Department of Clinical Laboratory, Shenzhen Longhua District Central Hospital, Guangdong Medical University, Shenzhen, 518110, China.
Laboratory Medicine, Guangdong Medical University, Zhanjiang, 524023, China.
Cell Biochem Biophys. 2023 Dec;81(4):727-735. doi: 10.1007/s12013-023-01166-9. Epub 2023 Sep 2.
NLRP12 can affect the progression of different diseases, including hepatocellular carcinoma. However, no report on triple-negative breast cancer (TNBC) has been found. Thus, this study aimed to explore the role of NLRP12 in TNBC. In our study, immunohistochemistry, real-time quantitative PCR (qPCR), and Western blot assays were used to evaluate NLRP12 expression in TNBC tissues and cells. Then, NLRP12 lentivirus was constructed and infected into MDA-MB-231 and MDA-MB-157 cells with or without PTD-p65-P1 treatment. Next, cells were collected for cell function detection using the following procedures: colony formation assay for proliferation, Transwell for migration and invasion, and Western blot for NF-κB and MAPK pathway-associated proteins. Finally, a xenograft mouse model was applied; the tumor volume and weight were determined, and NLRP12, p-IκBb-α, and p-IκBb-α expressions were evaluated using qPCR and Western blot. Results indicated that NLRP12 was lowly expressed in TNBC tissues and cells. The inhibition of NLRP12 could induce the proliferation, migration, and invasion of TNBC cells, which also could be reversed by inhibiting the NF-κB pathway (PTD-p65-P1). Moreover, silencing of NLRP12 could upregulate p-IκBb-α, while IκBb-α, p-ERK, ERK, p-p38, p38, p-JNK, and JNK expressions remained unchanged, thereby indicating that only the NF-κB pathway could be activated by NLRP12 silencing. Furthermore, the xenograft mouse model confirmed the abovementioned findings. Therefore, the low expression of NLRP12 promoted the proliferation, migration, and invasion in TNBC cells by activating the NF-κB pathway. This study might provide insights into TNBC therapy.
NLRP12 可以影响包括肝细胞癌在内的不同疾病的进展。然而,尚未发现关于三阴性乳腺癌(TNBC)的报道。因此,本研究旨在探讨 NLRP12 在 TNBC 中的作用。在我们的研究中,使用免疫组织化学、实时定量 PCR(qPCR)和 Western blot 检测 NLRP12 在 TNBC 组织和细胞中的表达。然后,构建 NLRP12 慢病毒并感染 MDA-MB-231 和 MDA-MB-157 细胞,同时或不进行 PTD-p65-P1 处理。接下来,通过集落形成实验、Transwell 实验和 Western blot 实验检测细胞功能,分别用于增殖、迁移和侵袭的检测,以及 NF-κB 和 MAPK 通路相关蛋白的检测。最后,应用异种移植小鼠模型;测定肿瘤体积和重量,并通过 qPCR 和 Western blot 评估 NLRP12、p-IκBb-α 和 p-IκBb-α 的表达。结果表明,NLRP12 在 TNBC 组织和细胞中低表达。抑制 NLRP12 可诱导 TNBC 细胞的增殖、迁移和侵袭,而抑制 NF-κB 通路(PTD-p65-P1)可以逆转这一现象。此外,沉默 NLRP12 可上调 p-IκBb-α,而 IκBb-α、p-ERK、ERK、p-p38、p38、p-JNK 和 JNK 的表达保持不变,这表明只有 NF-κB 通路可以被 NLRP12 沉默激活。此外,异种移植小鼠模型证实了上述发现。因此,NLRP12 的低表达通过激活 NF-κB 通路促进 TNBC 细胞的增殖、迁移和侵袭。本研究可能为 TNBC 治疗提供新的思路。