Objective: To investigate the effects of bosutinib on the malignant behavior of thyroid papillary carcinoma B-CPAP cells and its possible mechanisms. Methods: Thyroid papillary carcinoma B-CPAP cells were cultured in vitro with a concentration gradient of(1、2、3、4 and 5 μmol/L)bosutinib intervened for 24 hours, DMSO was used as the control group. Five parallel compound holes were set in each group. Cell counting kit (CCK-8 method) method was used to detect cell proliferation. Transwell assay and cell wound healing assay were used to detect cell invasion and migration. TUNEL staining assay and flow cytometry were used to detect cell apoptosis. Western blot was used to detect the expressions of autophagic proteins (Beclin-1, LC3, p62) and signal pathway proteins (SIK2, p-mTOR, mTOR, p-ULK1, ULK1). Results: Compared with the control group, the cell proliferation activity, migration ability and invasion ability were decreased (P＜0.01), while the cell apoptosis rate was increased (P＜0.01) in the bosutinib concentration groups of 2, 3, 4 and 5 μmol/L . In the concentration groups of 4 and 5 μmol/L, the expression of Beclin-1 (P＜0.05), LC3- Ⅱ/LC3- Ⅰ (P＜0.05), SIK2 (P＜0.01) and p-ULK1 (P＜0.01) protein was decreased, while the expression of p62 (P＜ 0.05) and p-mTOR (P＜0.01) protein was increased. Conclusion: Bosutinib may inhibit the autophagy of thyroid papillary carcinoma cells through SIK2-mTOR-ULK1 signaling pathway to inhibit their proliferation, invasion and migration and promote apoptosis, thereby weakening their malignant behavior.
目的: 探讨博舒替尼对甲状腺乳头状癌B-CPAP细胞恶性行为的影响及其可能的作用机制。 方法: 体外培养甲状腺乳头状癌B-CPAP细胞系,加入浓度梯度(1、2、3、4和5 μmol/L)博舒替尼干预24 h,用DMSO作为对照组,每组设5个平行复孔。采用CCK-8法检测细胞增殖能力,Transwell实验和划痕修复实验检测细胞侵袭和迁移能力,TUNEL染色实验和流式细胞术检测细胞凋亡情况,蛋白质印迹法检测自噬蛋白(Beclin-1、LC3、p62)表达情况和信号通路蛋白(SIK2,p-mTOR、mTOR 、p-ULK1、ULK1)表达情况。 结果: 与对照组比较,博舒替尼2、3、4和5 μmol/L浓度组细胞增殖活性、迁移能力和侵袭能力均降低(P＜0.01)而细胞凋亡率均增加(P＜0.01),博舒替尼4和5μmol/L浓度组Beclin-1(P＜0.05)、LC3-Ⅱ/LC3-Ⅰ(P＜0.05)、SIK2(P＜0.01)和p-ULK1(P＜0.01)蛋白表达量均减少而p62(P＜0.05)和p-mTOR(P＜0.01)蛋白表达量增加。结论: 博舒替尼可能通过 SIK2-mTOR-ULK1通路抑制甲状腺乳头状癌细胞自噬,来抑制其增殖、侵袭和迁移能力并促进凋亡,从而减弱其恶性行为。.
Keywords: Bosutinib; autophagy; cell culture; malignant behavior; thyroid papillary carcinoma.