多配体蛋白聚糖-1调控骨肉瘤细胞增殖和侵袭能力及机制研究  

Effects of syndecan-1 on proliferation and invasion against osteosarcoma cell and its mechanisms

作　　者：张翼[1] 张彬[1] 张岩[1] 卢新昌[1] 刘金磊 李甲振[1] Zhang Yi;Zhang Bin;Zhang Yan;Lu Xinchang;Liu Jinlei;Li Jiazhen(Department of Orthopedics,the First Affiliated Hospital of Zhengzhou University,Zhengzhou 450052,China)

机构地区：[1]郑州大学第一附属医院骨科,郑州450052

出　　处：《中华实验外科杂志》2024年第12期2791-2793,共3页Chinese Journal of Experimental Surgery

基　　金：河南省高等学校重点科研项目计划(23A320047)。

摘　　要：目的观察多配体蛋白聚糖-1(SDC1)调控骨肉瘤细胞增殖及侵袭能力并探讨其机制。方法通过SDC1小干扰RNA转染骨肉瘤MG-63细胞, 构建SDC1下调表达模型为实验组, 以转染阴性对照链作为对照组, 实时定量聚合酶链反应法验证SDC1 mRNA表达水平;比色法检测细胞增殖率和含半胱氨酸的天冬氨酸蛋白水解酶-3(Caspase-3)活性变化;流式细胞技术检测细胞凋亡水平变化;细胞体外侵袭试验检测细胞侵袭能力变化;蛋白质印迹法(Western blot)实验检测SDC1和基质金属蛋白酶-9(MMP-9)蛋白表达水平。组间比较采用独立样本t检验。结果 SDC1 siRNA转染后, MG-63细胞中SDC1 mRNA相对表达水平低于对照组(0.52±0.15比1.06±0.07, t=5.650, P<0.01);转染后24 h及48 h后siRNA组细胞增殖率均低于对照组(0.72±0.13比1.14±0.09、0.59±0.10比1.08±0.18, t=4.382、4.122, P<0.01);流式细胞检测结果显示siRNA组细胞凋亡率高于对照组[(9.53±1.97)%比(4.58±1.09)%, t=3.808, P<0.05], 而siRNA组Caspase-3相对活性高于对照组(1.96±0.27比1.05±0.19, t=4.774, P<0.01);体外侵袭实验证实siRNA组每视野下平均细胞数低于对照组[(76.22±12.02)个比(112.40±18.30)个, t=3.695, P<0.01];Western blot实验证实siRNA组细胞SDC1蛋白相对表达量低于对照组(0.56±0.24比1.07±0.10, t=3.411, P<0.05);SDC1组细胞MMP-9蛋白相对表达量低于对照组, 差异有统计学意义(0.62±0.11比0.97±0.14, t=3.405, P<0.05)。结论靶向下调SDC1表达可以抑制骨肉瘤细胞增殖和侵袭能力, 这与Caspase-3激活和下调MMP-9表达相关。ObjectiveTo investigate the effects of syndecan-1 on the proliferation and invasion against osteosarcoma cell and its mechanisms.MethodsThe expression of syndecan-1(SDC1)in osteosarcoma MG-63 cells was detected by Real-time quantitative polymerase chain reaction after SDC1 small interfering RNA(siRNA)transfection as the experimental group while the negative control transfection was set as the control group.After the validation of SDC1 mRNA down-expression,the colorimetric assays were used to detect the proliferation and cysteinyl aspartate specific proteinase-3(Caspase-3)activity;The apoptosis rate was detected by the flow cytometry assays.The Transwell assay was used to determine the invasion activity.Then the expression levels of SDC1 and matrix metalloprotein(MMP-9)were detected by Western blotting respectively.Independent sample t-test was used for comparison between two groups.ResultsAfter SDC1 siRNA transfection to the OS MG-63 cells,relative expression of SDC1 mRNA in siRNA group was significantly less than the negative-control(NC)group(0.52±0.15 vs.1.06±0.07,t=5.650,P<0.01).The proliferation at 24 h and 48 h in SDC1 siRNA group were lower than the NC group(0.72±0.13 vs.1.14±0.09,0.59±0.10 vs.1.08±0.18,t=4.382,4.122,P<0.01).Flow cytometry assay showed the apoptosis rate in siRNA group is higher than the NC group[(9.53±1.97)%vs.(4.58±1.09)%,t=3.808,P<0.05],while the relative activity of Caspase-3 was higher than NC group(1.96±0.27 vs.1.05±0.19,t=4.774,P<0.01).Transwell assay showed the invaded cells in siRNA group were significantly less than the NC group[(76.22±12.02)cells vs.(112.40±18.30)cells,t=3.695,P<0.01].After the transfection,the results from the Western-blot showed that the relative expression of SDC1 in siRNA group was lower than the NC group(0.56±0.24 vs.1.07±0.10,t=3.411,P<0.05),while the relative levels of MMP-9 in siRNA group was less than NC group(0.62±0.11 vs.0.97±0.14,t=3.405,P<0.05).ConclusionDown-regulation of SDC1 inhibited the proliferation,invasion of osteosarcoma MG-6

关 键 词：多配体蛋白聚糖-1 骨肉瘤 增殖 凋亡 侵袭 

分 类 号：R738.1[医药卫生—肿瘤]