Department of Oncology, Affiliated Hospital of Chengdu University of Traditional Chinese Medicine. No. 39 Twelve Bridges Road, Jinniu District, Chengdu 610072, China.
Department of Oncology, Sichuan Integrative Medicine Hospital. No. 51, Fourth Section of Renmin South Road, Chengdu 610041, China.
Biosci Rep. 2020 Jul 31;40(7). doi: 10.1042/BSR20200730.
Gemcitabine is widely used in the treatment of breast cancer (BC). However, the resistance to drugs remains a tough concern. The study explored the potential mechanism concerning gemcitabine resistance in triple-negative BC (TNBC) in vitro.
TNBC cells (TNBCC) and gemcitabine-resistance cell lines (GRC) were used. We investigated the sensitivity to gemcitabine responsive to regulation of Nod-like receptor protein 3 (NLRP3) expression in TNBCC in different gemcitabine concentrations. RT-PCR checked NLRP3 mRNA expression and MTT assessed the cell cytotoxicity. Gemcitabine resistance was studied in GRC exposed to 0, 1, 3, 5 nm gemcitabine after GRC were treated with NLRP3 agonist Nigericin sodium salt (NSS) or antagonist CY-09. Epithelial-to-mesenchymal transition (EMT) biomarkers were evaluated via RT-PCR and inflammasome IL-1β, β-catenin content and GSK-3β activity were measured by ELISA methods. Last, we inactivated the signaling and examined the NLRP3, EMT mRNA expression by RT-PCR, IL-1β, β-catenin content and GSK-3β activity by ELISA and cell cytotoxicity through MTT.
NLRP3 up-regulation improved cell survival and reduced sensitivity to gemcitabine (P<0.05). NLRP3 had higher expression in GRC than TNBCC. GRC cell viability dropped as the gemcitabine concentration increased. NLRP3 up-regulation added to resistance to gemcitabine in GRC (P<0.05). NLRP3 agonist might induce EMT process, activate wnt/β-catenin signaling and IL-1β, while inactivation of wnt/β-catenin signaling could result in the inhibition of NLRP3, IL-1β and EMT as well as cell viability in GRC (P<0.05).
NLRP3 could enhance resistance to gemcitabine via IL-1β/EMT/Wnt/β-catenin signaling, which suggested that NLRP3 antagonist CY-09 might be incorporated into gemcitabine treatment for TNBC.
吉西他滨被广泛应用于乳腺癌（BC）的治疗。然而，药物耐药仍然是一个严峻的问题。本研究旨在探讨体外三阴性乳腺癌（TNBC）中吉西他滨耐药的潜在机制。
使用 TNBC 细胞（TNBCC）和吉西他滨耐药细胞系（GRC）。我们研究了不同吉西他滨浓度下 TNBCC 中 NLRP3 表达调节对吉西他滨敏感性的影响。RT-PCR 检测 NLRP3 mRNA 表达，MTT 评估细胞细胞毒性。用 NLRP3 激动剂 Nigericin 钠盐（NSS）或拮抗剂 CY-09 处理 GRC 后，研究 GRC 暴露于 0、1、3、5nm 吉西他滨时的吉西他滨耐药性。通过 RT-PCR 评估上皮间质转化（EMT）标志物，通过 ELISA 方法测量炎症小体 IL-1β、β-catenin 含量和 GSK-3β 活性。最后，我们抑制信号通路，通过 RT-PCR 检测 NLRP3、EMT mRNA 表达，通过 ELISA 检测 IL-1β、β-catenin 含量和 GSK-3β 活性，通过 MTT 检测细胞活力。
NLRP3 上调可提高细胞存活率，降低吉西他滨敏感性（P<0.05）。GRC 中的 NLRP3 表达高于 TNBCC。随着吉西他滨浓度的增加，GRC 细胞活力下降。NLRP3 上调增加了 GRC 对吉西他滨的耐药性（P<0.05）。NLRP3 激动剂可能诱导 EMT 过程，激活 wnt/β-catenin 信号和 IL-1β，而抑制 wnt/β-catenin 信号可抑制 GRC 中的 NLRP3、IL-1β 和 EMT 以及细胞活力（P<0.05）。
NLRP3 可能通过 IL-1β/EMT/Wnt/β-catenin 信号增强吉西他滨耐药性，提示 NLRP3 拮抗剂 CY-09 可能被纳入 TNBC 的吉西他滨治疗。
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