SEMA6D对人骨肉瘤MG63细胞生物学行为的影响  

Effect of SEMA6D on biological behavior of human osteosarcoma MG63 cells

作　　者：都展鸿 杨维经 王雨婵 付雅璐 李庆慧 孟玉菡 刘兴龙 DU Zhan-hong;YANG Wei-jing;WANG Yuchan;FU Ya-lu;LI Qing-hui;MENG Yu-han;LIU Xing-long(Affiliated Hospital,Shandong Second Medical University,Weifang,261000 Shandong,China)

机构地区：[1]山东第二医科大学附属医院,山东潍坊261000

出　　处：《中国矫形外科杂志》2025年第6期535-540,共6页Orthopedic Journal of China

基　　金：潍坊市科技发展计划项目(编号:2022YX032);山东省学校卫生协会重点课题(编号:JKZX2023136)。

摘　　要：[目的]探讨信号素6D(semaphorin 6D,SEMA6D)对骨肉瘤细胞增殖、侵袭的影响及其机制。[方法]将MG63细胞分为空白对照组(Ctrl组)、阴性对照组(si-NC组)和转染靶向SEMA6D的si-RNA组(si-SEMA6D组),给予相应的体外转染。采用CCK-8、细胞划痕、Transwell侵袭实验检测MG63细胞增殖、迁移及侵袭能力的变化。Western blot检测下游相关信号通路蛋白表达情况。[结果]培养24 h,三组CCK-8值差异无统计学意义(P>0.05),在48 h和72 h,Ctrl、si-NC组CCK-8值显著高于si-SEMA6D组[(0.7±0.1)vs(0.7±0.1)vs(0.4±0.1),P<0.001;(1.7±0.1)vs(1.6±0.1)vs(1.0±0.1),P<0.001]。Transwell侵袭实验显示,Ctrl、si-NC组细胞侵袭数显著高于si-SEMA6D组[(435.0±28.2)vs(400.7±41.4)vs(291.3±31.1),P=0.022]。划痕24 h、48 h,Ctrl、si-NC组划痕愈合率显著高于si-SEMA6D组[(48.8±3.3)%vs(40.6±3.4)%vs(16.6±2.4)%,P<0.001;(74.7±1.1)%vs(67.6±3.0)%vs(49.5±2.3)%,P<0.001]。Western blot检测表明,相较于si-SEMA6D组,Ctrl组、si-NC组p-PI3K、p-AKT、pp38、MMP2的蛋白表达水平均显著增加(P<0.05),而三组中P13K、AKT、p38、Bcl-2及Bax的表达水平差异无统计学意义(P>0.05)。[结论]沉默SEMA6D后骨肉瘤MG63细胞可通过抑制PI3K/AKT及p38-MAPK信号通路,抑制细胞增殖、迁移及侵袭能力。[Objective]To investigate the effects of semaphorin 6D(SEMA6D)on the proliferation and invasion of osteosarcoma cells and its mechanism.[Methods]MG63 cells were divided into blank control group(the Ctrl group),negative control group(the si-NC group)and the group transfected with si-RNA targeting SEMA6D(the si-SEMA6D group),and corresponding transfection was performed in vitro.The changes of proliferation,migration and invasion ability of MG63 cells were detected by CCK-8,cell scratch and Transwell invasion assay.In addition,western blot analysis was performed to detect the protein expression of downstream related signaling pathways.[Results]After 24 hours culture,there was no significant difference in CCK-8 among the three groups(P>0.05).At 48 hours and 72 hours,the Ctrl and si-NC groups were significantly higher than the si-SEMA6D groups in CCK-8 assay[(0.7±0.1)vs(0.7±0.1)vs(0.4±0.1),P<0.001;(1.7±0.1)vs(1.6±0.1)vs(1.0±0.1),P<0.001].The Ctrl and si-NC groups were significantly higher than the si-SEMA6D group in Transwell invasion assay[(435.0±28.2)vs(400.7±41.4)vs(291.3±31.1),P=0.022].At 24 hours and 48 hours,the Ctrl and si-NC groups had significantly higher scratch healing rate than the si-SEMA6D group[(48.8±3.3)%vs(40.6±3.4)%vs(16.6±2.4)%,P<0.001;(74.7±1.1)%vs(67.6±3.0)%vs(49.5±2.3)%,P<0.001].As consequence of western blot analysis,the Ctrl group and si-NC group were significantly increased compared with the si-SEMA6D group in protein expression levels of p-PI3K,p-AKT,p-P38 and MMP2(P<0.05),although there were no significant differences in the expression levels of P13K,AKT,p38,Bcl-2 and Bax among the three groups(P>0.05).[Conclusion]SEMA6D silencing of osteosarcoma MG63 cells can inhibit cell proliferation,migration and invasion by inhibiting PI3K/AKT and p38-MAPK signaling pathways.

关 键 词：骨肉瘤 SEMA6D 侵袭 PI3K/AKT 

分 类 号：R738.1[医药卫生—肿瘤]