Luo Jie, Ding Li, Pan Shirui, Luo Jing, Zhao Haiqiu, Yin Jiaxiu, Su Rong, Zhang Jiamin, Liu Lin
Department of Hematology of the First Affiliated Hospital of Chongqing Medical University, Chongqing, China.
Department of Hematology, The Affiliated Hospital of Southwest Medical University, Luzhou, China.
Front Pharmacol. 2024 Oct 23;15:1390456. doi: 10.3389/fphar.2024.1390456. eCollection 2024.
As a malignant hematological disease, the incidence of acute myeloid leukemia (AML) has exhibited an upward trend in recent years. Nevertheless, certain limitations persist in the treatment of AML. Sperm-associated antigen 6 (SPAG6) has been implicated in the onset and progression of various human cancers, with its expression levels significantly elevated in AML. Consequently, we undertook a series of experiments to investigate the role and underlying mechanisms of SPAG6 in AML cell lines.
In the experiments of this study, DEPs and GO and KEGG enrichment analysis subsequent to SPAG6 down-regulation were detected by TMT. CCK8 was employed to determine cell viability. The levels of apoptosis and ROS were measured by flow cytometry. In the experiments, a xenografted tumor model was constructed, and the expression of SPAG6 and GSTP1 in tumor tissues was detected by IHC.
Ultimately, our findings indicated that over-expression of SPAG6 promoted cell growth and decreased reactive oxygen species (ROS) and malondialdehyde levels. Furthermore, SPAG6 knockdown was found to diminish mitochondrial membrane potential and facilitate cell apoptosis. , SPAG6 could also promote tumor growth, suggesting that SPAG6 may serve as a pro-tumor factor. In addition, daunorubicin (DNR) may cause oxidative stress and initiate apoptosis, resulting in oxidative damage to AML cells. However, the overexpression of SPAG6 may attenuate the efficacy of DNR. This was due to SPAG6 promoted GSTP1 expression, thereby reducing ROS levels. Simultaneously, the elevation of GSTP1 and JNK complex may reduce the expression of p-JNK and inhibit the activation of JNK pathway, which might inhibit cell apoptosis.
In conclusion, our experiments suggested that upregulated SPAG6 might mitigate the pro-apoptotic effects of DNR through ROS/JNK MAPK axis in a GSTP1-dependent manner.
作为一种恶性血液病，近年来急性髓系白血病（AML）的发病率呈上升趋势。然而，AML的治疗仍存在一定局限性。精子相关抗原6（SPAG6）与多种人类癌症的发生和发展有关，其在AML中的表达水平显著升高。因此，我们进行了一系列实验来研究SPAG6在AML细胞系中的作用及潜在机制。
在本研究实验中，通过TMT检测SPAG6下调后的差异表达蛋白（DEPs）以及基因本体（GO）和京都基因与基因组百科全书（KEGG）富集分析。采用CCK8法测定细胞活力。通过流式细胞术检测凋亡和活性氧（ROS）水平。在实验中，构建异种移植肿瘤模型，通过免疫组织化学（IHC）检测肿瘤组织中SPAG6和谷胱甘肽S转移酶P1（GSTP1）的表达。
最终，我们的研究结果表明，SPAG6的过表达促进细胞生长，并降低活性氧（ROS）和丙二醛水平。此外，发现敲低SPAG6可降低线粒体膜电位并促进细胞凋亡。此外，SPAG6还可促进肿瘤生长，表明SPAG6可能是一种促肿瘤因子。此外，柔红霉素（DNR）可能会引起氧化应激并引发凋亡，导致AML细胞发生氧化损伤。然而，SPAG6的过表达可能会减弱DNR的疗效。这是因为SPAG6促进了GSTP1的表达，从而降低了ROS水平。同时，GSTP1和c-Jun氨基末端激酶（JNK）复合物的升高可能会降低磷酸化JNK（p-JNK）的表达并抑制JNK途径的激活，这可能会抑制细胞凋亡。
总之，我们的实验表明，上调的SPAG6可能通过ROS/JNK丝裂原活化蛋白激酶（MAPK）轴以GSTP1依赖的方式减轻DNR的促凋亡作用。
