Yuan Yanyan, Lan Yuanzao, Cui Yi, Hou Minhua, Tang Jinyong, Wen Haojie
Department of Otolaryngology-Head and Neck Surgery，Chenzhou No. 1 People's Hospital，Chenzhou，423000，China.
The First School of Clinical Medicine，Southern Medical University.
Lin Chuang Er Bi Yan Hou Tou Jing Wai Ke Za Zhi. 2024 Mar;38(3):188-194;199. doi: 10.13201/j.issn.2096-7993.2024.03.002.
To investigate the effects and molecular mechanisms of abietic acid in the cell proliferation, invasion and migration of cisplatin-resistant nasopharyngeal carcinoma cells. ①Cisplatin-resistant C666/DDP cell line was constructed by increasing drug concentration method. ②The effects of abietic acid on proliferation, invasion and migration of C666/DDP cells were investigated by CCK-8 method, reactive oxygen species（ROS） and mitochondrial membrane potential（MMP） level assay and subcutaneous tumorigenesis assay in nude mice to detect the effects of abietic acid on proliferation and apoptosis of C666/DDP cells in vitro and in vivo. The effect of abietic acid on the proliferation and apoptosis of C666/DDP cells in vitro and in vivo was measured by Transwell assay. ③Western blot and IHC method to detect the expression of PI3K/AKT/mTOR pathway related proteins. ①The IC50 of cisplatin cytotoxicity to C666-1 was about 25 μmol/L. RI=25 μmol/L /4 μmol/L=6.25, resistance was obtained, and the C666-1-DDP resistant strain was successfully constructed. ②Abietic acid promoted apoptosis and inhibited proliferation of C666/DDP cells, and showed G2/M phase block; transwell showed that abietic acid inhibited C666/DDP cell migration and invasion, increased ROS level of C666/DDP cells and decreased MMP. Transwell showed that abietic acid inhibited the migration and invasion ability of C666/DDP cells, increased the ROS level of C666/DDP cells and decreased MMP. ③Animal experiments showed that abietic acid inhibited the proliferation of cisplatin-resistant nasopharyngeal carcinoma in vivo in a concentration gradient and suppressed the expression of PI3K/AKT/mTOR signaling pathway-related proteins. Abietic acid inhibits proliferation, invasion and migration of cisplatin-resistant nasopharyngeal carcinoma cells by a mechanism related to inhibition of PI3K/AKT/mTOR signaling pathway.
探讨枞酸对顺铂耐药鼻咽癌细胞增殖、侵袭和迁移的影响及其分子机制。①采用逐步增加药物浓度的方法构建顺铂耐药C666/DDP细胞系。②通过CCK-8法、活性氧（ROS）和线粒体膜电位（MMP）水平检测以及裸鼠皮下成瘤实验，研究枞酸对C666/DDP细胞增殖、侵袭和迁移的影响，以检测枞酸在体外和体内对C666/DDP细胞增殖和凋亡的作用。通过Transwell实验检测枞酸在体外和体内对C666/DDP细胞增殖和凋亡的影响。③采用蛋白质免疫印迹法（Western blot）和免疫组化法（IHC）检测PI3K/AKT/mTOR信号通路相关蛋白的表达。①顺铂对C666-1细胞的半数抑制浓度（IC50）约为25 μmol/L。耐药指数（RI）=25 μmol/L /4 μmol/L = 6.25，成功获得耐药性，构建了C666-1-DDP耐药株。②枞酸促进C666/DDP细胞凋亡并抑制其增殖，呈现G2/M期阻滞；Transwell实验表明枞酸抑制C666/DDP细胞迁移和侵袭，增加C666/DDP细胞的ROS水平并降低MMP。③动物实验表明，枞酸在体内以浓度梯度抑制顺铂耐药鼻咽癌的增殖，并抑制PI3K/AKT/mTOR信号通路相关蛋白的表达。枞酸通过抑制PI3K/AKT/mTOR信号通路相关机制抑制顺铂耐药鼻咽癌细胞的增殖、侵袭和迁移。
