丝氨酸/苏氨酸激酶31对骨肉瘤恶性生物学行为的影响  

The influence of STK31 on osteosarcoma′s malignant biological behavior

作　　者：莫坚[1] 朱江龙[1] 丰哲[1] 李书振[1] 韩杰[1] 苏波[1] 冯思坛[1] Mo Jian Zhu Jianglong Feng Zhe Li Shuzhen Han Jie Su Bo Feng Sitan(Department of Bone and Joint Surgery ,Ruikang Hospital Affiliated to Guangxi University of Traditional Chinese Medicine, Nanning , Guangxi 530011, China)

机构地区：[1]广西中医药大学附属瑞康医院骨关节科,南宁530011

出　　处：《重庆医学》2017年第23期3195-3197,3200,共4页Chongqing medicine

基　　金：国家自然科学基金地区科学基金项目(81560779)

摘　　要：目的观察丝氨酸/苏氨酸激酶31(STK31)在骨肉瘤中的表达及其对骨肉瘤恶性生物学行为的影响。方法收取15例骨肉瘤组织标本及瘤旁正常组织标本,利用免疫组织化学技术、实时定量PCR技术和Western blot技术检测肿瘤组织及瘤旁正常组织中STK31的表达;构建STK31敲除质粒pGenesil-STK31-shRNA,以骨肉瘤细胞系MG63细胞为靶细胞转染pGenesil-STK31-shRNA或对照质粒pGenesil-1,通过CCK8细胞活性实验,检测STK31对MG63细胞增殖能力的影响;Tanswell实验观察STK31对骨肉瘤细胞迁移能力的影响。结果免疫组织化学显示肿瘤组织中STK31表达明显高于瘤旁正常组织;实时定量PCR检测骨肉瘤STK31mRNA的表达量显著高于瘤旁正常组织[(3.65±0.83)vs.(1.05±0.14),P<0.05];Western blot显示肿瘤组织中STK31蛋白表达显著高于瘤旁正常组织(P<0.05);CCK8细胞活性实验显示敲除STK31后MG63细胞增殖能力明显降低,36h后与对照组相比[(1.71±0.17)vs.(1.39±0.11),P<0.05],72h后与对照组相比[(2.15±0.21)vs.(1.54±0.14),P<0.05],差异均有统计学意义;Tanswell实验显示转染pGenesil-STK31-shRNA后,MG63细胞迁移能力明显受到抑制[(13±4)个vs.(55±8)个,P<0.05]。结论 STK31在骨肉瘤组织中的表达升高,具有增加骨肉瘤细胞生物学活性的作用。Objective To observe the expression of serine/threonine kinase 31 （STK31） in osteosarcoma and its effect on the malignant biological behavior of osteosarcoma. Methods Fifteen cases of osteosarcoma specimens and adjacent normal tissue were collected. The expression of STK31 in tumor tissues and normal tissue were detected by immunohistochemistry,real-time quantita- tive PCR and Western blot. The STK31 knockout plasmids PGenesil-STK31-shRNA or control plasmid pGenesil-1 were transfected into osteosarcoma cell line MG63 cells. The effect of STK31 on the proliferation of MG63 cells was detected by CCK8 cell activity assay. Tanswell experiment was used to observed the effect of STK31 on the migration ability of osteosarcoma cells. Results Immunohistochemical showed that STK31 expressed in the tumor tissue,and it was significantly higher than the adjacent normal tissues;Real time quantitative PCR[（3. 65±0.83）vs. （1. 05±0.14） ,P〈0.05] and Western blot also revealed that STK31 expression in tumor tissue were significantly higher than adjacent normal tissues （P〈 0. 05）; CCK8 experiments showed that knockdown STK31 inhibited proliferation of MG63 cell when compared with the control group after 36 hi（1. 71±0.17）vs. （1. 39±0. 11）,P〈 0.05] ,72 hi（2. 15±0.21）vs. （1. 54±0.14）,P〈0.05] ;Tansewell experiments showed that transfection of pGenesil-STK31-shRNA could suppress MG63 cell＇s migration[（13±4）vs. （55±8） ,P,0.05]. Conclusion STK31 is overexpression in osteosarcoma with increased biological activity of osteosarcoma cells.

关 键 词：丝氨酸/苏氨酸激酶31 骨肉瘤 MG63 基因表达 

分 类 号：R738[医药卫生—肿瘤]