Loss of primary cilia promotes migration of TPC-1 in papillary thyroid cancers
马承旭 1马小妮 1马丽华 1傅松波 1朱鋐达
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作者信息
1. 兰州大学第一临床医学院，兰州大学第一医院内分泌科，兰州 730000
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摘要
目的 探讨miR-182-5p调控初级纤毛的缺失对甲状腺乳头状癌(PTC)细胞系TPC-1迁移的影响。 方法 收集10例兰州大学第一医院PTC组织和癌旁组织，qPCR检测癌旁组织、PTC组织和TPC-1细胞中miR-182-5p的表达量，免疫荧光检测初级纤毛的形成；过表达miR-182-5p，qPCR和Western blot检测TPC-1细胞中PI3K的表达量，Transwell实验检测TPC-1细胞迁移数量，免疫荧光检测初级纤毛的形成；siRNA-IFT88干扰TPC-1细胞，Transwell实验检测TPC-1细胞迁移数量，免疫荧光检测初级纤毛的形成，吉姆萨染色检测形态学变化，Western blot检测E-cadherin和N-cadherin的蛋白表达量。 结果 与癌旁组织相比，PTC组织中初级纤形成受损；miR-182-5p表达量显著上调。与Nthy-ori3-1相比，TPC-1细胞中初级纤毛的频率减少；miR-182-5p表达量显著上调(P＝0.001)；miR-182-5p mimic治疗后的TPC-1细胞中PI3K的表达量降低，迁移数量增加(P＝0.001)，初级纤毛频率降低为27%(P＝0.002)；siRNA-IFT88治疗后的TPC-1细胞表面初级纤毛变短变细，频率减少(P＝0.001)，迁移数量增加(P<0.001)。 结论 miR-182-5p通过PI3K通路调控初级纤毛的缺失有助于TPC-1细胞的迁移，初级纤毛的丢失对PTC预后具有不利的影响。 Objective To investigate the role of miR-182-5p regulated primary cilia loss on migration of TPC-1 in papillary thyroid cancers(PTC). Methods Ten cases of PTC and adjacent tissues were collected from the First Hospital of Lanzhou University, The expression of miR-182-5p in PTC tissue and TPC-1 cells was detected by qPCR, and the frequency of primary cilia was detected by immunofluorescence Overexpressing miR-182-5p, the migrated number of TPC-1 was detected by Transwell assay Interfering TPC-1 with siRNA-IFT88, the migrated number of TPC-1 and the frequency of primary cilia were detected, respectively. Results Compared with control, the expression of miR-182-5p was significantly upregulated in PTC and TPC-1, the frequency of primary cilia in PTC and TPC-1 was downregulated. Overexpressing miR-182-5p increased the migrated number of TPC-1 cell and reduced the number of TPC-1 cell migration(27%, P=0.002) After siRNA-IFT88 treatment, primary cilia in TPC-1 became shorter and thinner, with a decrease in frequency(P=0.001), the migrated number of TPC-1 cell increased, and TPC-1 cell showed smaller nuclei and fewer microvilli. Conclusion The regulation of primary cilia loss by miR-182-5p through the PI3K pathway contributes to the migration of TPC-1 cells. The loss of primary cilia has an adverse impact on the prognosis of PTC.
Abstract
Objective To investigate the role of miR-182-5p regulated primary cilia loss on migration of TPC-1 in papillary thyroid cancers(PTC). Methods Ten cases of PTC and adjacent tissues were collected from the First Hospital of Lanzhou University, The expression of miR-182-5p in PTC tissue and TPC-1 cells was detected by qPCR, and the frequency of primary cilia was detected by immunofluorescence Overexpressing miR-182-5p, the migrated number of TPC-1 was detected by Transwell assay Interfering TPC-1 with siRNA-IFT88, the migrated number of TPC-1 and the frequency of primary cilia were detected, respectively. Results Compared with control, the expression of miR-182-5p was significantly upregulated in PTC and TPC-1, the frequency of primary cilia in PTC and TPC-1 was downregulated. Overexpressing miR-182-5p increased the migrated number of TPC-1 cell and reduced the number of TPC-1 cell migration(27%, P=0.002) After siRNA-IFT88 treatment, primary cilia in TPC-1 became shorter and thinner, with a decrease in frequency(P=0.001), the migrated number of TPC-1 cell increased, and TPC-1 cell showed smaller nuclei and fewer microvilli. Conclusion The regulation of primary cilia loss by miR-182-5p through the PI3K pathway contributes to the migration of TPC-1 cells. The loss of primary cilia has an adverse impact on the prognosis of PTC.
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