摘要
目的探讨甲状腺素(T_(4))能否通过整合素α_(v)β_(3)影响分化型甲状腺癌(DTC)细胞增殖、转移潜能及其分子机制。方法体外培养甲状腺乳头状癌TPC-1、K1及甲状腺滤泡状癌(FTC)133细胞株,采用免疫荧光和流式细胞术分析细胞表面整合素α_(v)β_(3)的表达水平。给予T_(4)、四碘甲状腺乙酸(Tetrac)、精氨酸-甘氨酸-天冬氨酸(RGD)多肽单独或联合处理DTC细胞后,应用细胞计数试剂盒-8(CCK-8)法及Transwell小室迁移和侵袭实验检测细胞增殖和转移潜能的变化。通过小干扰RNA(siRNA)转染模型验证整合素α_(v)或β_(3)亚基沉默能否逆转T_(4)对DTC细胞的作用。利用Western blot法检测下游信号蛋白磷酸化细胞外信号调节激酶(p-ERK)1/2、总细胞外信号调节激酶(ERK)1/2的表达水平。应用丝裂原活化蛋白激酶的激酶(MEK)1/2的抑制剂GSK1120212阻断ERK1/2磷酸化,检测其对T_(4)处理细胞增殖、迁移和侵袭能力的影响。多组间比较采用单因素方差分析,组间两两比较采用Tukey法。结果TPC-1、K1和FTC133细胞整合素α_(v)β_(3)表达均呈阳性,相对平均荧光强度(MFI)分别为61.93±18.61、16.89±2.43和32.36±0.83,阳性细胞百分比分别为(94.38±1.30)%、(74.11±3.87)%和(50.67±1.78)%(F值:13.36和217.30,P值:0.006和<0.001)。T_(4)组TPC-1、K1和FTC133细胞的增殖、迁移和侵袭能力较对照组明显增强(96 h,F值:62.67~297.50,q值:13.15~20.73,均P<0.001);T_(4)+Tetrac和T_(4)+RGD多肽处理组DTC细胞的增殖、迁移和侵袭能力较T_(4)组明显减低(96 h,q值:8.61~17.54,均P<0.001)。利用siRNA敲减整合素α_(v)或β_(3)亚基可明显抑制T_(4)对3种DTC细胞的促增殖、迁移和侵袭作用(72 h,F值:7.75~70.98,q值:4.77~15.21,均P<0.05)。Western blot结果显示,T_(4)处理导致DTC细胞ERK1/2磷酸化水平明显升高,且T_(4)诱导的ERK1/2信号通路激活可被Tetrac、RGD多肽、整合素α_(v)或β_(3)亚基敲减所阻断。GSK1120212可以明显逆转T_(4)�
Objective To explore whether thyroxine(T_(4))could promote differentiated thyroid cancer(DTC)progression by binding to integrinα_(v)β_(3)in vitro and its downstream mechanism.Methods Papillary thyroid cancer cell lines TPC-1,K1 and follicular thyroid cancer(FTC)cell line FTC133 were cultured in vitro,and the expressions of integrinα_(v)β_(3) in those 3 DTC cell lines were determined with immunofluorescence and flow cytometry analysis.After the treatment of T_(4),tetraiodo thyroacetic acid(Tetrac)and Arg-Gly-Asp(RGD)peptide alone or in combination,the proliferation and metastatic potential of DTC cell lines were detected by cell counting kit-8(CCK-8),Transwell migration and invasion assays.The small interfering RNA(siRNA)transfection was used to verify whether integrinα_(v) orβ_(3) subunit knockdown could reverse the effect of T_(4) on DTC cells.The expression levels of downstream signaling proteins phosphorylated extracellular signal-regulated kinase(p-ERK)1/2 and total extracellular signal-regulated kinase(ERK)1/2 were detected by Western blot.The effects of mitogen-activated protein kinase kinase(MEK)1/2 inhibitor(GSK1120212)on the proliferation,migration and invasion of T_(4)-treated cells were detected.One-way analysis of variance and Tukey test were used for data analysis.Results The integrinα_(v)β_(3) expressions in TPC-1,K1 and FTC133 cells were all positive,with the relative mean fluorescence intensity(MFI)of 61.93±18.61,16.89±2.43 and 32.36±0.83,and the percentages of positive cells of(94.38±1.30)%,(74.11±3.87)%and(50.67±1.78)%,respectively(F values:13.36 and 217.30,P=0.006 and P<0.001).Compared with control group,the proliferation,migration and invasion in the three DTC cell lines treated with T_(4) were significantly enhanced(96 h,F values:62.67-297.50,q values:13.15-20.73,all P<0.001).T_(4)-induced cell proliferation,migration and invasion were markedly reversed by Tetrac or RGD(96 h,q values:8.61-17.54,all P<0.001).T_(4)-induced cell proliferation,migration and invasion were also signif
作者
梁一倩
贾茜
王源波
李慧婕
杨艺员
张月敏
许惠
杨爱民
高蕊
Liang Yiqian;Jia Xi;Wang Yuanbo;Li Huijie;Yang Yiyuan;Zhang Yuemin;Xu Hui;Yang Aimin;Gao Rui(Department of Nuclear Medicine,the First Affiliated Hospital of Xi′an Jiaotong University,Xi′an 710061,China)
出处
《中华核医学与分子影像杂志》
CAS
CSCD
北大核心
2023年第8期452-458,共7页
Chinese Journal of Nuclear Medicine and Molecular Imaging
基金
国家自然科学基金(81902322)
陕西省自然科学基础研究计划(2020JQ-526)。
