GPX3过表达对甲状腺乳头状癌细胞增殖、迁移的影响

Effect of overexpression of GPX3 on proliferation and migration of thyroid papillary carcinoma cells

目的探讨谷胱甘肽过氧化物酶3(glutathione peroxidase 3,GPX3)过表达对甲状腺乳头状癌细胞(BCPAP)增殖、迁移的影响。方法培养人甲状腺乳头状癌BCPAP细胞,利用MOI值为10的过表达GPX3慢病毒和空载慢病毒转染BCPAP细胞株,72 h后用共聚焦显微镜观察慢病毒转染细胞的效率,并分为二组:空载转染组(NC-BCPAP组)和GPX3转染组(GPX3-BCPAP组);qRT-PCR检测GPX3表达以及Caspase-3表达情况;CCK-8法及BeyoClick EdU-555检测细胞增殖情况;划痕实验检测细胞的迁移能力。用SPSS 22.0统计软件分析处理,实验重复3次。结果转染72 h后利用共聚焦显微镜观察BCPAP细胞中绿色荧光证明转染成功。采用qRT-PCR检测表明GPX3-BCPAP组细胞的GPX3表达明显高于NC-BCPAP组(P<0.001),与NC-BCPAP组细胞相比,GPX3-BCPAP组细胞中Caspase-3表达水平较高,表明过表达GPX3可能促进细胞的凋亡能(P<0.005)。采用CCK-8及BeyoClick EdU-555检测表明与NC-BCPAP组细胞相比,GPX3-BCPAP组细胞的增殖能力受到明显抑制(P<0.05)。采用划痕实验表明与NC-BCPAP组细胞相比,GPX3-BCPAP组细胞的迁移能力受到明显抑制(P<0.05)。结论GPX3在抑制BCPAP细胞的增殖、迁移起到关键作用。

Objective To investigate the effect of glutathione peroxidase 3(glutathione peroxidase 3,GPX3)overexpression on the proliferation and migration of thyroid papillary carcinoma cell line(BCPAP).Methods To investigate the effect of glutathione peroxidase 3(glutathione peroxidase 3,GPX3)overexpression on the proliferation and migration of thyroid papillary carcinoma cell line(BCPAP).Methods Human papillary thyroid carcinoma BCPAP cells were cultured and transected into BCPAP cell line with over-expressed GPX3 lentivirus and no-load lentivirus with MOI 10.72 hours later,the efficiency of lentivirus transfection was observed by confocal microscope and divided into two groups:no-load transfection group(NC-BCPAP group)and GPX3 transfection group(GPX3-BCPAP group).The expression of GPX3 and Caspase-3 was detected by qRT-PCR.The proliferation of cells was detected by CCK-8 and BeyoClick EdU-555,and the migration ability of cells was detected by scratch test.The SPSS 22.0 analyzed and processed experiment statistical software,and the experiment was repeated 3 times.Results 72 hours after transfection,the green fluorescence in BCPAP cells was observed by confocal microscope to prove that the transfection was successful.qRT-PCR detection showed that the expression of GPX3 in GPX3-BCPAP group was significantly higher than that in NC-BCPAP group(P<0.001).Compared with NC-BCPAP group,the expression level of Caspase-3 in GPX3-BCPAP group was higher,indicating that overexpression of GPX3 may promote cell apoptosis(P<0.005).The Results of CCK-8 and BeyoClick EdU-555 showed that the proliferation of cells in GPX3-BCPAP group was significantly inhibited compared with that in NC-BCPAP group(P<0.05).The scratch test showed that the migration ability of cells in NC-BCPAP group was significantly inhibited compared with that in GPX3-BCPAP group(P<0.05).Conclusion GPX3 plays a key role in inhibiting the proliferation and migration of BCPAP cells.