摘要
目的:探究谷氨酰胺(Gln)对甲状腺癌细胞恶性生物学行为及SNX10/mTORC1通路蛋白表达的影响。方法:分别用含0、1、2、4 mmol/L Gln的培养基培养人未分化甲状腺癌细胞系C643,每个浓度设6个复孔。分组培养24、48、72 h后,MTT法检测细胞增殖活性。分组培养24 h后,采用Annexin V/PI双染法、划痕实验、Transwell小室实验、三磷酸腺苷(ATP)实验检测细胞凋亡率、迁移能力、侵袭能力、ATP生成能力,采用实时荧光定量PCR法检测SNX10、p-mTOR、mTORC1 mRNA的表达,Western blot法检测SNX10、p-mTOR、mTORC1、Akt、p-Akt蛋白的表达。结果:Gln干预后,C643细胞凋亡率下降,增殖活性、划痕愈合率、侵袭细胞数、ATP浓度均增加,SNX10、p-mTOR、mTORC1 mRNA相对表达量升高,SNX10、p-mTOR、mTORC1、Akt、p-Akt蛋白表达上调,且均呈剂量依赖性(P<0.05)。结论:Gln可能通过激活SNX10/mTORC1通路而促进甲状腺癌细胞的恶性生物学行为。
Aim:To explore the effect of glutamine(Gln)on the malignant biological behaviors and the expression of SNX10/mTORC1 pathway proteins of thyroid cancer cells.Methods:Human undifferentiated thyroid cancer cell line C643 was cultured and treated with 0,1,2,4 mmol/L Gln,respectively,each concentration had 6 complex holes.After 24,48,72 hours culture,MTT assay was used to detect cell proliferative activity.After 24 hours culture,Annexin V/PI staining method,scratch assay,Transwell assay,ATP assay were used to detect apoptosis,migration ability,invasion ability,and ATP-producing ability,RT-qPCR was used to detect the mRNA expressions of SNX10,p-mTOR,mTORC1,and the expressions of SNX10,p-mTOR,mTORC1,Akt,p-Akt proteins were detected by Western blot.Results:After Gln culture,the apoptosis rate of C643 cells decreased,the proliferative activity,scratch healing rate,number of invasive cells and the concentration of ATP increased,the mRNA relative expressions of SNX10,p-mTOR and mTORC1 increased,and the protein expressions of SNX10,p-mTOR,mTORC1,Akt and p-Akt were up-regulated in a dose-dependent manner(P<0.05).Conclusion:Gln may improve the malignant biological behaviors of thyroid cancer cells through activating SNX10/mTORC1 pathway.
作者
闫志华
程兵
陈伟娜
李祥周
YAN Zhihua;CHENG Bing;CHEN Weina;LI Xiangzhou(Department of Nuclear Medicine,the First Affiliated Hospital,Zhengzhou University,Zhengzhou 450052)
出处
《郑州大学学报（医学版）》
CAS
北大核心
2024年第4期544-548,共5页
Journal of Zhengzhou University(Medical Sciences)
