微小RNA-1243直接靶向丝氨酸／苏氨酸蛋白激酶1和丝氨酸／苏氨酸蛋白激酶2调节甲状腺乳头状癌TPC-1细胞迁移  

MicroRNA-1243 regulates cell migration by targeting serine/threonine protein kinase 1 and serine/threonine protein kinase 2 in thyroid papillary carcinoma TPC-1 cell

作　　者：冀宏[1] 底旺[1] 李清怀[1] Ji Hong;Di Wang;Li Qinghuai(Department of Thyroid Surgery,the Second Hospital of Hebei Medical University,Shifiazhuang 050000,China)

机构地区：[1]河北医科大学第二医院甲状腺外科,石家庄050000

出　　处：《中华实验外科杂志》2018年第11期2048-2050,共3页Chinese Journal of Experimental Surgery

摘　　要：目的观察微小RNA（miRNA，miR）-1243调节丝氨酸／苏氨酸蛋白激酶1（Akt1）和丝氨酸／苏氨酸蛋白激酶2（Akt2）在人甲状腺乳头状癌细胞TPC-1中的表达及对细胞迁移的影响。方法利用TargetScan和miRanda对人甲状腺乳头状癌TPC-1细胞株的miR-1243靶基因进行预测，采用双荧光素酶报告基因验证miR-1243对靶基因的直接调控。Western blot检测miR-1243对Akt1和Akt2蛋白表达的调节。迁移小室（Transwell）检测1×10^5个对数生长期的TPC-1细胞的迁移能力。结果TargetScan和miRanda软件预测显示Akt1和Akt2基因是miR-1243的潜在靶基因。双荧光报告基因的相对荧光值显示，与空载体组和突变组比较，分别共转染miR-1243模拟物的Akt1或Akt2野生型组荧光素酶活性显著下降（t=3.595，P=0.009），而空载体组和突变组的荧光素酶活性差异无统计学意义（t=1.655，P=0.213）。与对照处理组比较，miR-1243模拟物处理组显著抑制而miR-1243抑制物处理组促进TPC-1细胞Akt1和Akt2蛋白水平的表达（t=4.810，P=0.018），而内参基因蛋白水平均无明显变化（P=0.235）。Transwell实验结果显示，miR-1243模拟物转染组迁移细胞数[（9.83±3.51）个]低于对照模拟物处理组[（18.67±4.24）个]，差异均有统计学意义（t=2.692，P=0.039）。与对照抑制物处理组比较[（21.33±5.35）个]，miR-1243抑制物转染组迁移细胞数[（37.17±7.33）个]显著增加（t=2.472，P=0.022）。结论miR-1243通过靶向抑制Akt1和Akt2的表达进而调节TPC-1细胞迁移。Objective To study microRNA （miRNA, miR）-1243 direct targeting serine/threonine protein kinase 1（Aktl） and serine/threonine protein kinase 2 （Akt2） regulation of thyroid papillary carcinoma TPC-1 cell migration. Methods miR-1243 target gene was predicted by TargetScan and mi-Randa. The double luciferase reporter gene was used to verify the direct regulation of miR-1243 on target gene. Western blotting was used to detect the protein level of Aktl and Akt2 in miR-1243 transfected cell. The migration of cells was detected by transwell. Results The TargetScan and miranda software predict that the Aktl and Akt2 genes are potential target genes of miR-1243. The relative fluorescence values of the double fluorescent reporter gene showed that the activity of luciferase in the Aktl or Akt2 wild-type group transfected with miR-1243 mimics was significantly lower than that of the empty vector group and the mutant group （t =3. 595, P =0. 018）. There was no significant difference in lueiferase ac- tivity between the vector group and the mutant group （t =1.655, P =0.213）. Compared with the control group, the miR-1243 mimic treatment group was significantly inhibited and the miR-1243 inhibitor treatment group promoted the expression of Aktl and Akt2 protein in TPC-1 cells （ t=4. 810, P=0.009） , but the protein level of the internal reference gene was not changed （P = 0.235）. The results of transwell showed that the number of migratory cells in the miR-1243 mimic transfected group [（9.83±3.51） cells] was lower than that in the control group [（18.67±4. 24） cells] , the difference was statisti-cally significant （t= 2.692, P=0.039）. Compared with the control inhibitor group [（21.33±5.35） cells], the number of migrating cells in the miR-1243 inhibitor transfeetion group [ （37.17±7.33 ） cells ] was significantly increased （t=2.472, P=0. 022）. Conclusion miR-1243 can inhibit the expression of Aktl and Akt2 and regulate the migration of TPC-1 cells.

关 键 词：TPC-1细胞 微小RNA-1243 细胞迁移 丝氨酸/苏氨酸蛋白激酶1丝氨 酸/苏氨酸蛋白激酶2 

分 类 号：R736.1[医药卫生—肿瘤]