Stathmin基因在^(131)I抑制甲状腺癌细胞生长中的作用

The role of stathmin in^(131)I inhibiting the growth of thyroid cancer cells

目的探讨微管不稳定蛋白(stathmin)基因在^(131)I抑制甲状腺癌细胞生长中的作用及相关机制。方法将^(131)I抵抗甲状腺癌细胞株(res⁃TPC⁃1)随机分为对照组、^(131)I组、si⁃NC组、si⁃stathmin组、^(131)I+si⁃NC组和^(131)I+si⁃stathmin组。采用MTT实验检测res⁃TPC⁃1细胞存活率。克隆形成实验检测克隆形成数。流式细胞术检测凋亡率。TUNEL染色检测凋亡指数。Transwell检测迁移数。Western blot检测stathmin、细胞周期蛋白D1(CyclinD1)、细胞周期蛋白依赖性激酶抑制剂1A(p21)、B淋巴细胞瘤-2相关X蛋白(Bax)、B淋巴细胞瘤-2(Bcl⁃2)、基质金属蛋白酶-2(MMP⁃2)、基质金属蛋白酶-9(MMP⁃9)、β-连环蛋白(β-catenin)和c⁃Myc蛋白表达水平。结果Stathmin在res⁃TPC⁃1细胞中的表达水平明显高于TPC⁃1细胞,差异有统计学意义(t=-18.552,P<0.05)。与对照组比较,^(131)I组res⁃TPC⁃1细胞存活率、克隆形成数、迁移数、CyclinD1、Bcl⁃2、MMP⁃2和MMP⁃9水平明显降低,差异有统计学意义(t值分别为3.786、6.293、5.226、5.979、6.882、6.578和4.500,P<0.05),凋亡率、凋亡指数、p21、Bax、β-catenin和c⁃Myc水平明显增加,差异有统计学意义(t值分别为-8.131、-11.496、-6.240、-6.600、-13.950和-10.262,P<0.05)。与si⁃NC组比较,si⁃stathmin组res⁃TPC⁃1细胞存活率、克隆形成数、迁移数、CyclinD1、Bcl⁃2、MMP⁃2、MMP⁃9、β-catenin和c⁃Myc水平明显降低,差异有统计学意义(t值分别为3.793、9.199、9.193、10.733、9.430、9.907、7.496、9.261和12.862,P<0.05),凋亡率、凋亡指数、p21和Bax水平明显增加,差异有统计学意义(t值分别为-12.675、-14.712、-6.626和-6.091,P<0.05)。与^(131)I+si⁃NC组比较,^(131)I+si⁃stathmin组res⁃TPC⁃1细胞存活率、克隆形成数、迁移数、CyclinD1、Bcl⁃2、MMP⁃2、MMP⁃9、β-catenin、c⁃Myc水平明显降低,差异有统计学意义(t值分别为7.134、11.324、8.589、8.558、10.817、10.401、11.833、10.490和9.098,P<0.05),凋亡率、凋亡指数、p21、Bax水平明显增加,差异有统计学意义(t值分别为-13.190、-13.284、-10.817和-10.419,P<0.05)。结论抑制stathmin表达可能通过抑制Wnt/β-catenin信号通路增强^(131)I对res⁃TPC⁃1细胞的杀伤效果。

Objective To explore the role and mechanism of stathmin in^(131)I inhibiting the growth of thyroid cancer cells.Methods^(131)I resistant thyroid cancer cell line Res⁃TPC⁃1 were randomly divided into control group,^(131)I group,si⁃NC group,si⁃stathmin group and^(131)I+si⁃NC group,^(131)I+si⁃stathmin group.MTT was used to detect the survival rate of res⁃TPC⁃1 cells.Clonal formation assay was used to detect the clonal formation number.Flow cytometry was used to detect the apoptosis rate.TUNEL staining was used to detect the apoptotic index.Transwell was used to detect the cell migration level.Western blot was used to detect the protein expression level of stathmin,CyclinD1,cyclin⁃dependent kinase inhibitors 1A(p21),B⁃lymphocytoma⁃2-associated X protein(Bax),B⁃lymphocytoma⁃2(Bcl⁃2),matrix metalloproteinase⁃2(MMP⁃2),matrix metalloproteinase⁃9(MMP⁃9),beta⁃catenin(β-catenin)and c⁃Myc.Results The expression level of stathmin in res⁃TPC⁃1 cells was significantly higher than that in TPC⁃1 cells,the difference was statistically significant(t=-18.552,P<0.05).Compared with the control group,the survival rate,clonal formation number,migration number,levels of CyclinD1,Bcl⁃2,MMP⁃2 and MMP⁃9 of res⁃TPC⁃1 cells in^(131)I group were significantly decreased,the differences were statistically significant(t values were 3.786,6.293,5.226,5.979,6.882,6.578,4.500,respectively,P<0.05),while the apoptosis rate,apoptosis index,and the levels of p21,Bax,β-catenin and c⁃Myc were significantly increased,and the differences were statistically significant(t values were-8.131,-11.496,-6.240,-6.600,-13.950,-10.262,respectively,P<0.05).Compared with the si⁃NC group,the survival rate,clonal formation number,migration number,levels of CyclinD1,Bcl⁃2,MMP⁃2,MMP⁃9,β-catenin and c⁃Myc of res⁃TPC⁃1 cells in si⁃stathmin group were significantly decreased,the differences were statistically significant(t values were 3.793,9.199,9.193,10.733,9.430,9.907,7.496,9.261,12.862,respectively,P<0.05),while the apoptosis rate,apoptosis index,the levels of p21 and Bax were significantly increased,the differences were statistically significant(t values were-12.675,-14.712,-6.626,-6.091,respectively,P<0.05).Compared with the 131 I+si⁃NC group,the survival rate,clonal formation number,migration number,levels of CyclinD1,Bcl⁃2,MMP⁃2,MMP⁃9,β-catenin and c⁃Myc of res⁃TPC⁃1 cells in131 I+si⁃stathmin group were significantly decreased,the differences were statistically significant(t values were 7.134,11.324,8.589,8.558,10.817,10.401,11.833,10.490,9.098,respectively,P<0.05),while the apoptosis rate,apoptosis index,the levels of p21 and Bax were significantly increased,the differences were statistically significant(t values were-13.190,-13.284,-10.817,-10.419,respectively,P<0.05).Conclusion Inhibition of stathmin expression may enhance the killing effect of131 I on res⁃TPC⁃1 cells by inhibiting Wnt/β-catenin signaling pathway.