Neoplasma. 2018;65(2):169-177. doi: 10.4149/neo_2018_170119N51.
Trefoil factor 3 (TFF3), a regulatory protein composed of 59 amino acids, has been suggested to be involved in pathogen- esis, proliferation, invasion, migration and apoptosis in multiple malignant tumors. However, the roles of TFF3 concerning the viability, migration and invasion in papillary thyroid carcinoma cells have not yet been studied. This study aimed to investigate the effect of TFF3 knockdown on a thyroid papillary carcinoma TPC-1 cell line both in vitro and in vivo. In the present study, lentivirus-mediated short hairpin RNA (shRNA) targeting TFF3 plasmids were first constructed and stable TPC-1 cells were obtained while their TFF3 gene was silenced with either shTFF3-TPC-1, or a scrambled shRNA control. TFF3 expression was detected using quantitative real-time PCR and western blot analyses. The TPC-1 cell viability was measured by CCK-8 assay and colony formation. The cell migration and invasion were assessed by wound scratch assay and transwell filters. AKT phosphorylation, MMP-9, and BCL-2 expression levels were detected by western blot analyses. Our results showed that TFF3 knockdown significantly inhibits TPC-1 cell viability, migration and invasion. AKT phosphoryla- tion, MMP-9, and BCL-2 levels were all remarkably depressed in TFF3 knockdown TPC-1 cells. Using a thyroid papillary carcinoma xenograft mouse model, we further investigated the effects of TFF3 knockdown in vivo. Significantly delayed xenograft emerging, slower growth rate and lower final tumor weights and volumes were observed in the shTFF3 group as compared to the control group. As expected, the expression levels of MMP-9 and BCL-2 in the xenograft are consistent with those of shTFF3-TPC-1 and shTFF3-TPC-1 cells in vitro. Our results suggest that TFF3 plays a vital role in the viability and oncogenesis of TPC-1 cells and may be a potential target for effective treatment of thyroid papillary carcinoma.
三叶因子 3（TFF3）是一种由 59 个氨基酸组成的调节蛋白，已被认为参与多种恶性肿瘤的发病机制、增殖、侵袭、迁移和凋亡。然而，TFF3 与甲状腺乳头状癌细胞的活力、迁移和侵袭的关系尚未得到研究。本研究旨在探讨 TFF3 敲低对甲状腺乳头状癌细胞系 TPC-1 的体内外作用。在本研究中，首先构建了靶向 TFF3 的慢病毒介导短发夹 RNA（shRNA）质粒，并获得了 TFF3 基因沉默的稳定 TPC-1 细胞，即 shTFF3-TPC-1 或乱序 shRNA 对照。采用实时定量 PCR 和 Western blot 分析检测 TFF3 表达。通过 CCK-8 测定和集落形成实验检测 TPC-1 细胞活力。通过划痕实验和 Transwell 滤器评估细胞迁移和侵袭。通过 Western blot 分析检测 AKT 磷酸化、MMP-9 和 BCL-2 的表达水平。结果显示，TFF3 敲低显著抑制 TPC-1 细胞活力、迁移和侵袭。TFF3 敲低的 TPC-1 细胞中 AKT 磷酸化、MMP-9 和 BCL-2 水平显著降低。在甲状腺乳头状癌异种移植小鼠模型中，我们进一步研究了 TFF3 敲低的体内作用。与对照组相比，shTFF3 组的异种移植出现明显延迟，生长速度较慢，最终肿瘤重量和体积较低。正如预期的那样，异种移植中的 MMP-9 和 BCL-2 表达水平与 shTFF3-TPC-1 和 shTFF3-TPC-1 细胞在体外的表达水平一致。研究结果表明，TFF3 在 TPC-1 细胞的活力和肿瘤发生中起着重要作用，可能是治疗甲状腺乳头状癌的有效靶点。
