Zhang Xing-Guang, Lu Xiao-Feng, Jiao Xiu-Ming, Chen Bin, Wu Jin-Xiao
Department of Endocrinology, The Military General Hospital of Beijing PLA, Beijing 100700, P.R. China.
Exp Ther Med. 2012 Dec;4(6):1005-1009. doi: 10.3892/etm.2012.729. Epub 2012 Sep 28.
The aim of this study was to observe the regulatory action of the polo-like kinase 1 (PLK1) gene in the invasion of anaplastic thyroid carcinoma cells and investigate its mechanisms. The expression of the PLK1 protein in 36 patients with anaplastic thyroid carcinoma was detected by immunohistochemical staining. siRNA against PLK1 was designed, synthesized and transfected into ARO cells. The effects of PLK1 siRNA on cell invasion were detected by a soft agar colony formation assay and a Transwell chamber assay. The corresponding protein was detected using western blot analysis. The expression of PLK1 in anaplastic thyroid carcinoma samples (67.5±10.6%) was significantly higher compared to that in cancer-adjacent samples (0.65%±0.12%; P<0.01). The expression of PLK1 correlated with clinical stage, lymph node metastasis and prognosis of anaplastic thyroid. The number of cell clones was reduced in a dose-dependent manner with increasing levels of siRNA and the number of cells permeating through the filter membrane decreased following transfection with siRNA. The inhibition of PLK1 caused a significant decrease in CD44v6, matrix metalloproteinase (MMP)-2 and MMP-9 (0.36±0.08, 0.12±0.03, 0.25±0.06, respectively) compared to the non-transfected group (1.15±0.18, 1.21±0.20, 1.25±0.21, respectively; P<0.01). In conclusion, the expression of PLK1 was found to be increased in anaplastic thyroid carcinoma and was correlated with clinical stage, lymph node metastasis and prognosis. Additionaly, PLK1 siRNA was found to inhibit the invasion of anaplastic thyroid carcinoma cells. Therefore, CD44v6, MMP-2 and MMP-9 are likely to be involved in the regulation of cell invasion induced by PLK1.
本研究旨在观察polo样激酶1(PLK1)基因在间变性甲状腺癌细胞侵袭中的调控作用,并探讨其机制。采用免疫组织化学染色法检测36例间变性甲状腺癌患者中PLK1蛋白的表达。设计、合成针对PLK1的小干扰RNA(siRNA)并转染至ARO细胞。采用软琼脂集落形成试验和Transwell小室试验检测PLK1 siRNA对细胞侵袭的影响。使用蛋白质免疫印迹分析检测相应蛋白。间变性甲状腺癌样本中PLK1的表达(67.5±10.6%)显著高于癌旁样本(0.65%±0.12%;P<0.01)。PLK1的表达与间变性甲状腺癌的临床分期、淋巴结转移及预后相关。随着siRNA水平升高,细胞克隆数呈剂量依赖性减少,转染siRNA后穿过滤膜的细胞数减少。与未转染组相比(分别为1.15±0.18、1.21±0.20、1.25±0.21),抑制PLK1导致CD44v6、基质金属蛋白酶(MMP)-2和MMP-9显著降低(分别为0.36±0.08、0.12±0.03、0.25±0.06;P<(此处疑似原文有误,应为P<0.01))。总之,发现PLK1在间变性甲状腺癌中表达增加,且与临床分期、淋巴结转移及预后相关。此外,发现PLK1 siRNA可抑制间变性甲状腺癌细胞的侵袭。因此,CD44v6、MMP-2和MMP-9可能参与PLK1诱导的细胞侵袭调控。