Abstract：
Vesicle-associated membrane protein 8 (VAMP8), a soluble n-ethylmaleimide-sensitive factor receptor protein, acts as an oncogenic gene in the progression of several malignancies. Nevertheless, the roles and mechanisms of VAMP8 in colorectal cancer (CRC) progression remain unknown. The expression and prognostic significance of VAMP8 in CRC samples were analyzed through bioinformatics analyses. Cell proliferation was detected using CCK-8 and EdU incorporation assays and apoptosis was evaluated via flow cytometry. Western blot analysis was conducted to examine the protein expression. Ferroptosis was evaluated by measurement of iron metabolism, lipid peroxidation, and glutathione (GSH) content. VAMP8 was increased in CRC samples relative to normal samples on the basis of GEPIA and HPA databases. CRC patients with high level of VAMP8 had a worse overall survival. VAMP8 depletion led to a suppression of proliferation and promotion of apoptosis in CRC cells. Additionally, VAMP8 knockdown suppressed beclin1 expression and LC3-II/LC3-I ratio, elevated p62 expression, increased Fe2+, labile iron pool, lipid reactive oxygen species, and malondialdehyde levels, and repressed GSH content and glutathione peroxidase activity. Moreover, VAMP8 knockdown inhibited the activation of janus kinase (JAK)/signal transducer and activator of transcription 3 (STAT3) pathway in CRC cells. Mechanistically, activation of the JAK/STAT3 pathway by JAK1 or JAK2 overexpression attenuated VAMP8 silencing-mediated anti-proliferative, pro-apoptotic, anti-autophagic, and pro-ferroptotic effects on CRC cells. In conclusion, VAMP8 knockdown affects the proliferation, apoptosis, autophagy, and ferroptosis by the JAK/STAT3 pathway in CRC cells.
摘要：
囊泡相关膜蛋白8（VAMP8），可溶性正乙基马来酰亚胺敏感因子受体蛋白，在几种恶性肿瘤的进展中充当致癌基因。然而，VAMP8在结直肠癌(CRC)进展中的作用和机制尚不清楚.通过生物信息学分析分析VAMP8在CRC样本中的表达和预后意义。使用CCK-8和EdU掺入测定检测细胞增殖,并通过流式细胞术评估细胞凋亡。进行蛋白质印迹分析以检查蛋白质表达。通过测量铁代谢来评估铁凋亡，脂质过氧化，和谷胱甘肽(GSH)含量。基于GEPIA和HPA数据库,CRC样品中的VAMP8相对于正常样品增加。高VAMP8水平的CRC患者的总体生存率较差。VAMP8耗竭导致CRC细胞增殖抑制和凋亡促进。此外,VAMP8敲低抑制beclin1表达和LC3-II/LC3-I比率，p62表达升高，增加Fe2+，不稳定的铁池，脂质活性氧，和丙二醛水平，并抑制了GSH含量和谷胱甘肽过氧化物酶活性。此外，VAMP8敲低抑制CRC细胞中Janus激酶（JAK）/信号转导和转录激活因子3（STAT3）通路的激活。机械上,JAK1或JAK2过表达对JAK/STAT3通路的激活减弱了VAMP8沉默介导的抗增殖，促凋亡，抗自噬,和对CRC细胞的促铁蛋白作用。总之,VAMP8敲低影响增殖，凋亡，自噬,和通过JAK/STAT3途径在CRC细胞中的铁凋亡。
