摘要
目的观察血管生成素-2 (Ang-2)在大鼠骨肉瘤建模中不同时期的表达及其对骨肉瘤细胞的调控作用。方法20只大鼠股骨下段注射MG63骨肉瘤细胞,成瘤后分为5、10、15、20d组,5只股骨下段注射等量生理盐水的大鼠为对照组。测定各组大鼠血清Ang-2含量,并对成瘤行免疫组化染色证实。构建敲减Ang-2基因表达的shRNA慢病毒载体,转染至培养的MG63骨肉瘤细胞中,观察对骨肉瘤细胞周期及凋亡的影响。结果实验组血清Ang-2在4个不同阶段均较对照组明显升高,其中10d组上升幅度最大(P值均<0.05);免疫组化染色发现,成瘤10d组Ang-2表达阳性细胞数最高,且与血清测定结果相符;抑制Ang-2表达使骨肉瘤细胞分裂阻滞在G1期,并增加细胞凋亡(P<0.05)。结论 Ang-2与骨肉瘤发生发展关系密切,可能成为骨肉瘤早期诊断及潜在治疗的一个新的参考指标。
Objective To observe the expression of angiogenin-2(Ang-2)at different stages in rat osteosarcoma modeling and its regulation effect on osteosarcoma cells.Methods MG63 osteosarcoma cells were injected into the lower femoral segment of 20 rats,and after tumor formation,the rats were divided into 5,10,15,and 20 days groups.Five rats were injected with the same amount of normal saline in the lower femoral segment as the control group.The serum Ang-2 content in each group was determined and confirmed by immunohistochemical staining.ShRNA lentivirus vector with ang-2 gene knockdown was constructed and transfected into cultured MG63 osteosarcoma cells to observe the effects on cell cycle and apoptosis of osteosarcoma cells.Results The serum Ang-2 level in the experimental group was significantly higher than that in the control group at four different stages,with the maximum increase in the 10-days group(all P<0.05).Immunohistochemical staining showed that the number of positive ang-2 cells was the highest in the 10-days tumorigenicity group,which was consistent with the serum test results.Inhibition of Ang-2 expression blocked cell division in the G1 phase and increased apoptosis(P<0.05).Conclusion Ang-2 is closely related to the occurrence and development of osteosarcoma,and may become a new reference index for early diagnosis and potential treatment of osteosarcoma.
作者
郭徽灵
胡世平
汤发强
吴宏
郑建章
GUO Huiling;HU Shiping;TANG Faqiang;WU Hong;ZHENG Jianzhang(The first Department of Orthopaedics,Fujian Provincial Hospital,Provincial Clinical Medical College of Fujian Medical University,Fuzhou,Fujian350001,China)
出处
《福建医药杂志》
CAS
2021年第2期135-138,F0004,共5页
Fujian Medical Journal
基金
2018年省卫生计生青年科研课题资助项目(2018-1-6)。
