中国癌症杂志 ›› 2021, Vol. 31 ›› Issue (1): 20-26.doi: 10.19401/j.cnki.1007-3639.2021.01.003

• 论著 • 上一篇    下一篇

下调SLC6A3对肾透明细胞癌细胞SNU-349增殖、迁移的影响

王 佳 1 ,艾中中 2 ,周 菁 3   

  1. 1. 空军军医大学西京医院泌尿外科,陕西 西安 710000 ;
    2. 西安医学院第二附属医院口腔科,陕西 西安 710000 ;
    3. 陕西省肿瘤医院肿瘤内科,陕西 西安 710061
  • 出版日期:2021-01-30 发布日期:2021-02-20
  • 通信作者: 周 菁 E-mail: zhoujshanxi@163.com

Effects of SLC6A3 downregulation on proliferation and migration of SNU-349 cells in renal clear cell carcinoma

WANG Jia 1 , AI Zhongzhong 2 , ZHOU Jing 3   

  1. 1. Department of Urology Xijing Hospital, Air Force Medical University, Xi’an 710000, Shaanxi Province, China; 2. Department of Stomatology, the Second Affiliated Hospital of Xi’an Medical University, Xi’an 710000, Shaanxi Province, China; 3. Department of Oncology, Shaanxi Cancer Hospital, Xi’an 710061, Shaanxi Province, China
  • Published:2021-01-30 Online:2021-02-20
  • Contact: ZHOU Jing E-mail: zhoujshanxi@163.com

摘要: 背景与目的:溶质载体家族6成员3(solute carrier family 6 member 3,SLC6A3)在肾透明细胞癌患者中呈高表达,但SLC6A3对肾透明细胞癌细胞转移的影响及分子机制尚不明确。探讨SLC6A3对肾透明细胞癌细胞增殖、迁移的影响及潜在的分子机制。方法:采用免疫组织化学法检测2017年1月—2018年1月在陕西省肿瘤医院收集的56例肾透明细胞癌患者的癌组织及癌旁组织中SLC6A3的表达。采用实时荧光定量聚合酶链反应(real-time fluorescence quantitative polymerase chain reaction,RTFQ-PCR)和蛋白质印迹法(Western blot)检测SLC6A3在肾透明细胞癌细胞SNU-349中的表达。构建shNC和shSLC6A3质粒,转染至SNU-349细胞中,采用细胞计数试剂盒-8(cell counting kit-8,CCK-8)法、流式细胞术、transwell实验检测SLC6A3下调后细胞增殖、凋亡、侵袭能力的变化。采用Western blot分析SLC6A3下调对磷脂酰肌醇3激酶(phosphoinositide 3-kinase,PI3K)/蛋白激酶B(protein kinase B,Akt)信号转导通路的影响。结果:肾透明细胞癌组织及细胞中SLC6A3呈高表达(P<0.05)。下调SLC6A3后,细胞增殖、侵袭能力显著降低(P<0.001),凋亡率显著升高(P<0.05),细胞周期S期阻滞(P<0.05)。下调SLC6A3后细胞内PI3K及Akt磷酸化水平显著降低(P<0.05)。结论:SLC6A3在肾透明细胞癌组织中呈高表达,下调SLC6A3抑制细胞增殖、侵袭,促进细胞凋亡。SLC6A3可能通过调控PI3K/Akt信号通路影响SNU-349细胞的生长。

关键词: 肾透明细胞癌, 溶质载体家族6成员3, 增殖, 迁移

Abstract: Background and purpose: Solute carrier family 6 member 3 (SLC6A3) is highly expressed in patients with renal clear cell carcinoma. However, the effect of SLC6A3 on the metastasis of renal clear cell carcinoma and its molecular mechanism are still unclear. This study aimed to investigate the effect of SLC6A3 on proliferation and migration of renal clear cell carcinoma cells and its potential molecular mechanism. Methods: The expression of SLC6A3 in renal clear cell carcinoma and adjacent tissues collected in Shaanxi Cancer Hospital from January 2017 to January 2018 was detected by immunohistochemistry. The expression of SLC6A3 in renal clear cell carcinoma cell line SNU-349 was detected by real-time fluorescence quantitative polymerase chain reaction (RTFQ-PCR) and Western blot. ShNC and shSLC6A3 plasmids were constructed and transfected into SNU-349 cells. Changes in cell proliferation, apoptosis and invasion ability of SLC6A3 downregulated cells were detected by cell counting kit-8 (CCK-8) assay, flow cytometry and transwell assay. The effects of SLC6A3 downregulation on phosphoinositide 3-kinase (PI3K)/ protein kinase B (Akt) signaling pathway were analyzed by Western blot. Results: SLC6A3 was highly expressed in renal clear cell carcinoma tissues and cells (P<0.05). After SLC6A3 was downregulated, cell proliferation and invasion ability were significantly decreased (P<0.05), apoptotic rate was significantly increased (P<0.05), and cell cycle S-phase arrest was observed (P<0.05). The expression levels of PI3K and phosphorylation levels of Akt in cells were significantly reduced after SLC6A3 was downregulated (P<0.05). Conclusion: SLC6A3 is highly expressed in renal clear cell carcinoma tissues. Downregulation of SLC6A3 inhibits cell proliferation and invasion, and promotes cell apoptosis. SLC6A3 may affect the growth of SNU-349 cells by regulating the PI3K/Akt signaling pathway.

Key words: Renal clear cell carcinoma, Solute carrier family 6 member 3, Proliferation, Migration