摘要
目的:研究溶质载体家族6成员9(solute carrier family 6 member 9,SLC6A9)表达对结直肠癌细胞增殖、迁移和5-氟尿嘧啶(5-fluorouracil,5-FU)药物敏感性的影响。方法:TCGA数据库分析、实时荧光定量PCR和Western blot分析检测SLC6A9在结肠癌组织、正常结肠细胞系(NCM460)和结直肠癌细胞系(SW620、HCT116、HT29、Lovo和SW480)中的表达。将SCL6A9过表达质粒及阴性对照(SLC6A9 OE、Vector)转染HT29细胞,将SCL6A9小干扰RNA及阴性对照(SLC6A9 siRNA1#、siRNA2#和Scramble)转染SW620细胞。划痕愈合实验和Transwell实验检测各组细胞的迁移、侵袭能力。Western blot和细胞免疫荧光检测EMT相关蛋白E-cadherin、Vimentin的表达水平。利用CCK-8法和构建裸鼠移植瘤模型检测SLC6A9过表达对结直肠癌细胞5-FU药物敏感性的影响。结果:与正常结肠组织和NCM460细胞相比,SLC6A9在结肠癌组织和结直肠癌细胞系中低表达(均P<0.05)。SLC6A9过表达引起E-cadherin蛋白表达增加,Vimentin蛋白水平降低,抑制结直肠癌细胞的迁移、侵袭(P<0.05)。SLC6A9低表达引起E-cadherin蛋白表达降低,Vimentin蛋白水平增加,促进结直肠癌细胞的迁移、侵袭能力(P<0.05)。SLC6A9过表达提高了5-FU的药物敏感性,并使肿瘤生长缓慢,质量减轻(P<0.05)。而SLC6A9低表达降低了5-FU的药物敏感性(P<0.05)。结论:SLC6A9过表达能够抑制结直肠癌细胞的迁移、侵袭和EMT进程,并增强5-FU对结直肠癌细胞的药物敏感性。
Objective:To investigate the expression of member 9 of the solute carrier family 6(SLC6A9)on the proliferation,migration,and sensitivity of colon cancer cells to 5-fluorouracil(5-FU).Methods:TCGA database analysis,real-time fluorescence quantitative PCR,and Western blot analysis were used to detect the expression of SLC6A9 in colon cancer tissues,normal colon cell lines(NCM460),and colorectal cancer cell lines(SW620,HCT116,HT29,Lovo,and SW480).Transfect HT29 cells with SCL6A9 overexpression plasmids and negative controls(SLC6A9 OE,Vector),and transfect SW620 cells with SCL6A9 small interfering RNA and negative controls(SLC6A9 siRNA1#,siRNA2#and Scramble).The scratch healing experiment and transwell experiment were used to detect the migration and invasion ability of cells in each group.Western blot and cellular immunofluorescence were used to detect the expression levels of EMT related proteins E-cadherin and Vimentin.Using the CCK-8 method and constructing a nude mouse tumor model to detect the therapeutic effect of overexpression of SLC6A9 on 5-FU in colorectal cancer cells.Results:Compared with normal colon tissues and NCM460 cells,SLC6A9 was low expressed in colon cancer tissues and colorectal cancer cell lines(both P<0.05).Overexpression of SLC6A9 leads to an increase in E-cadherin protein expression and a decrease in Vimentin protein levels,which inhibits the migration and invasion of colorectal cancer cells(P<0.05).Low expression of SLC6A9 leads to a decrease in E-cadherin protein expression and an increase in Vimentin protein levels,promoting the migration and invasion ability of colorectal cancer cells(P<0.05).Overexpression of SLC6A9 promoted the efficacy of 5-FU and resulted in slow tumor growth and reduced mass(P<0.05).The low expression of SLC6A9 decreased the sensitivity of 5-FU(P<0.05).Conclusion:Overexpression of SLC6A9 can inhibit the migration,invasion,and EMT progression of colorectal cancer cells,and enhance the sensitivity of 5-FU on colorectal cancer cells.
作者
张岩
田素礼
周勇旭
刘昶
ZHANG Yan;TIAN Suli;ZHOU Yongxu;LIU Chang(Department of General Surgery,the Fourth Affiliated Hospital of Harbin Medical University,Heilongjiang Harbin 150001,China)
出处
《现代肿瘤医学》
CAS
2024年第7期1236-1241,共6页
Journal of Modern Oncology
基金
黑龙江省自然科学基金联合引导项目(编号:LH2019H079)。
