摘要
目的:探讨PD-1分子在急性T淋巴细胞白血病(T-ALL)患者来源的肿瘤细胞(T-ALL细胞)中的表达及其临床意义。方法:选用2015年12月江苏省中医院血液科提供的1例T-ALL细胞、4例健康志愿者提供的PBMC和博生吉医药科技(苏州)有限公司提供的人293T/PD-1细胞,将T-ALL细胞经尾静脉注射到B-NDG小鼠构建T-ALL细胞异种移植瘤模型,用流式细胞术检测移植瘤小鼠脾中获得的细胞是否主要是由T-ALL细胞组成。用流式细胞术检测T-ALL细胞中PD-1蛋白的表达,用RT-PCR进一步验证T-ALL细胞中PD-1 m RNA表达水平。对T-ALL细胞中PD-1基因进行SNP测序,以检测PD-1基因DNA序列是否发生改变。在体外使用PD-1抑制剂研究其对T-ALL细胞增殖、凋亡以及相关因子mRNA表达水平的影响。结果:成功构建小鼠TALL细胞异种移植瘤模型,用流式细胞术确认了该例疾病是T-ALL。T-ALL细胞中PD-1 mRNA和蛋白均高表达(均P<0.01)。PD-1基因的第5个外显子的一个碱基由胞嘧啶突变成胸腺嘧啶。在体外PD-1抑制剂对T-ALL细胞的增殖和凋亡均无明显影响;PD-1抑制剂上调抑癌蛋白IGFBP3 mRNA表达,降低促癌蛋白SULT1A3 mRNA表达(均P<0.01)。结论:PD-1在T-ALL细胞中高表达,PD-1可作为临床上T-ALL诊断及治疗的靶点。
Objective:To investigate the expression and clinical significance of PD-1 molecule in tumor cells(T-ALL cells)derived from the patient with T-cell acute lymphoblastic leukemia(T-ALL).Methods:T-ALL cells from one patient and PBMCs from four healthy volunteers provided by the Department of Hematology in Jiangsu Provincial Hospital of Traditional Chinese Medicine in December 2015,and human 293 T/PD-1 cells provided by Persongen Bio Therapeutics(Suzhou)Co.,Ltd.were used for this study.The mouse T-ALL xenograft model was constructed by injecting T-ALL cells into tail vein of B-NDG mice,and flow cytometry was used to verify whether the cells obtained from the spleen of transplanted mice were mainly consisted of T-ALL cells.Flow cytometry was used to study the protein expression of PD-1 in T-ALL cells,and RT-PCR was applied to further verify the m RNA expression of PD-1 in T-ALL cells.The PD-1 gene in T-ALL cells was sequenced by SNP genotyping to detect whether the DNA sequence of PD-1 gene changed.PD-1 inhibitor was used in vitro to study their effects on proliferation,apoptosis,and the m RNA expression levels of related factors in T-ALL cells.Results:The mouse T-ALL xenograft model was successfully constructed and verified by flow cytometry as T-ALL.PD-1 was highly expressed at both m RNA and protein levels in T-ALL cells(all P<0.01).A C-to-T mutation was detected in the fifth exon of the PD-1 gene.PD-1 inhibitor had no significant effect on proliferation and apoptosis of T-ALL cells in vitro;PD-1 inhibitor up-regulated the mRNA expression of tumor-suppressor protein IGFBP3 and decreased the mRNA expression of oncoprotein SULT1 A3(all P<0.01).Conclusion:PD-1 is highly expressed in T-ALL cells,and PD-1 could be used as a target for clinical diagnosis and treatment for T-ALL.
作者
温春媚
李自宣
王禹
朱学军
孟会敏
鞠杰
张亭亭
张秀艳
袁磊
安钢力
杨林
WEN Chunmei;LI Zixuan;WANG Yu;ZHU Xuejun;MENG Huimin;JU Jie;ZHANG Tingting;ZHANG Xiuyan;YUAN Lei;AN Gangli;YANG Lin(Cyrus Tang Hematology Center,Soochow University,Suzhou 215123,Jiangsu,China;Collaborative Innovation Center of Hematology,Soochow University,Suzhou 215123,Jiangsu,China;State Key Laboratory of Radiation Medicine and Protection,Soochow University,Suzhou 215123,Jiangsu,China;Persongen Bio Therapeutics (Suzhou) Co.,Ltd,Suzhou 215123,Jiangsu,China;Department of Hematology,Central Laboratory,the Affiliated Hospital of Nanjing University of Chinese Medicine,Jiangsu Provincial Hospital of Traditional Chinese Medicine,Nanjing 210023,Jiangsu,China;Department of Hematology,the Third Hospital of Peking University,Beijing 100191,China)
出处
《中国肿瘤生物治疗杂志》
CAS
CSCD
北大核心
2019年第7期768-775,共8页
Chinese Journal of Cancer Biotherapy
基金
国家自然科学基金资助项目(No.81872431,No.31471283)
国家重点研发计划(No.2016YFC1303403)
协同创新重大项目(No.XYXT2015304)
江苏省“六大人才高峰工程”(No.SWYY-CXTD-010)
江苏省科技发展计划(No.BE2016809)
南京市科技发展计划(No.201503011,No.18030801126)~~
