基因沉默LMTK3对甲状腺乳头状癌细胞株BCPAP恶性生物学行为的影响

The effects of gene silencing LMTK3 on the malignant biological behaviors of human thyroid papillary cancer cell line BCPAP

目的探讨基因沉默LMTK3对人甲状腺乳头状癌细胞株BCPAP增殖、迁移、侵袭和凋亡等恶性生物学行为的影响。方法用携带sh LMTK3的重组慢病毒载体(LV-sh LMTK3-1、LV-sh LMTK3-2和LV-sh LMTK3-3)分别感染人甲状腺乳头状癌BCPAP细胞株,作为实验组(LV-sh LMTK3组);以对照慢病毒载体(LV-Control)感染人甲状腺乳头状癌BCPAP细胞株,作为阴性对照组(LV-Control组);空白对照组(Control组)细胞常规培养,不作任何处理。用Western Blot检测各组细胞中LMTK3蛋白的表达;CCK8实验检测细胞增殖能力的变化;Transwell实验检测细胞迁移和侵袭能力的变化;流式细胞术检测各组细胞的凋亡能力。结果 Western blot方法显示LV-sh LMTK3组中LMTK3蛋白的表达较阴性对照组显著降低(P<0.01);CCK8实验显示基因沉默LMTK3显著抑制BCPAP细胞的增殖(P<0.001);Transwell实验结果显示,与LV-Control组相比,LV-sh LMTK3组细胞的迁移和侵袭能力均明显降低(均为P<0.01),流式细胞术显示基因沉默LMTK3明显促进细胞的凋亡(P<0.01)。结论基因沉默LMTK3可显著抑制BCPAP细胞的增殖、迁移和侵袭能力,促进其凋亡。

Objective To investigate the effects of gene silencing LMTK3 on malignant biological behaviors of human thyroid papillary cancer cell line BCPAP,includingcell proliferation,migration,invasion and apoptosis.Methods The experimental group（ LV-sh LMTK3 group） is that thyroid papillary cancer BCPAP cells were transfected with recombinant lentivirus vectors（ LV-sh LMTK3-1,LV-sh LMTK3-2 and LV-sh LMTK3-3）. The negative control group（ LV-Control） was transfected with negative control lentiviral vector,and the blank control group（ Control） without any treatment. The protein expression of LMTK3 in cells was validated by western blot. Proliferation of the cells was detected by CCK8 assay. Transwell assays were used to determine the cells migration and invasion. Apoptosis of the cells was analyzed by flow cytometry. Results Western blot demonstrated that the LMTK3 protein expression was markedly decreased in the LV-sh LMTK3 group compared with the LV-control groups（ P〈0. 01）. CCK8 assay revealed that gene silencing of LMTK3 markedly reduced BCPAP cellular proliferation activity（ P〈0. 001）. Transwell assays showed that the migration and invasion of BCPAP cells were decreased significantly in the LV-sh LMTK3 group compared with the LV-Control group（ all P〈0. 01）. And flow cytometry displayed that gene silencing of LMTK3 promoted the apoptosis rate（ P〈0. 01）.Conclusion Gene silencing of LMTK3 significantly depresses human thyroid papillary cancer BCPAP cells proliferation,migration,and invasion and induces apoptosis.