文章摘要
骨桥蛋白对甲状腺癌细胞生物学效应的影响
Effect of osteopontin on biological effects of thyroid cancer cells
投稿时间：2018-08-29  
DOI：10.3969/j.issn.1000-0399.2019.04.002
中文关键词: 骨桥蛋白  甲状腺癌  凋亡  侵袭
英文关键词: Osteopontin  Thyroid carcinoma  Apoptosis  Invasion
基金项目:
作者单位
李豪杰 462300 河南省漯河市郾城区人民医院外二科 
徐庭凯 462300 河南省漯河市郾城区人民医院外二科 
王连迪 462300 河南省漯河市郾城区人民心内科 
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中文摘要:
      目的 探讨骨桥蛋白（OPN）对甲状腺癌生物学行为的影响。方法 SW579细胞置于6孔板中培养，采用随机数字表法分成对照组、阴性对照组、OPN siRNA干扰组3组，每组3个孔，每个孔2 mL，细胞密度为1×104/mL。对照组不作任何处理，阴性对照组转染OPN阴性序列，OPN siRNA干扰组转染OPN siRNA序列。采用实时荧光定量逆转录聚合酶链反应（qRT-PCR）测定并比较转染后3组细胞OPN、MMP-2、MMP-9、Caspase-3的mRNA水平，四甲基偶氮唑盐（MTT）测定转染后3组细胞的抑制率，流式细胞仪检测转染后3组细胞凋亡率，Transwell小室检测转染后3组细胞的侵袭力，蛋白免疫印迹法（Western blot）检测3组OPN、MMP-2、MMP-9、Casepase-3的蛋白水平。结果 OPN siRNA干扰组的OPN mRNA的表达水平显著低于对照组和阴性对照组，细胞凋亡率显著高于对照组以及阴性对照组，侵袭能力显著低于对照组和阴性对照组，OPN、MMP-2、MMP-9 mRNA和蛋白水平显著低于对照组和阴性对照组，Casepase-3mRNA和蛋白水平显著高于对照组和阴性对照组，差异均有统计学意义（P<0.05）。结论 OPN干扰后SW579细胞增殖和侵袭能力下降，凋亡率增加，其机制可能与凋亡蛋白caspase-3水平增加，MMP-2、MMP-9水平下降有关。
英文摘要:
      Objective To investigate the effect of osteopontin (OPN) on the biological behavior of thyroid cancer. Methods SW579 cells were cultured in 6-well plate and randomly divided into control group, negative control group and OPN small interfering RNA (siRNA) interference group, with 2×104 cells in each group, 3 replicates in each group. The control group did not receive any treatment, the negative control group was transfected with OPN negative sequence, and OPN siRNA interference group was transfected with OPN siRNA sequence. qRT-PCR was used to determine the mRNA levels of OPN, MMP-2, MMP-9 and Caspase-3 in each group after transfection. The inhibitory rate of each group after transfection was detected by MTT. The apoptosis of each group after transfection was detected by flow cytometry. The invasiveness of each group was detected by Transwell chamber after transfection. The protein levels of OPN, MMP-2, MMP-9 and Casepase-3 were detected by Western blot. Results The expression of OPN mRNA in OPN siRNA interference group was significantly lower than that in control group and negative control group (P<0.05), and the apoptotic rate was significantly higher than that in control group and negative control group (P<0.05). The invasion ability was significantly lower than that in control group and negative control group (P<0.05). The mRNA and protein levels of MMP-2 and MMP-9 were significantly lower than those of control group and negative control group, and the levels of Casepase-3 mRNA and protein were significantly higher than those of control group and negative control group (P<0.05). Conclusion After OPN interference, the proliferation and invasion ability of SW579 cells decreased, while the apoptotic rate increased significantly. The mechanism may be related to the increase of apoptotic protein caspase-3 level and the decrease of MMP-2 and MMP-9 levels.
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