摘要目的 观察盐诱导激酶1(SIK1)对肝癌细胞增殖、侵袭和迁移的调控作用.方法 将抑制剂转染肝癌MHCC97-L和HEP3B细胞株构建SIK1敲低组,通过实时聚合酶链反应(qPCR)检测SIK1的表达;噻唑蓝(MTT)法检测SIK1在1、2、3、4、5d时对细胞增殖的影响;细胞周期实验检测SIK1对细胞G1、S、G2期的影响;细胞凋亡实验检测细胞侵袭能力;划痕实验检测6、24、48 h时细胞的迁移能力.结果 SIK1在肝癌MHCC97-L和HEP3B细胞中相对表达低于正常肝脏细胞(0.51±0.01、0.33±0.02,tMHCC97-L=19.630,P=0.000;tHEP3B=24.940,P=0.000);转染SIK1抑制物后,肝癌MHCC97-L和HEP3B细胞内SIK1相对表达量明显降低(0.39±0.02、0.51±0.03,tMHCC97-L=11.479,P=0.000;tHEP3B=9.543,P=0.001);敲低SIK1后,MTT实验结果显示,明显促进MHCC97-L和HEP3B细胞的增殖(1.20±0.06比0.90±0.07,tMHCC97-L=5.630,P=0.005;1.09±0.11比0.63±0.05,tHEP3B=6.460,P=0.003);细胞周期实验结果显示:S期细胞比例高于对照组[(24.77±0.99)％比(17.22±1.01)％,tMaCC97-L=9.280,P=0.001;(21.63±0.94)％比(15.32±0.88)％,tHEP3B=8.460,P=0.001];G2细胞比例高于对照组[(16.75±0.50)％比(14.94±0.64)％,tMHCC97-L=3.860,P=0.018;(15.99±0.51)％比(13.97±0.44)％,tHEP3B=5.170,P=0.007],促进细胞从G1期向S期转变;细胞凋亡实验显示:MHCC97-L和HEP3B细胞凋亡率下降[(1.91±0.10)％比(3.27±0.31)％,tMHCC97-L =7.180,P=0.002;(2.01±0.14)％比(3.81±0.13)％,tHEP3B=16.600,P =0.000];划痕实验结果显示:明显促进MHCC97-L和HEP3B细胞迁移[(202.00±15.61)比(285.33±10.07) μm、(229.33±13.61)比(321.33±16.04) μm,tMHCC97-L =7.770,P =0.001;tHEP3B=7.570,P=0.002],差异有统计学意义.结论 降低SIK1的表达促进了MHCC97-L和HEP3B细胞株增殖、侵袭和迁移的作用.

abstractsObjective To investigate the effect of salt-induced kinase 1 (SIK1) on the proliferation,invasion and migration of hepatoma cells.Methods Inhibitors were transfected into MHCC97-L and HEP3B cells to construct SIK1 knockdown group.The expression of SIK1 was detected by quantitative polymerase chain reaction (qPCR).The effect of SIK1 on cell proliferation at 1,2,3,4,5 days was measured by MTT assay.Cell cycle assay was used to detect the effect of SIK1 on G1,S and G2 phases.Apoptosis assay was used to detect cell invasion.The migration of osteosarcoma cells at 6,24,48 h was determined by wound healing test.Results The expression of SIK1 in MHCC97-L and HEP3B cells was lower than that in normal liver cells (0.51 ± 0.01,0.33 ± 0.02,tMHCC97-L =19.630,P =0.000;tHEP3B =24.940,P =0.000).After knocking down SIK1,the expression of SIK1 in MHCC97-L and HEP3B cells was significantly decreased(0.39 ± 0.02,0.51 ± 0.03,tMHCC97-L =11.479,P =0.000;tHEP3B =9.543,P =0.001).MTT assay showed that the proliferation of MHCC97-L and HEP3B cells was significantly improved (1.20 ±0.06 vs.0.90 ±0.07,tMHCC97-L =5.630,P=0.005;1.09 ±0.11 vs.0.63 ±0.05,tHEP3B =6.460,P =0.003);Cell cycle experiments showed that the ratio of S phase cells was higher than that of the control group [(24.77 ± 0.99) ％ vs.(17.22 ± 1.01) ％,tMHCC97-L =9.280,P =0.001;(21.63 ±0.94)％ vs.(15.32 ±0.88)％,tHEP3B =8.460,P =0.001],and the ratio ofG2 cells was higher than that of the control group [(16.75 ± 0.50) ％ vs.(14.94 ± 0.64) ％,tMHCC97-L =3.860,P =0.018;(15.99 ± 0.51) ％ vs.(13.97 ± 0.44) ％,tHEP3B =5.170,P =0.007].The apoptosis rate of MHCC97-L and HEP3B cells was decreased [(1.91 ± 0.10) ％ vs.(3.27 ± 0.31) ％,tMHCC97-L =7.180,P=0.002;(2.01 ±0.14)％ vs.(3.81 ±0.13)％,tHEP3B =16.600,P=0.000].The results of wound healing test showed that the migration of MHCC97-L and HEP3B cells was promoted [(202.00 ± 15.61)vs.(285.33 ± 10.07) μm;(229.33 ± 13.61) vs.(321.33 ± 16.04) μm,tMHCC97-L=7.770,P=0.001;tHEP3B =7.570,P =0.002].Conclusion Decreasing the expression of SIK1 promotes the proliferation,invasion and migration of MHCC97-L and HEP3B cells.