乳腺癌(BCA)是女性中最常见的癌症之一。新的证据表明,膜联蛋白A-9(ANXA9)在某些癌症的发展中起着至关重要的作用。值得注意的是,ANXA9已被报道为胃癌和结直肠癌的新的预后生物标志物。然而,其在BCA中的表达和生物学功能尚未被研究。使用在线生物信息学工具,如TIMER,GEPIA,HPA,UALCAN,我们预测了ANXA9的表达及其与BCA患者临床病理特征的相关性。RT-qPCR和蛋白质印迹用于测量BCA患者组织和细胞中的ANXA9mRNA和ANXA9蛋白表达。通过透射电子显微镜鉴定BCA衍生的外来体。功能试验用于评估ANXA9在BCA细胞增殖中的生物学作用。迁移,入侵,和凋亡。使用肿瘤异种移植物体内模型来评估ANXA9在小鼠肿瘤生长中的作用。生物信息学和功能筛选分析显示ANXA9在BCA患者组织中高表达,ANXA9表达中位数比正常组织高1.5至2倍(p<0.05)。RT-qPCR证实BCA组织中的ANXA9表达比邻近正常组织高约1.5倍(p<0.001)。ANXA9在分歧亚型BCA中的表达也显示出差别,发现ANXA9在腔内BCA中相对于正常组织或其他组织学亚型大多显著上调(p<0.001)。此外,ANXA9在不同种族中表达升高,年龄,临床分期,淋巴结转移状态,和绝经状态组相对于正常组(p<0.001)。此外,发现ANXA9由BCA组织来源的外泌体分泌,其表达在外泌体处理的BCA细胞中上调1至7倍(p<0.001),而其在MCF10A细胞中的表达通过外泌体处理没有显着改变(p>0.05)。ANXA9沉默诱导BCA细胞集落数显着减少约30%(p<0.01)。迁移和入侵的BCA细胞的数量也减少了约65%和68%,分别,沉默ANXA9后(p<0.01)。在异种移植模型中,相对于LV-NC组,LV-sh-ANXA9组的肿瘤大小显着减小(近一半)(p<0.01),表明ANXA9沉默在体外和体内抑制了BCA进展中的肿瘤进展。总之,外泌体衍生的ANXA9作为促进增殖的癌基因发挥作用,迁移,和BCA细胞的侵袭性,并在BCA发育中增强肿瘤的生长,这可能为BCA患者提供新的预后和治疗生物标志物。
Breast cancer (BCA) is one of the most prevalent cancers among women. Emerging evidence has revealed that Annexin A-9 (
ANXA9) plays a crucial function in the development of some cancers. Notably,
ANXA9 has been reported to be a new prognostic biomarker for gastric and colorectal cancers. However, its expression and biological function in BCA have not yet been investigated. Using online bioinformatics tools such as TIMER, GEPIA, HPA, and UALCAN, we predicted ANXA9 expression and its correlation with the clinicopathological characteristics of BCA patients. RT-qPCR and western blot were utilized to measure
ANXA9 mRNA and
ANXA9 protein expression in BCA patient tissues and cells. BCA-derived exosomes were identified by transmission electron microscopy. Functional assays were employed to evaluate the biological role of ANXA9 in BCA cell proliferation, migration, invasion, and apoptosis. A tumor xenograft in vivo model was utilized to assess the role of ANXA9 in tumor growth in mice. Bioinformatics and functional screening analysis revealed that ANXA9 was highly expressed in BCA patient tissues, with median
ANXA9 expression 1.5- to 2-fold higher than in normal tissues (p < 0.05). RT-qPCR confirmed that ANXA9 expression in BCA tissues was around 1.5-fold higher than the adjacent normal tissues (p < 0.001).
ANXA9 expression in different subtypes of BCA also showed a difference, and
ANXA9 was found to be mostly significantly upregulated in luminal BCA relative to normal tissues or other histological subtypes (p < 0.001). Moreover, ANXA9 expression was elevated in different races, ages, clinical stages, node metastasis status, and menopause status groups relative to the normal group (p < 0.001). Furthermore, ANXA9 was found to be secreted by BCA tissue-derived exosomes and its expression was upregulated 1- to 7-fold in BCA cells treated with exosomes (p < 0.001), while its expression in MCF10A cells was not significantly altered by treatment with exosomes (p > 0.05). ANXA9 silencing induced a significant decrease of around 30% in the colony number of BCA cells (p < 0.01). The number of migrated and invaded BCA cells also decreased by around 65 and 68%, respectively, after silencing ANXA9 (p < 0.01). Tumor size was significantly reduced (nearly half) in the LV-sh-ANXA9 group relative to the LV-NC group in the xenograft model (p < 0.01), suggesting that ANXA9 silencing repressed tumor progression in BCA progression in vitro and in vivo. In conclusion, exosome-derived ANXA9 functions as an oncogene that facilitates the proliferation, migration, and invasiveness of BCA cells and enhances tumor growth in BCA development, which may provide a new prognostic and therapeutic biomarker for BCA patients.