摘要
目的尽管TSH被认为和甲状腺癌的发病率和预后密切相关,但是具体潜在机制还没有被阐明。本研究拟探讨乳头状甲状腺癌中TSH是否能调控Cpt1c表达,同时研究TSH对Cpt1c促进乳头状甲状腺癌细胞增殖的影响。方法首先收集甲状腺癌及其配对的癌旁正常组织标本,利用qRT-PCR法检测Cpt1c基因在35对乳头状甲状腺癌标本及其配对的癌旁正常组织的表达情况;其次分别在浓度梯度和时间梯度上检测TSH对甲状腺癌细胞株B-CPAP中Cpt1c表达的影响;最后构建Cpt1c干扰RNA,转染B-CPAP细胞株,利用免疫印迹法检测和qRT-PCR方法对肿瘤细胞转染干扰RNA效率进行鉴定。鉴定完毕后,利用CCK-8和FACS细胞仪分别检测TSH刺激下的细胞增殖和细胞周期。结果 Cpt1c在乳头状甲状腺癌组织中高表达于正常癌旁组织;TSH可促进乳头状甲状腺癌细胞株B-CPAP中的Cpt1c表达,并且呈浓度依赖性,随着浓度的增高,Cpt1c表达亦增加;Cpt1c干扰可通过S期停滞影响甲状腺癌细胞增殖,同时可通过促进Cpt1c表达引起细胞进入G2期从而引起更强的增殖能力。结论 TSH通过调节Cpt1c表达可以促进乳头状甲状腺癌细胞增殖,Cpt1c基因将来可能成为乳头状甲状腺癌基因治疗的新靶点。
Objective Though TSH is closely related to the incidence and prognosis of thyroid carcinoma,the underlying mechanisms have not been elucidated.This study was to investigate whether thyroid stimulating hormone in papillary thyroid carcinoma can regulate the expression of Cpt1c and to investigate the effect of Cpt1c on TSH promoting the proliferation of papillary thyroid carcinoma cells.Methods Firstly,the expression of Cpt1c was detected by qRT-PCR in35papillary thyroid carcinoma specimens and their adjacent normal tissues from papillary thyroid carcinoma patients.Secondly,the effects of TSH on the expression the Cpt1c from B-CPAP was detected at time and concentration gradient respectively.Finally,Cpt1c siRNA was constructed and transfected into B-CPAP.Western blot and qRT-PCR were used to identify the siRNA transfection efficiency of tumor cells.After identification,CCK-8and FACS were used to detect cell proliferation and cell cycle with stimulation of TSH.Results Cpt1c is highly expressed in human papillary thyroid cancer compared with paired normal tissues.TSH could promote the expression of Cpt1cin a dose-dependent manner.Cpt1c could affect the proliferation of thyroid cancer cells by S phase stagnation and the expression ofCpt1c in papillary thyroid carcinoma is higher than that in adjacent tissues.At the same time,TSH could promote Cpt1c expressioncaused by cells into the G2phase to increase proliferation.Conclusions TSH could promote the proliferation of papillary thyroid carcinoma cells by regulating the expression of Cpt1c.Cpt1c gene may be a new target for the treatment of papillary thyroid cancer in the future.
作者
王瑞
逄志骏
周鑫宇
程令涛
巩博深
何晓波
缪明永
姚真真
苏东玮
WANG Rui;PANG Zhi-jun;ZHOU Xin-yu;CHENG Ling-tao;GONG Bo-shen;HE Xiao-bo;MIAO Ming-yong;YAO Zhen-zhen;SU Dong-wei(Department of Biochemistry and Molecular Biology, Second Military Medical University, Shanghai 200433, China;Department of General Surgery (Surgical Breast and Thyroid Section), Changhai Hospital, Second Military Medical University, Shanghai 200433,China)
出处
《肿瘤代谢与营养电子杂志》
2017年第1期92-96,共5页
Electronic Journal of Metabolism and Nutrition of Cancer
基金
国家自然科学基金(81372862)
上海市卫生局(2012158)
