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磷脂酶 scramblase 1 通过 IL-6/JAK/STAT3 通路促进神经胶质瘤的恶性进展。

Phospholipid scramblase 1 acts through the IL-6/JAK/STAT3 pathway to promote the malignant progression of glioma.

机构信息

Department of Neurosurgery, Shandong Cancer Hospital and Institute, Shandong First Medical University and Shandong Academy of Medical Sciences, Jinan, Shandong, 250117, China.

出版信息

Arch Biochem Biophys. 2024 Jun;756:110002. doi: 10.1016/j.abb.2024.110002. Epub 2024 Apr 16.

DOI:10.1016/j.abb.2024.110002

PMID:38636689

Abstract

BACKGROUND

Phospholipid scramblase 1 (PLSCR1) is a calcium-dependent endofacial plasma-membrane protein that plays an essential role in multiple human cancers. However, little is known about its role in glioma. This study aimed to investigate PLSCR1 function in glioma, and elucidate its underlying molecular mechanisms.

METHODS

PLSCR1 expression in human glioma cell lines (U87MG, U251, LN229, A172 and T98G) and human astrocytes was detected by western blot and qRT-PCR. PLSCR1 was silenced using si-PLSCR1-1 and si-PLSCR1-2 in LN229 and U251 cells. PLSCR1 was overexpressed using the pcDNA-PLSCR1 plasmid in T98G cells. Colony formation, 5-ethynyl-2'-deoxyuridine, flow cytometry and transwell assays were employed for measuring cell proliferation, apoptosis and mobility after PLSCR1 knockdown or overexpression. PLSCR1 function in glycolysis in glioma cells was determined through measuring the extracellular acidification rate, oxygen consumption rate, glucose consumption and lactate production. Besides, immunohistochemistry, western blot and qRT-PCR were utilized to assess mRNA and protein expression. Besides, the effect of PLSCR1 silencing on subcutaneous tumor was also monitored.

RESULTS

PLSCR1 expression was upregulated in glioma. The downregulation of PLSCR1 repressed the proliferation, mobility, epithelial-to-mesenchymal transition (EMT) and glycolysis; however, it facilitated apoptosis in glioma cells. Whereas, PLSCR1 upregulation had the opposite effect. Moreover, PLSCR1 promoted the activation of the IL-6/JAK/STAT3 pathway in glioma cells. Besides, IL-6 treatment significantly reversed the function of PLSCR1 silencing on cell proliferation, mobility, EMT, apoptosis and glycolysis. In a nude mouse tumor model, silencing PLSCR1 suppressed tumor growth via inactivating IL-6/JAK/STAT3 signaling.

CONCLUSION

Our results indicated that PLSCR1 could facilitate proliferation, mobility, EMT and glycolysis, but repress apoptosis through activating IL-6/JAK/STAT3 signaling in glioma. Therefore, PLSCR1 may function as a potential therapeutic target for glioma.

摘要

背景

磷脂翻转酶 1（PLSCR1）是一种钙依赖性的内侧面质膜蛋白，在多种人类癌症中发挥着重要作用。然而，关于其在神经胶质瘤中的作用知之甚少。本研究旨在探讨 PLSCR1 在神经胶质瘤中的功能，并阐明其潜在的分子机制。

方法

通过 Western blot 和 qRT-PCR 检测人神经胶质瘤细胞系（U87MG、U251、LN229、A172 和 T98G）和人星形胶质细胞中 PLSCR1 的表达。使用 si-PLSCR1-1 和 si-PLSCR1-2 在 LN229 和 U251 细胞中沉默 PLSCR1。使用 pcDNA-PLSCR1 质粒在 T98G 细胞中过表达 PLSCR1。在 PLSCR1 敲低或过表达后，通过集落形成、5-乙炔基-2'-脱氧尿苷、流式细胞术和 Transwell 测定法测量细胞增殖、凋亡和迁移。通过测量细胞外酸化率、耗氧量、葡萄糖消耗和乳酸产生来确定 PLSCR1 在神经胶质瘤细胞糖酵解中的作用。此外，还利用免疫组织化学、Western blot 和 qRT-PCR 评估 mRNA 和蛋白表达。此外，还监测了 PLSCR1 沉默对皮下肿瘤的影响。

结果

PLSCR1 在神经胶质瘤中表达上调。PLSCR1 的下调抑制了神经胶质瘤细胞的增殖、迁移、上皮间质转化（EMT）和糖酵解，但促进了细胞凋亡。相反，PLSCR1 的上调则产生相反的效果。此外，PLSCR1 促进了神经胶质瘤细胞中 IL-6/JAK/STAT3 通路的激活。此外，IL-6 处理显著逆转了 PLSCR1 沉默对细胞增殖、迁移、EMT、凋亡和糖酵解的作用。在裸鼠肿瘤模型中，沉默 PLSCR1 通过抑制 IL-6/JAK/STAT3 信号通路抑制肿瘤生长。