Liu Aifeng, Huang Weiguo, Zeng Guqing, Ma Xiaohua, Zhou Xiao, Wang Yafei, Ouyang Chenjie, Cheng Ailan
Cancer Research Institute University of South China, Hengyang, Hunan 421001, P.R. China.
School of Nursing, University of South China, Hengyang, Hunan 421001, P.R. China.
Mol Med Rep. 2014 Dec;10(6):3059-67. doi: 10.3892/mmr.2014.2656. Epub 2014 Oct 15.
Nasopharyngeal carcinoma (NPC) has a highly increased incidence rate (20/100,000) in Southern regions of China, while being rare in the rest of the world. NPC is a malignant type of cancer due to its high occurrence rate of metastasis; however, biomarkers for effective diagnosis and treatment are yet to be identified. Annexin A1 is a glucocorticoid‑regulated member of a large superfamily of calcium and phospholipid‑binding proteins and has been shown to have important roles in tumor development and progression, and was demonstrated to be a prognostic biomarker for head and neck cancer types. A previous study by our group showed that Annexin A1 was decreased in NPC tissue as compared with normal adjacent tissue. To investigate whether Annexin A1 is a potential biomarker for NPC, the present study assessed the effect of the Annexin A1 on the biological behavior (i.e., invasion and metastasis) of the highly metastatic NPC cell line 5‑8F and the non‑metastatic NPC cell line 6‑10B. The expression levels of Annexin A1 in the above two cell lines were determined by western blot analysis. Next, the recombinant plasmid pEGFP‑C1‑Annexin A1 and the small interfering (si)RNA plasmid pRNAT‑U6.1‑Annexin A1 were used and stably transfected into 5‑8F and 6‑10B cells, respectively. These established recombinant cell lines were then used to study the up- and downregulation of Annexin A1, respectively. The correlation of Annexin A1 expression levels with the biological behavior of NPC cell lines was analyzed using a cell proliferation assay, flow cytometry, soft agar colony formation assay, as well as Transwell invasion and migration assays. The results demonstrated that upregulation of Annexin A1 suppressed the proliferation, invasion and migration of NPC cells, while downregulation of Annexin A1 promoted the proliferation, invasion and migration of NPC cells. These findings suggested that Annexin A1 may be a potential biomarker for the development and prognosis of NPC, and its dysregulation may have an important role in its underlying pathogenesis.
鼻咽癌(NPC)在中国南方地区的发病率极高(20/100,000),而在世界其他地区则较为罕见。由于其转移发生率高,鼻咽癌是一种恶性癌症类型;然而,尚未确定有效的诊断和治疗生物标志物。膜联蛋白A1是钙和磷脂结合蛋白的一个大型超家族中受糖皮质激素调节的成员,已被证明在肿瘤发展和进展中起重要作用,并被证明是头颈部癌症类型的预后生物标志物。我们小组之前的一项研究表明,与相邻正常组织相比,膜联蛋白A1在鼻咽癌组织中表达降低。为了研究膜联蛋白A1是否是鼻咽癌的潜在生物标志物,本研究评估了膜联蛋白A1对高转移性鼻咽癌细胞系5-8F和非转移性鼻咽癌细胞系6-10B生物学行为(即侵袭和转移)的影响。通过蛋白质免疫印迹分析确定上述两种细胞系中膜联蛋白A1的表达水平。接下来,分别将重组质粒pEGFP-C1-膜联蛋白A1和小干扰(si)RNA质粒pRNAT-U6.1-膜联蛋白A1用于稳定转染5-8F和6-10B细胞。然后使用这些建立的重组细胞系分别研究膜联蛋白A1的上调和下调。使用细胞增殖试验、流式细胞术、软琼脂集落形成试验以及Transwell侵袭和迁移试验分析膜联蛋白A1表达水平与鼻咽癌细胞系生物学行为的相关性。结果表明,膜联蛋白A1的上调抑制了鼻咽癌细胞的增殖、侵袭和迁移,而膜联蛋白A1的下调促进了鼻咽癌细胞的增殖、侵袭和迁移。这些发现表明,膜联蛋白A1可能是鼻咽癌发生发展和预后的潜在生物标志物,其失调可能在其潜在发病机制中起重要作用。