Department of Thoracic Surgery, China-Japan Friendship Hospital, Beijing, China.
Department of Thoracic Surgery, National Cancer Center/National Clinical Research Center for Cancer/Cancer Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China.
Cancer Med. 2023 Feb;12(3):3299-3312. doi: 10.1002/cam4.5079. Epub 2022 Jul 27.
Emerging evidence indicates that myristoylated alanine-rich C kinase substrate like 1 (MARCKSL1) is involved in the progression of esophageal squamous cell carcinoma (ESCC). However, the underpinning mechanism is unclear. Here, we investigated the mechanisms involving MARCKSL1 in ESCC progression.
CCK8, Transwell and wound-healing assays were employed to test the effect of MARCKSL1 on proliferation, invasion and migration in vitro. Next, transcriptome profiling was conducted through RNA sequencing to reveal the underlying mechanism of MARCKSL1 in ESCC progression, which was subsequently verified by western blot and qPCR analysis. Moreover, immunofluorescence and gelatin degradation assays were performed to reveal the ability of MARCKSL1 to mediate invadopodia formation and extracellular matrix (ECM) degradation. Finally, the correlation between MARCKSL1 and the clinicopathological features of ESCC patients was assessed based on TCGA database analysis and immunohistochemistry staining of tissue microarrays.
Knockdown of MARCKSL1 markedly attenuated the cell motility capacity of ESCC cells in vitro, while MARCKSL1 overexpression had the opposite effect. Transcriptomic analysis showed that MARCKSL1 mediated the mobility and migration of ESCC cells. In addition, overexpression of MARCKSL1 increased the colocalization of F-actin and cortactin at the frontier edge of migrating cells and ECM degradation. Furthermore, in ESCC patients, the mRNA level of MARCKSL1 in esophageal carcinomas (n = 182) was found to be notably higher than that in adjacent esophageal epithelia (n = 286) and the expression levels of MARCKSL1 in the tumor tissues (n = 811) were significantly increased compared to those in noncancerous esophageal tissues (n = 442) with a large sample size. Higher expression of MARCKSL1 was positively correlated with lymph node metastasis and associated with worse survival rates of patients with ESCC.
MARCKSL1 promotes cell migration and invasion by interacting with F-actin and cortactin to regulate invadopodia formation and ECM degeneration. High MARCKSL1 expression is positively correlated with poor prognosis in ESCC patients with lymph node metastasis.
越来越多的证据表明，豆蔻酰化丙氨酸丰富的蛋白激酶 C 底物样 1（MARCKSL1）参与了食管鳞状细胞癌（ESCC）的进展。然而，其潜在机制尚不清楚。在这里，我们研究了 MARCKSL1 在 ESCC 进展中的作用机制。
采用 CCK8、Transwell 和划痕愈合实验检测 MARCKSL1 对 ESCC 细胞体外增殖、侵袭和迁移的影响。接下来，通过 RNA 测序进行转录组谱分析，揭示 MARCKSL1 在 ESCC 进展中的潜在机制，随后通过 Western blot 和 qPCR 分析进行验证。此外，进行免疫荧光和明胶降解实验以揭示 MARCKSL1 介导侵袭小体形成和细胞外基质（ECM）降解的能力。最后，基于 TCGA 数据库分析和组织微阵列的免疫组织化学染色，评估 MARCKSL1 与 ESCC 患者临床病理特征的相关性。
MARCKSL1 敲低显著抑制 ESCC 细胞的体外运动能力，而 MARCKSL1 过表达则产生相反的效果。转录组分析表明 MARCKSL1 介导 ESCC 细胞的迁移和迁移能力。此外，MARCKSL1 的过表达增加了迁移细胞前沿边缘 F-肌动蛋白和桩蛋白的共定位和 ECM 的降解。此外，在 ESCC 患者中，182 例食管癌（n=182）中 MARCKSL1 的 mRNA 水平明显高于 286 例相邻食管上皮（n=286），811 例肿瘤组织（n=811）中 MARCKSL1 的表达水平明显高于 442 例非癌性食管组织（n=442）。MARCKSL1 表达水平较高与淋巴结转移呈正相关，并与 ESCC 患者的生存率降低相关。
MARCKSL1 通过与 F-肌动蛋白和桩蛋白相互作用，促进细胞迁移和侵袭，调节侵袭小体的形成和 ECM 的降解。MARCKSL1 高表达与淋巴结转移的 ESCC 患者预后不良呈正相关。
