骨肉瘤患者肿瘤组织中PLOD2基因表达水平及其对肿瘤细胞迁移/侵袭的影响  

Expression level of PLOD2 gene in tumor tissues of patients with osteosarcoma and its effects on tumor cell migration/invasion

作　　者：樊李瀛[1] 衡立松 段虹昊 程斌[1] 张纯[1] 姬乐[4] 臧全金[1] 吴昊[1] FAN Li-ying;HENG Li-song;DUAN Hong-hao(Department of Orthopaedics,the Second Affiliated Hospital of Xi'an Jiaotong University,Xi'an Shaanxi 710000,China;Department of Orthopaedics,Xi'an Red Cross Hospital,Xi'an Shaanxi 710000,China;Department of Hand Surgery,Xi'an Red Cross Hospital Hand Center,Xi'an Shaanxi 710000,China)

机构地区：[1]西安交通大学第二附属医院骨科,陕西西安710000 [2]西安市红会医院骨科,陕西西安710000 [3]西安市红会医院手外中心,陕西西安710000 [4]陕西省人民医院骨科,陕西西安710000

出　　处：《临床和实验医学杂志》2019年第24期2589-2593,共5页Journal of Clinical and Experimental Medicine

基　　金：国家自然科学基金(编号:81571209);陕西省重点研发计划一般项目(编号:2018SF-178)

摘　　要：目的 探讨骨肉瘤患者肿瘤组织中赖氨酸羟化酶2(PLOD2)基因表达水平及其对肿瘤细胞迁移/侵袭的影响。方法 选取西安交通大学第二附属医院骨科采集的95例新鲜骨肉瘤患者肿瘤组织及其癌旁正常组织标本,采用免疫组化染色和实时荧光定量PCR(RT-PCR)方法检测其表达水平,分析PLOD2表达与临床病理特征的关系。取对数生长期人骨肉瘤细胞系MG-63,随机分为小干扰RNA(si RNA)-PLOD2组、阴性对照组(NC组)以及空白组,si RNA-PLOD2组转染si RNA-PLOD2干扰序列,NC组转染空载体,空白组不予任何处理。采用RT-PCR方法检测各组PLOD2基因表达水平,观察抑制PLOD2基因表达对骨肉瘤细胞迁移、侵袭能力的影响。结果 免疫组化染色结果显示,骨肉瘤肿瘤组织中PLOD2阳性表达率(96. 84%)明显高于癌旁正常组织(7. 37%),差异有显著性(P <0. 05);骨肉瘤肿瘤组织中PLOD2 m RNA表达水平明显高于癌旁正常组织,差异有显著性(P <0. 05);PLOD2表达与TNM分期、软组织浸润、淋巴结远处转移密切相关(P <0. 05),与Ⅰ、Ⅱ期、不存在软组织浸润以及未发生淋巴结远处转移骨肉瘤患者比较,Ⅲ、Ⅳ期、存在软组织浸润以及发生淋巴结远处转移骨肉瘤患者肿瘤组织中PLOD2表达水平明显升高(P <0. 05),而与骨肉瘤患者性别、年龄、肿瘤直径、组织学类型无关(P> 0. 05);转染48 h,si RNA-PLOD2组PLOD2基因表达水平明显低于NC组、空白组(P <0. 05),NC组、空白组PLOD2基因表达水平无显著差异(P> 0. 05);Transwell侵袭实验以及划痕实验结果显示,si RNA-PLOD2组骨肉瘤细胞穿透基底膜数量、迁移细胞数量明显少于NC组、空白组(P<0. 05),NC组、空白组骨肉瘤细胞穿透基底膜数量、迁移细胞数量无显著差异(P> 0. 05)。结论 骨肉瘤患者肿瘤组织中PLOD2基因表达水平高于癌旁正常组织,其表达水平与骨肉瘤患者肿瘤TNM分期、软组织浸润、淋巴结远处转移有关,抑制PLOObjective To explore the expression level of procollage-lysine-2-oxoglutarate-5-dioxygenase 2(PLOD2)gene in tumor tissues of patients with osteosarcoma and its effects on tumor cell migration/invasion.Methods The specimens of tumor tissues and adjacent normal tissues of 95 patients with fresh osteosarcoma collected from department of orthopedics of the Second Affiliated Hospital of Xi'an Jiaotong University were selected.Immunohistochemical staining and real-time quantitative PCR(RT-PCR)were used to detect the expression level,and the relationship between expression and clinicopathological characteristics was analyzed.The human osteosarcoma cell line MG-63 in logarithmic growth phase were collected and randomly divided into small interfering RNA(siRNA)-PLOD2 group,NC group and blank group.siRNA-PLOD2 group was transfected with siRNA-PLOD2 interference sequence,and NC group was transfected with empty vector,and blank group was not treated,and the expression level of PLOD2 gene in each group was detected by RT-PCR,and the effects of inhibiting expression of PLOD2 gene on migration and invasion of osteosarcoma cells were observed.Results Immunohistochemical staining showed that the positive expression rate of PLOD2 in osteosarcoma tumor tissues was significantly higher than that in adjacent normal tissues(96.84%vs 7.37%)(P<0.05).The expression level of PLOD2 gene in osteosarcoma tumor tissues was significantly higher than that in adjacent normal tissues(P<0.05).PLOD2 expression was closely related to TNM stage,soft tissue infiltration and distant lymph node metastasis(P<0.05).Compared with patients with stage I and II osteosarcoma and osteosarcoma patients without soft tissue infiltration and without distant lymph node metastasis,the expression level of PLOD2 was significantly increased in the tumor tissues of patients with stage III and IV osteosarcoma and osteosarcoma patients with soft tissue infiltration and with distant lymph node metastasis(P<0.05),but the expression was not related to gender,age,tumor diamete

关 键 词：骨肉瘤 赖氨酸羟化酶 2 小干扰 RNA 细胞迁移 细胞侵袭 

分 类 号：R73[医药卫生—肿瘤]