雷公藤红素通过逆转EMT抑制SKOV-3细胞的侵袭转移

Celastrol inhibits migration and invasion of SKOV-3 cells by reversing EMT

目的:探讨雷公藤红素对卵巢癌SKOV-3细胞侵袭转移的抑制作用及机制。方法:应用MTT、Transwell小室实验、划痕修复实验测定细胞的增殖、侵袭、转移能力; Western blot法检测各组相关蛋白的表达。结果:MTT显示雷公藤红素抑制SKOV-3细胞增殖,呈时间和剂量依赖性; Transwell小室实验显示空白组、TGF-β1组、TGF-β1+雷公藤红素(0. 24、0. 36、0. 48mg/L)组、TGF-β1+SIS3 HCI组(n=3)侵袭细胞数分别为262±5、345±7、305±4、147±6、83±5、189±8;划痕修复实验显示各组细胞12h迁移率为(41±3)%、(56±5)%、(33±5)%、(24±2)%、(13±3)%、(31±4)%,24h迁移率为(54±6)%、(67±5)%、(42±1)%、(37±3)%、(28±7)%、(40±5)%,与模型组比较有显著差异; Western blot显示,与空白组相比,TGF-β1组E-Cadherin表达下调,Vimentin、Fib、Snail表达上调;与TGF-β1组相比,TGF-β1+雷公藤红素(0. 24、0. 36、0. 48mg/L)组和TGF-β1+SIS3 HCI组E-Cadherin表达上调,Vimentin、Fib、Snail表达下调,同时p-Smad2/3的表达亦明显下调,Smad2/3表达显著上调。结论:雷公藤红素抑制SKOV-3细胞的侵袭转移,机制可能是通过阻断TGF-β/Smad信号通路,逆转细胞EMT的发生。

Objective： To investigate the inhibitory effects of celastrol on migration and invasion of ovarian cancer SKOV-3 cells and its mechanism.Methods： MTT assay,transwell chamber experiment and scratch test were used to measure proliferation,migration and invasion. And western blot was applied to detect the expressions of protein. Results： MTT showed that celastrol inhibited the proliferation of SKOV-3 cells in a time and dose dependent manner. Transwell chamber experiment showed the number of invasion cells of the blank group,TGF-β1 group,TGF-β1 ＋ celastrol（0. 24,0. 36,0. 48 mg/L） group and TGF-β1 ＋ SIS3 HCI group were 262 ± 5,345 ± 7,305 ± 4,147 ± 6,83 ± 5,189 ±8; the scratch test showed the migration rates at 12 h were（41 ± 2. 8） %,（56 ± 4. 6） %,（33 ± 5. 1） %,（24 ± 1. 6） %,（13 ± 3. 2） %,（31± 4. 2） %; at 24 h,they were（54 ± 6. 1） %,（67 ± 5. 3） %,（42 ± 1. 2） %,（37 ± 2. 5） %,（28 ± 7. 1） %,（40 ± 4. 6） %; the difference was statistically significant（P 〈 0. 05）. Western blot results showed that compared with the blank group,the expression of E-candherin in TGF-β1 group was down-regulated and the expression of Vimentin,Fib and Snail was up-regulated. Compared with TGF-β1 group,in TGF-β1 ＋celastrol（0. 24、0. 36、0. 48 mg/L） group and TGF-β1 ＋ SIS3 HCI group,the expression of E-Candherin was up-regulated,and the expression of Vimentin,Fib and Snail was down-regulated. Meanwhile,the expression of p-Smad2/3 was also down-regulated. On the contrary Smad2/3 was down-regulated（P 〈 0. 05）. Conclusion： Celastrol can inhibit migration and invasion of ovarian cancer SKOV-3 cells,which may reverse the occurrence of EMT in SKOV-3 by inhibiting TGF-β/Smad signaling pathway.