膜联蛋白A1对甲状腺乳头状癌细胞增殖、迁移及侵袭能力的影响  

Effect of annexin A1 on the proliferation,migration and invasion of papillary thyroid carcinoma

作　　者：陈立 田志刚 贾兰宁 杨洋 赵科[1,2] 王义增 何向辉[1] Chen Li;Tian Zhigang;Jia Lanning;Yang Yang;Zhao Ke;Wang Yizeng;He Xianghui(Department of General Surgery,General Hospital of Tianjin Medical University,Tianjin 300052,China;Tianjin Institute of General Surgery,Tianjin 300052,China;Tianjin Union Medical Center,Tianjin 300191,China)

机构地区：[1]天津医科大学总医院普通外科,天津300052 [2]天津市普通外科研究所,天津300052 [3]天津市人民医院,天津300191

出　　处：《中华内分泌外科杂志》2022年第1期23-27,共5页Chinese Journal of Endocrine Surgery

基　　金：国家自然科学基金(81672641)。

摘　　要：目的通过短发夹RNA(shRNA)干扰膜联蛋白A1(AnnexinA1,ANXA1)在甲状腺乳头状癌(papillary thyroid carcinoma,PTC)细胞系中的表达,探讨ANXA1对PTC细胞的生物行为学影响。方法设计使用具有专一高效性的shRNA在TPC-1和BCPAP细胞系中特异性干扰ANXA1的表达,分别对TPC-1及BCPAP细胞系进行转染,包括特异性ANXA1干扰及阴性对照病毒转染,分为shANXA1组和阴性对照病毒组,然后通过半定量逆转录PCR(Q-PCR)与蛋白免疫印迹法(Western Blot)验证基因表达,以shANXA1组为实验组,未转染病毒组及阴性对照病毒组为对照组,比较三组细胞ANXA1的表达水平,验证shRNA干扰效率,再通过划痕、CCK8、Transwell侵袭实验探究ANXA1敲减后对TPC-1和BCPAP细胞系增殖、迁移及侵袭能力的影响。使用独立样本t检验比较两组间均数,多组间比较采用单因素方差分析,P<0.05差异具有统计学意义。结果shRNA可高效沉默ANXA1在PTC细胞系中转录和翻译水平上的表达,通过慢病毒转染成功后的TPC-1和BCPAP和阴性对照组细胞相比,细胞增殖(BCPAP,24 h:F=25.15,P<0.001;48 h:F=6.44,P<0.001;48 h:F=46.94,P<0.001;TPC-1,24 h:F=207.50,P<0.001;48 h:F=202.45,P<0.001;48 h:F=55.89,P<0.001),迁移(BCPAP,F=12511.10,P<0.001;TPC-1,F=3966.10,P<0.001)及侵袭能力(BCPAP:F=94.65,P<0.001;TPC-1:F=681.74,P<0.001)明显下降。结论shRNA敲减ANXA1基因后,TPC-1及BCPAP细胞系增殖、迁移及侵袭能力显著下降,表明沉默该基因可降低肿瘤的侵袭性,并初步揭示ANXA1可能是PTC生物治疗中一个重要的潜在靶点。Objective To investigate the effect of ANXA on biological behavior of papillary thyroid carcinoma(PTC)cells by interfering with the expression of annexin A1(ANXA1)in PTC cell lines by short hairpin RNA(shRNA).Methods The shRNA with specific and high efficiency was designed to specifically interfere with the expression of ANXA1 in TPC-1 and BCPAP cell lines,and transfect the TPC-1 and BCPAP cell lines respectively,including specific ANXA1 interference and negative control virus transfection,and they were divided into shANXA1 group and negative control virus group.Semi-quantitative reverse transcription PCR(Q-PCR)and Western Blot were employed to verify gene expression.The shANXA1 group was used as the experimental group,the untransfected virus group and the negative control virus group were set as the control groups.The expression levels of ANXA1 in the three groups were compared and the shRNA interference efficiency was verified.The effects of ANXA1 knockdown on the proliferation,migration and invasion of TPC-1 and BCPAP cell lines were investigated by scratch,CCK8 and Transwell invasion experiments.Independent sample t test was used to compare the means between the two groups,and one-way analysis of variance was employed to compare multiple groups,with P<0.05 as statistically significant.Results shRNA could efficiently silence the expression of ANXA1 at the transcription and translation level in PTC cell lines.Compared with the negative control cells,the cells proliferated after successful lentiviral transfection of TPC-1 and BCPAP(BCPAP,24h:F=25.15,P<0.001;48h:F=6.44,P<0.001;48h:F=46.94,P<0.001;TPC-1,24h:F=207.50,P<0.001;48h:F=202.45,P<0.001;48h:F=55.89,P<0.001),its migration(BCPAP,F=12511.10,P<0.001;TPC-1,F=3966.10,P<0.001)and invasion ability(BC-PAP:F=94.65,P<0.001;TPC-1:F=681.74,P<0.001)significantly decreased.Conclusion After shRNA knock-down of ANXA1 gene,the proliferation,migration and invasion ability of TPC-1 and BCPAP cell lines decreased significantly,indicating that silencing this gene can reduce tum

关 键 词：短发夹RNA 膜联蛋白 甲状腺乳头状癌 细胞功能实验 

分 类 号：R653[医药卫生—外科学] R736.1[医药卫生—临床医学]