Abstract

In this study, we explored the effects of treating human endometrial cancer cells with γ-synuclein-specific short hairpin RNA (shRNA) and elucidated the associated mechanisms in vitro and in vivo through the p38, extracellular signal-regulated kinase (ERK), and c-Jun N-terminal kinase (JNK) signaling pathways. Cell proliferation and migration were assessed using CCK8, Transwell, and scratch wound healing assays. Flow cytometry and laser scanning confocal microscopy were used to detect cell cycle changes. Relative levels of phosphorylated and non-phosphorylated (p) p38, ERK1/2 and JNK1/2/3 were determined in vitro and in vivo using simple western blotting assays. Cell proliferation in the experimental group decreased significantly and cells transfected with shRNA showed reduced migration rates (P < 0.05). p-p38, p-ERK1/2, and p-JNK1/2/3 levels were downregulated in the experimental group in vitro and in vivo. Tumor volumes and weights in the experimental group were significantly lower (P < 0.05). Tumor formation time in the negative control group was significantly shorter (P < 0.05). Flow cytometry showed that the number of cells in the G1 and mitotic phases increased and that in the S phase decreased after SNCG silencing (P < 0.05). Confocal microscopy showed that the percentage of cells in the mitotic phase increased after SNCG gene silencing (P < 0.05). We conclude that shRNA-mediated suppression of γ-synuclein decreased the proliferation, migration, and tumorigenicity of endometrial cancer cells via downregulation of p38, ERK, and JNK phosphorylation. High SNCG expression is closely related to the growth cycle of endometrial cancer cells.

中文翻译：

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shRNA介导的γ-突触核蛋白抑制抑制导致子宫内膜癌细胞p38 / ERK / JNK磷酸化和细胞周期阻滞下调

摘要

在这项研究中，我们探索了使用γ-突触核蛋白特异性短发夹RNA（shRNA）治疗人子宫内膜癌细胞的效果，并通过p38，细胞外信号调节激酶（ERK）和体外阐明了体内和体外的相关机制。 c-Jun N末端激酶（JNK）信号通路。使用CCK8，Transwell和刮擦伤口愈合试验评估细胞的增殖和迁移。流式细胞仪和激光扫描共聚焦显微镜用于检测细胞周期变化。磷酸化和非磷酸化（p）p38，ERK1 / 2和JNK1 / 2/3的相对水平是在体外和体内使用简单的蛋白质印迹法测定的。实验组细胞增殖明显下降，shRNA转染细胞迁移率降低（P<0.05）。在体外和体内实验组中，p -p38，p -ERK1 / 2和p -JNK1 / 2/3的水平下调。实验组的肿瘤体积和重量显着降低（P <0.05）。阴性对照组的肿瘤形成时间明显缩短（P <0.05）。流式细胞仪检测表明，SNCG沉默后，G1期和有丝分裂期的细胞数量增加，S期减少（P <0.05）。共聚焦显微镜显示SNCG基因沉默后有丝分裂期细胞的百分比增加（P<0.05）。我们得出的结论是，shRNA介导的γ-突触核蛋白抑制可通过下调p38，ERK和JNK磷酸化来降低子宫内膜癌细胞的增殖，迁移和致瘤性。SNCG的高表达与子宫内膜癌细胞的生长周期密切相关。