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CD44+CD24-～low,p63的表达与甲状腺乳头状癌细胞转移侵袭能力的研究
The expression of CD44+/CD24-～low and p63 are associated with the lymph node metastases of papillary thyroid carcinoma

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DOI:
作者:
陈晓军，郭晶晶，李天一，廖芝伟，董建，周远航，黄琦
Xiaojun chen, Jingjing Guo, Tianyi li, Zhiwei Liao, Jian Dong, Yuanhang Zhou, Qi Huang
作者单位:
上海市宝山区仁和医院
Renhe Hospital, Baoshan District, Shanghai
关键词:
CD44+/CD24-～low，p63，甲状腺乳头状癌，淋巴结转移
CD44+/CD24-～low,p63,papillary thyroid carcinoma(PTC),lymph node metastases(LNM)
摘要:
目的：探讨CD44+/CD24-～low亚群比例以及p63蛋白表达水平对甲状腺乳头状癌细胞体外转移侵袭能力的影响。 方法：选取TPC-1、GLAG-66、BCPAP和NPA四株甲状腺乳头状癌细胞株进行体外实验，流式细胞术检测CD44+/CD24-～low亚群比例； Western blot方法检测p63蛋白的分布情况； Transwell侵袭实验检测细胞的转移侵袭能力；CCK-8法观察多柔比星对不同侵袭能力肿瘤细胞生长的抑制作用；qRT-PCR 检测TAp63和ΔNp63 mRNA表达情况。 结果：CD44+/CD24-～low亚群比例在TPC-1、BCPAP、GLAG-66、NPA和正常甲状腺细胞株Nthy-ori 3-1中分别为34.59%±1.88%、0.28%±0.015%、44.15%±1.42%、 56.96%±1.94%和0.25±0.012%，存在显著差异（P<0.000），BCPAP和Nthy-ori 3-1的CD44+/CD24-～low亚群比例无显著差异（P=0.826）。p63蛋白在各甲状腺乳头状癌细胞株中均有表达，但在Nthy-ori 3-1细胞几乎无表达。亚群比例高的肿瘤细胞同时表现为p63蛋白高表达，其迁移、侵袭能力和抗化疗药物能力显著高于对照组和低亚群比例组(P<0.001)。ΔNp63 mRNA在TPC-1、BCPAP、 GLAG-66、 NPA和正常甲状腺细胞株Nthy-ori 3-1中表达水平分别为0.00057±0.00016、5.7E-05±6.8E-06、0.0008±8.5E-05和0.0013±9.1E-05，差异有明显的统计学意义（P<0.05）。 结论：甲状腺乳头状癌细胞株的转移、侵袭能力与CD44+/CD24-～low亚群比例和p63表达相关，CD44和p63对于未来靶向治疗的靶点选择具有一定参考意义。
Objective To explore the relationship between CD44+/CD24-～lowand p63 protein levels and the ability of invasion and metastases (LNM) in papillary thyroid carcinoma (PTC) cell lines. Methods Four groups of papillary thyroid carcinoma cells(included TPC-1, BCPAP, GLAG-66, NPA) were treated in vitro laboratory experiments.Flow Cytometry was performed to evaluate the proportion of the subpopulation of CD44+/CD24-～low. Western blot were used to detect the p63 protein expression in a normal thyroid cell line and 4 PTC cell lines including TPC-1, BCPAP, GLAG-66 and NPA. 24-well Transwell test were used to evaluate the ability of cell migration. CCK-8 assays were performed to test the resistance against chemotherapy drug among PTC cell lines. Meanwhile,qRT-PCR was also conducted to detect the TAp63 and ΔNp63 messenger RNA (mRNA) in PTC cells. Result The proportion of CD44+/CD24-～low in papillary thyroid carcinoma cells(TPC-1, BCPAP, GLAG-66, NPA) and normal thyroid cells(Nthy-ori 3-1) were 34.59%±1.88%, 0.28%±0.015%, 44.15%±1.42%, 56.96%±1.94%and 0.25±0.012%，which were significant differences（P<0.000）.However,there were no statistical differences between groups of BCPAP and Nthy-ori 3-1（P=0.826）. Expression of p63 protein in the papillary thyroid carcinoma cell line in both, but almost no expression of 3-1 cells in Nthy-ori.The PTC cell lines with a higher proportion of CD44+/CD24-～low tended to show a high expression of p63, and have significantly strong ability of invasion, migration and resistance of chemotherapy drug compared with control team and the lower ones(P<0.001).The expression value of ΔNp63 messenger RNA (mRNA) in TPC-1, BCPAP, GLAG-66, NPA and normal thyroid cells (Nthy-ori 3-1 ) were: 0.00057 + 0.00016, 5.7E-05 + 6.8E-06, 0.0008 + 8.5E-05, 0.0013 + 9.1E-05, and there were statistical significance difference between them (P <0.05). Conclusion PTC cells’ ability of migration and invasion may be related to the proportion of CD44+/CD24-～low and expression of p63 in different cell lines. These two markers might be new targets for molecular targeted therapy in the future.

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