Abstract：
OBJECTIVE: In developed countries, endometrial cancer (EC) is the most prevalent gynecological cancer and its occurrence is associated with chronic inflammation. ATP5F1D is a subunit of ATP synthase (complex V), as well as the important component of mitochondrial electron transport chain (ETC). ETC play compelling roles in carcinogenesis. To date, little is known about the role of ATP5F1D in EC.
METHODS: ATP5F1D expression was identified in EC tissues and EC cell lines. We evaluated the influence of ATP5F1D on clinical features and prognosis based on TCGA database. The effects of ATP5F1D in EC malignant progression by applying loss-of-function assays in KLE and Ishikawa cell lines were detected by EdU, CCK-8, wound healing, Transwell, and flow cytometry assays. Additionally, electron microscope, LDH release, ELISA, mitochondrial ROS measurement, and Immunofluorescence were performed to demonstrate ATP5F1D can affect the pyroptosis of EC. To observe the anti-tumor effect on ATP5F1D silencing, we established an in vivo human endometrial tumor model using nude mice.
RESULTS: ATP5F1D expression was significantly upregulated in EC and was associated with favorable prognosis. ATP5F1D knockdown inhibited the proliferation, invasion, and migration of EC cells. Similarly, in nude mice, ATP5F1D knockdown suppressed the growth EC cells. Knocking down ATP5F1D lead to decrease the production of mitochondrial ROS (mtROS) and inhibited pyroptosis of EC cells.
CONCLUSIONS: Downregulation of ATP5F1D as a new therapeutic strategy that could mediate pyroptosis via suppressing mtROS/NLRP3/caspase-1/GSDMD pathway to inhibit EC progression.
METHODS: ATP5F1D expression was identified in EC tissues and EC cell lines. We evaluated the influence of ATP5F1D on clinical features and prognosis based on TCGA database. The effects of ATP5F1D in EC malignant progression by applying loss-of-function assays in KLE and Ishikawa cell lines were detected by EdU, CCK-8, wound healing, Transwell, and flow cytometry assays. Additionally, electron microscope, LDH release, ELISA, mitochondrial ROS measurement, and Immunofluorescence were performed to demonstrate ATP5F1D can affect the pyroptosis of EC. To observe the anti-tumor effect on ATP5F1D silencing, we established an in vivo human endometrial tumor model using nude mice.
RESULTS: ATP5F1D expression was significantly upregulated in EC and was associated with favorable prognosis. ATP5F1D knockdown inhibited the proliferation, invasion, and migration of EC cells. Similarly, in nude mice, ATP5F1D knockdown suppressed the growth EC cells. Knocking down ATP5F1D lead to decrease the production of mitochondrial ROS (mtROS) and inhibited pyroptosis of EC cells.
CONCLUSIONS: Downregulation of ATP5F1D as a new therapeutic strategy that could mediate pyroptosis via suppressing mtROS/NLRP3/caspase-1/GSDMD pathway to inhibit EC progression.
摘要：
目标：在发达国家，子宫内膜癌（EC）是最常见的妇科肿瘤，其发生与慢性炎症有关。ATP5F1D是ATP合酶（复合物V）的一个亚基，以及线粒体电子传递链(ETC)的重要构成部门。ETC在癌变中起着令人信服的作用。迄今为止,关于ATP5F1D在EC中的作用知之甚少。
方法：在EC组织和EC细胞系中鉴定了ATP5F1D的表达。我们基于TCGA数据库评估了ATP5F1D对临床特征和预后的影响。通过在KLE和Ishikawa细胞系中应用功能丧失测定法检测ATP5F1D在EC恶性进展中的作用，CCK-8，伤口愈合，Transwell,和流式细胞术测定。此外,电子显微镜,LDH释放,ELISA,线粒体ROS测量，免疫荧光显示ATP5F1D可影响EC的焦亡。观察ATP5F1D沉默的抗肿瘤作用,我们使用裸鼠建立了体内人子宫内膜肿瘤模型。
结果：ATP5F1D表达在EC中显著上调，并与良好的预后相关。ATP5F1D敲低抑制细胞增殖，入侵,和EC细胞的迁移。同样，在裸鼠中，ATP5F1D敲低抑制了EC细胞的生长。敲除ATP5F1D导致线粒体ROS（mtROS）的产生减少并抑制EC细胞的焦亡。
结论：下调ATP5F1D作为一种新的治疗策略，可以通过抑制mtROS/NLRP3/caspase-1/GSDMD通路来抑制EC进展，从而介导细胞凋亡。
方法：在EC组织和EC细胞系中鉴定了ATP5F1D的表达。我们基于TCGA数据库评估了ATP5F1D对临床特征和预后的影响。通过在KLE和Ishikawa细胞系中应用功能丧失测定法检测ATP5F1D在EC恶性进展中的作用，CCK-8，伤口愈合，Transwell,和流式细胞术测定。此外,电子显微镜,LDH释放,ELISA,线粒体ROS测量，免疫荧光显示ATP5F1D可影响EC的焦亡。观察ATP5F1D沉默的抗肿瘤作用,我们使用裸鼠建立了体内人子宫内膜肿瘤模型。
结果：ATP5F1D表达在EC中显著上调，并与良好的预后相关。ATP5F1D敲低抑制细胞增殖，入侵,和EC细胞的迁移。同样，在裸鼠中，ATP5F1D敲低抑制了EC细胞的生长。敲除ATP5F1D导致线粒体ROS（mtROS）的产生减少并抑制EC细胞的焦亡。
结论：下调ATP5F1D作为一种新的治疗策略，可以通过抑制mtROS/NLRP3/caspase-1/GSDMD通路来抑制EC进展，从而介导细胞凋亡。
