浓度>2.85 μmol·L^-1即能够抑制SKLU1的细胞迁移，浓度>5.69 μmol·L^-1能够诱导细胞凋亡。其分子机理可能是通过靶向RhoGEF7蛋白，下调下游的Rac1蛋白表达量，从而下调E-cadherin蛋白表达量，抑制细胞迁移；并且，下调的Rac1蛋白还能抑制NF-κB的激活，下调下游基因IL-6和IL-8的转录；下调的Rac1蛋白还能抑制IL-6/JAK/STAT3通路。结论 本研究为这种新型多环螺环氧化吲哚化合物靶向肺癌细胞的分子机制提供理论依据。

Abstract:

OBJECTIVE To explore the effect and mechanism of a novel polycyclic spiro-epoxy indole compound on suppression of lung cell lines proliferation. METHODS The compound’s effect on cellular proliferation, apoptosis, and cellular migration was determined using the MTT assays, flow cytometry, and cell scratch test. The target proteins of this compound were then predicted using the PharmMapper platform, while the associated proteins in lung adenocarcinoma were predicted using the GEPIA platform. In order to determine the effect of this compound on the target gene at the promoter level, transcription level, and expression level, reporter gene assay, qPCR assay and Western blotting test were also performed. RESULTS The results showed that this compound inhibited the proliferation of SKLU1 cell line with an IC₅₀ value of 11.38 μmol·L^-1, inhibited the cell migration of SKLU1 above the concentration of 2.85 μmol·L^-1, and induced the apoptosis above the concentration of 5.69 μmol·L^-1. Additionally, this compound might target the RhoGEF7 and inhibit the cellular migration by down-regulating the expression of Rac1 protein and E-cadherin protein. Then the down-regulated Rac1 protein suppressed the activation of NF-κB, which reduced the transcription of downstream genes IL-6 and IL-8. Moreover, the IL-6/JAK/STAT3 pathway was inhibited when Rac1 protein was down-regulated. CONCLUSION This study provides theoretical basis for the molecular mechanism of targeting lung cancer cells with this novel polycyclic spiro epoxidized indole compound.