Department of Head and Neck Surgery, the Second Affiliated Hospital of Nanchang University, Nanchang, Jiangxi, 330006, China.
Department of General Surgery, People's Hospital of Jingan County, Yichun, Jiangxi, 330699, China.
Biomed Pharmacother. 2019 Dec;120:109508. doi: 10.1016/j.biopha.2019.109508. Epub 2019 Oct 22.
This study aimed to investigate the effect of interfering thyroid hormone receptor interacting protein 13 (TRIP13) expression on the proliferation, apoptosis and metastasis of thyroid cancer (TC) cells and the involved mechanisms. RT-PCR, immunohistochemical analysis and western blot found that compared with normal tissues, the expressions of TRIP13 and N-cadherin in TC tissues were significantly increased, while the expressions of tetratricopeptide repeat protein 5 (TTC5), p-p53 and E-cadherin were significantly decreased (P < 0.05). RT-PCR and western blot revealed that compared with C643 cells, TRIP13 expression in TPC1 cells and BHT101 cells increased significantly (P < 0.05). Therefore, TPC1 cells and BHT101 cells were selected for subsequent experiments. Interference efficiency of TRIP13 interference sequences (sh-TRIP13) was verified by RT-PCR and western blot. After sh-TRIP13 transfection, cell viability and migration rates of cells at 24 h and 48 h decreased significantly (P < 0.05); the number of S phase cells increased remarkably, while the number of G1 and G2 phase cells decreased significantly (P < 0.05); cell apoptosis was enhanced sharply (P < 0.05); cell numbers in the lower chamber of Transwell assay were reduced significantly (P < 0.05). RT-PCR and western blot found that compared with the Control group, expressions of TTC5 and E-cadherin in sh-TRIP13 group were elevated sharply, while N-cadherin expression decreased significantly (P < 0.05). Compared with the Control group, sh-TRIP13 transfection elevated the ratio of p-p53 expression to p53 expression (p-p53/p53) remarkably (P < 0.05). In conclusion, TRIP13 interference inhibited the proliferation and metastasis of thyroid cancer cells through regulating TTC5/p53 pathway and epithelial-mesenchymal transition related genes expression.
本研究旨在探讨干扰甲状腺激素受体相互作用蛋白 13（TRIP13）表达对甲状腺癌（TC）细胞增殖、凋亡和转移的影响及其机制。RT-PCR、免疫组织化学分析和 Western blot 发现，与正常组织相比，TC 组织中 TRIP13 和 N-钙黏蛋白的表达明显增加，而四肽重复蛋白 5（TTC5）、p-p53 和 E-钙黏蛋白的表达明显降低（P<0.05）。RT-PCR 和 Western blot 显示，与 C643 细胞相比，TPC1 细胞和 BHT101 细胞中 TRIP13 的表达明显增加（P<0.05）。因此，选择 TPC1 细胞和 BHT101 细胞进行后续实验。通过 RT-PCR 和 Western blot 验证了 TRIP13 干扰序列（sh-TRIP13）的干扰效率。sh-TRIP13 转染后，细胞在 24 h 和 48 h 的存活率和迁移率明显降低（P<0.05）；S 期细胞数量显著增加，而 G1 和 G2 期细胞数量明显减少（P<0.05）；细胞凋亡明显增强（P<0.05）；Transwell 小室下室的细胞数量明显减少（P<0.05）。RT-PCR 和 Western blot 发现，与对照组相比，sh-TRIP13 组的 TTC5 和 E-钙黏蛋白表达明显升高，而 N-钙黏蛋白表达明显降低（P<0.05）。与对照组相比，sh-TRIP13 转染明显提高了 p-p53 表达与 p53 表达的比值（p-p53/p53）（P<0.05）。综上所述，TRIP13 干扰通过调节 TTC5/p53 通路和上皮-间充质转化相关基因表达抑制甲状腺癌细胞的增殖和转移。
