| 小豆蔻明通过激活NLRP3/Caspase-1信号通路抑制肾癌细胞的侵袭、转移的机制研究 |
| Mechanism of cardamondamin inhibiting invasion and metastasis of renal cancer cells through activation of NLRP3 / Caspase-1 signaling pathway |
| DOI： |
| 中文关键词: 小豆蔻明 NOD样受体蛋白3 胱天蛋白酶-1 肾癌 侵袭转移 细胞焦亡 |
| 英文关键词: Cardamom NOD-like receptor protein 3 Caspase-1 Renal cancer Invasion and metastasis Pyroptosis |
| 基金项目:1.江西省中医药管理局科技计划，编号：2023B0440；2.江西省卫生健康委科技计划，编号：202410252； |
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| 中文摘要: |
| 目的 探讨小豆蔻明通过激活NOD样受体蛋白3(NLRP3)/胱天蛋白酶-1(Caspase-1)信号通路抑制肾癌细胞的侵袭、转移的机制。方法 在肾癌细胞系A498中加入不同浓度的小豆蔻明,利用CCK-8实验观察不同药物浓度下的细胞存活率变化,确定最适加药浓度进行后续实验。将A498细胞分为小豆蔻明组和对照组,利用平板克隆实验、划痕实验、Transwell实验、Westren blot实验[细胞焦亡相关靶标:cleaved-Caspase-1、cleaved-白细胞介素-1β(cleaved-IL-1β)、GSDMD的N端片段(GSDMD-N)]、酶联免疫吸附测定(ELISA)实验(IL-1β及IL18)、实时定量逆转录聚合酶链式反应(qRT-PCR)实验(IL-1β、IL-18的mRNA表达水平)观察细胞的增殖、侵袭、迁移、细胞焦亡的改变。结果 双因素方差分析显示,药物浓度和处理时间对细胞存活率有显著主效应(F浓度=1818.994,F时间=79.405,P<0.05),且存在浓度-时间交互效应(F交互=9.205,P<0.05)；事后分析表明,24h、48h、72h小豆蔻明组的存活率显著低于对照组(P<0.05),当浓度≥20μM时,24h、48h、72h时间点的细胞存活率比较差异具有统计学意义(P<0.05),并呈现剂量-时间-效应关系(P<0.05)。小豆蔻明组细胞克隆数、划痕愈合率、细胞迁移数少于对照组,差异有统计学意义(P<0.05)。小豆蔻明组NLRP3、cleaved-Caspase-1、cleaved-IL-1β、GSDMD-N的表达水平高于对照组,差异有统计学意义(P<0.05)；小豆蔻明组IL-1β及IL-18的表达水平高于对照组,差异有统计学意义(P<0.05)。结论 小豆蔻明可通过激活NLRP3/Caspase-1信号通路促进肿瘤细胞焦亡,抑制肾癌细胞的增殖、侵袭和转移,或可为临床上防治肾癌提供可靠的理论依据。 |
| 英文摘要: |
| Objective To investigate the mechanism of cardamom in inhibiting the invasion and metastasis of renal cancer cells through the activation of NOD-like receptor protein 3 (NLRP3) / caspase-1 (Caspase-1) signaling pathway. Methods Different concentrations of cardamom were added to the renal cancer cell line A498, and CCK-8 was used to determine the optimal drug concentration for subsequent experiments. A498 cells were divided into cardamom group and control group, and the proliferation, invasion, migration and pyroptosis levels were observed by using plate cloning, scratch, Transwell, Westren blot [cleaved-Caspase-1, cleaved-interleukin-1β(cleaved-IL-1β), and N-terminal fragment of the GSDMD (GSDMD-N) ], enzyme-linked immunosorbent assay (ELISA) experiments (IL-1β and IL 18) and qRT-PCR experiments (mRNA expression levels of IL-1β and IL-18). Results A two-way ANOVA showed that, Drug concentration and treatment time had a significant main effect on cell viability (Fconcentration =1818.994, Ftime =79.405, P<0.05), And had a concentration-time interaction effect (F interaction =9.205, P<0.05); Post hoc analysis showed that, The survival rate of 24h, 48h and 72h was significantly lower than the control group (P<0.05), At a concentration of 20 μ M, The difference in cell survival rates at the 24h, 48h and 72h time points was statistically significant (P<0.05), The dose-time-effect relationship was also presented (P<0.05). The number of cell clones, scratch healing rate, and cell migration of the cardamom group were less than that of the control group, and the difference was statistically significant (P<0.05). The expression levels of NLRP3, cleaved-Caspase-1, cleaved-IL-1βand GSDMD-N were higher in cardamom group, statistically significant (P<0.05); the expression levels of IL-1β and IL-18 were higher in cardamom group, statistically significant (P<0.05). Conclusion Cardamom can promote pyroptosis of tumor cells by activating NLRP3/Caspase-1 signaling pathway, inhibit the proliferation, invasion and metastasis of renal cancer cells, or provide a reliable theoretical basis for the prevention and treatment of renal cancer in clinical practice. |
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