Zhang Yu, Chen Weipeng, Cheng Xin, Wang Feiran, Gao Cheng, Song Fei, Song Fengliang, Liang Xiaoliang, Fang Wanzhi, Chen Zhong
Department of Hepatobiliary Surgery, Affiliated Hospital of Nantong University, Nantong, China.
School of Medicine, Nantong University, Nantong, China.
Front Oncol. 2022 May 24;12:852765. doi: 10.3389/fonc.2022.852765. eCollection 2022.
Hepatocellular carcinoma (HCC) is one of the most common malignant tumors worldwide, with unclear pathogenesis. Sphingomyelin phodiesterase acid-like 3A (SMPDL3A) affects cell differentiation and participates in immune regulation. However, its molecular biological function in HCC has not yet been elucidated.
Data from 180 HCC patients were analyzed the relationship between the expression of SMPDL3A in liver cancer tissues and the prognosis of liver cancer patients. Crispr-Cas9 dual vector lentivirus was used to knock out SMPDL3A in HCC cell lines. The effects of SMPDL3A on cell viability were determined by CCK8 assay, clone formation experiment, cell cycle assay, cell scratch, TUNEL experiment and flow cytometry. Xenograft tumor assays in BALB/c nude mice confirmed that SMPDL3A promoted tumor growth and . Preliminary exploration of SMPDL3A interacting protein by mass spectrometry analysis and co-immunoprecipitation.
This study showed that the expression of SMPDL3A in HCC tissue differed from that in tumor-adjacent tissues. Moreover, the overall survival rate and tumor-free survival rate of patients with high-SMPDL3A expression were significantly lower than those with low-SMPDL3A expression. SMPDL3A expression was closely related to the level of protein induced by PIVKA-II, liver cirrhosis, tumor diameter, microvascular invasion, and Barcelona clinic liver cancer staging. Thus, SMPDL3A is an independent risk factor that affects the tumor-free survival rate and overall survival rate of HCC patients. study using Crispr-Cas9 genome editing technology revealed the knockout effect of on cell proliferation, apoptosis, and migration. Cell counting kit-8 assay and clone formation experiment showed that sgSMPDL3A inhibited tumor cell proliferation and migration. Flow cytometry and TUNEL assay showed that sgSMPDL3A promoted apoptosis in tumors. Moreover, sgSMPDL3A inhibited tumor growth during subcutaneous tumor formation in nude mice. Immunohistochemistry of Ki67 and PNCA also indicated that sgSMPDL3A inhibited subcutaneous tumor proliferation in tumor-bearing nude mice. Further experiments showed that SMPDL3A interacts with the enhancer of rudimentary homolog (ERH).
High-SMPDL3A expression was related to poor prognosis of patients with HCC. Knockout of inhibited the proliferation and migration and accelerated the migration of HCC cells. SMPDL3A interacted with ERH to affect the tumorigenesis and progression of HCC.
肝细胞癌（HCC）是全球最常见的恶性肿瘤之一，其发病机制尚不清楚。鞘磷脂磷酸二酯酶酸性样3A（SMPDL3A）影响细胞分化并参与免疫调节。然而，其在HCC中的分子生物学功能尚未阐明。
分析180例HCC患者的数据，以研究肝癌组织中SMPDL3A的表达与肝癌患者预后之间的关系。使用Crispr-Cas9双载体慢病毒敲除HCC细胞系中的SMPDL3A。通过CCK8测定、克隆形成实验、细胞周期测定、细胞划痕实验、TUNEL实验和流式细胞术确定SMPDL3A对细胞活力的影响。在BALB/c裸鼠中进行异种移植瘤实验，证实SMPDL3A促进肿瘤生长。通过质谱分析和免疫共沉淀对SMPDL3A相互作用蛋白进行初步探索。
本研究表明，HCC组织中SMPDL3A的表达与癌旁组织不同。此外，SMPDL3A高表达患者的总生存率和无瘤生存率显著低于SMPDL3A低表达患者。SMPDL3A表达与异常凝血酶原诱导蛋白水平、肝硬化、肿瘤直径、微血管侵犯和巴塞罗那临床肝癌分期密切相关。因此，SMPDL3A是影响HCC患者无瘤生存率和总生存率的独立危险因素。一项使用Crispr-Cas9基因组编辑技术的研究揭示了其对细胞增殖、凋亡和迁移的敲除作用。细胞计数试剂盒-8测定和克隆形成实验表明，sgSMPDL3A抑制肿瘤细胞增殖和迁移。流式细胞术和TUNEL测定表明，sgSMPDL3A促进肿瘤细胞凋亡。此外，sgSMPDL3A在裸鼠皮下肿瘤形成过程中抑制肿瘤生长。Ki67和PNCA的免疫组织化学也表明，sgSMPDL3A抑制荷瘤裸鼠皮下肿瘤增殖。进一步实验表明，SMPDL3A与原始同源物增强子（ERH）相互作用。
SMPDL3A高表达与HCC患者预后不良相关。敲除SMPDL3A可抑制HCC细胞的增殖和迁移并加速其凋亡。SMPDL3A与ERH相互作用以影响HCC的发生和发展。
