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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。
Purpose Although TSH is thought to be closely related to the incidence and prognosis of thyroid cancer, the exact underlying mechanisms have not yet been elucidated. This study was to investigate whether TSH could regulate the expression of Cpt1c in papillary thyroid carcinoma and to study the effect of TSH on the proliferation of papillary thyroid carcinoma cells by Cpt1c. Methods Thirty-five specimens of papillary thyroid carcinoma and normal paracancer tissues were collected by qRT-PCR method. The expression of Cpt1c gene was detected by qRT-PCR. The effect of TSH on the expression of Cpt1c in thyroid cancer cell line B-CPAP was detected by the gradient method. Finally, the Cpt1c interfering RNA was constructed and transfected into B-CPAP cell line. The efficiency of transfected tumor cells transfected with interfering RNA was detected by Western blotting and qRT-PCR Identification. After the identification, CCK-8 and FACS were used to detect the proliferation and cell cycle of TSH-stimulated cells respectively. Results Cpt1c was highly expressed in normal papillary thyroid carcinoma tissues. TSH promoted Cpt1c expression in B-CPAP cells in a dose-dependent manner. Cpt1c expression was also increased with increasing concentration of Cpt1c. Cpt1c interference can affect the proliferation of thyroid cancer cells through the stagnation of S phase, and at the same time, it can lead to stronger proliferation through promoting the expression of Cpt1c to cause cells to enter G2 phase. Conclusion TSH can promote the proliferation of papillary thyroid carcinoma cells by regulating the expression of Cpt1c, and Cpt1c may become a new target of gene therapy of papillary thyroid carcinoma in the future.