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目的:构建CGTHW-1/pGenesil-1-CD151shRNA稳转细胞系和空载体细胞系CGTHW-1/pGenesil-1,探讨CD151对人甲状腺癌CGTHW-1细胞迁移及侵袭的影响及其作用机制。方法:应用Lipofectamine2000将真核干扰载体pGenesil-1-CD 151 shRNA和空载体pGenesil-1导入甲状腺滤泡癌细胞CGTHW-1,经卡那霉素抗性筛选得到稳定的克隆并扩大培养成细胞系,Western blot检测CD151在CGTHW-1细胞中的表达;划痕实验和Transwell实验分别观察CD151对细胞迁移和侵袭能力的影响。结果:成功建立了CGTHW-1/pGenesil-1-CD151shRNA稳转细胞系和空载体细胞系CGTHW-1/pGenesil-1,Western blot结果显示CD151在CGTHW-1细胞中表达,干扰CD151基因后,CGTHW-1细胞的迁移和侵袭能力明显降低。结论:将CD151基因干扰后可明显抑制甲状腺癌细胞的迁移和侵袭能力。CD151可能是影响甲状腺癌侵袭转移的重要因子之一。
OBJECTIVE: To construct CGTHW-1 / pGenesil-1 cell line CGTHW-1 / pGenesil-1-CD151shRNA stable transfected and CGTHW-1 / pGenesil-1 to investigate the effect of CD151 on migration and invasion of human thyroid carcinoma CGTHW-1 cells and its mechanism. Methods: The eukaryotic expression vector pGenesil-1-CD151 and empty vector pGenesil-1 were transfected into CGTHW-1 cells by lipofectamine 2000. The stable clones were screened by kanamycin resistance and expanded into cell lines The expression of CD151 in CGTHW-1 cells was detected by Western blot. The scratch test and Transwell assay were used to observe the effect of CD151 on the migration and invasion of cells. Results: CGTHW-1 / pGenesil-1-CD151shRNA stable cell line and empty vector cell line CGTHW-1 / pGenesil-1 were successfully established. Western blot results showed that CD151 was expressed in CGTHW-1 cells. -1 cells significantly reduced the ability of migration and invasion. Conclusion: CD151 gene interference can significantly inhibit the migration and invasion of thyroid cancer cells. CD151 may be one of the important factors affecting the invasion and metastasis of thyroid cancer.