谷氨酰胺转运载体SLC1A5对食管鳞癌细胞恶性生物学的影响及其机制  

The Impact and Mechanisms of Glutamine Transporter SLC1A5on the Malignant Biology of Esophageal Cancer Cells

作　　者：赵华山[1] 张旭东 陈乐乐 金星 周克旭 杜越 高社干[2] 张秀森 ZHAO Huashan;ZHANG Xudong;CHEN Lele;JIN Xing;ZHOU Kexu;DU Yue;GAO Shegan;ZHANG Xiusen(The First Affiliated Hospital(College of Clinical Medicine)of Henan University of Science and Technology,Luoyang,China,471003;State Key Laboratory of Esophageal Cancer Prevention&Treatment,Henan Key Laboratory of Microbiome and Esophageal Cancer Prevention and Treatment,Henan Key Laboratory of Cancer Epigenetics,Cancer Hospital,The First Affiliated Hospital(College of Clinical Medicine)of Henan University of Science and Technology,Luoyang,China,471003)

机构地区：[1]河南科技大学第一附属医院,全科医学科,洛阳471003 [2]河南科技大学临床医学院,河南科技大学第一附属医院,肿瘤医院,省部共建食管癌防治国家重点实验室,河南省微生态与食管癌防治重点实验室,河南省肿瘤表观遗传重点实验室,洛阳471003

出　　处：《食管疾病》2024年第1期8-11,23,共5页Journal of Esophageal Diseases

基　　金：河南省医学科技攻关计划省部共建项目(SB201902022);河南省中原英才计划(育才系列);河南省重点研发与推广专项(科技攻关)项目(222102310339);洛阳市科技发展重点项目(2202005A);河南省医学科技攻关计划联合共建项目(LHGJ20200571)。

摘　　要：目的探讨谷氨酰胺转运载体SLC1A5在食管鳞癌细胞系中的表达、对增殖和侵袭的影响及其作用机制。方法采用实时定量PCR及免疫印迹法检测人正常食管上皮细胞系(SHEE)和食管鳞癌细胞系(KYSE30、KYSE70、KYSE140、TE1)中SLC1A5的表达水平。在SLC1A5高表达的KYSE70细胞中采用CRISPR/Cas9方法敲除SLC1A5,采用CCK8测定和划痕实验检测对照、敲除SLC1A5和使用SLC1A5抑制剂V-9302后KYSE70细胞的增殖和迁移能力。采用GSH/GSSG试剂盒检测对照、敲除SLC1A5和使用SLC1A5抑制剂V-9302后KYSE70细胞的谷氨酰胺代谢情况。结果SLC1A5在食管癌细胞系KYSE70中高表达,与正常食管上皮中的表达有统计学差异(P<0.05)。SLC1A5敲除或被抑制的KYSE70细胞的增殖能力低于未处理细胞(P<0.05),处理组划痕的迁移面积较未处理组明显减少(P<0.05),处理组的GSH/GSSG比率较未处理组的显著降低(P<0.05)。结论SLC1A5在ESCC细胞系中表达增高,抑制SLC1A5可降低ESCC细胞的增殖迁移能力与谷氨酰胺代谢,SLC1A5可能是食管癌治疗的潜在靶点。Objective To investigate the expression of glutamine transporter SLC1A5 in esophageal squamous cell carcinoma cell lines and its impact on proliferation and invasion,as well as its underlying mechanisms.Methods Real-time quantitative PCR and immunoblotting were employed to measure the expression levels of SLC1A5 in human normal esophageal epithelial cell line(SHEE)and esophageal squamous cell carcinoma cell lines(KYSE30,KYSE70,KYSE140,TE1).The CRISPR/Cas9 system was used to knock out SLC1A5 in KYSE70 cells,which exhibited high expression level of SLC1A5.The proliferation and migration ability of esophageal cancer KYSE70 cells were measured using CCK8 assay and scratch assay between control,knockout of SLC1A5,and use of SLC1A5 inhibitor V-9302 groups.Glutamine metabolism in KYSE70 cells was examined by GSH/GSSG assay following the same conditions.Results SLC1A5 was found to be highly expressed in the esophageal squamous cell carcinoma cell line KYSE70,showing significant differences compared to its expression in normal esophageal epithelial cells(P<0.05).Upon knockout of SLC1A5 and treatment with the SLC1A5 inhibitor V-9302,the proliferation capacity of KYSE70 tumor cells was significantly reduced compared to the untreated cell group(P<0.05).The migration area of the scratch assay was significantly decreased in the treated groups compared to the untreated group(P<0.05).The GSH/GSSG ratio assay demonstrated that the knockout of SLC1A5 and treatment with the SLC1A5 inhibitor V-9302 led to a significant decrease in the GSH/GSSG ratio compared to the untreated group(P<0.05).Conclusion SLC1A5 is highly expressed in esophageal squamous cell carcinoma cell lines.Inhibiting SLC1A5 can effectively reduce the proliferation and migration abilities of esophageal cancer cells,as well as decrease glutamine metabolism.These findings provide new insights for the treatment of esophageal cancer.

关 键 词：SLC1A5 食管癌 增殖 凋亡 谷氨酰胺代谢 

分 类 号：R735.1[医药卫生—肿瘤]