姜黄素通过miR-206/BDNF通路抑制宫颈癌HeLa细胞的侵袭和迁移

Curcumin inhibits invasion and migration of cervical cancer HeLa cells via miR-206/BDNF pathway

目的探讨姜黄素对宫颈癌HeLa细胞侵袭转移的抑制作用及其调控机制。方法姜黄素分别以50μmol/L处理0、24、36、48、72 h干预宫颈癌HeLa细胞后,采用MTT法观察其对HeLa细胞活力的影响。应用Transwell小室检测姜黄素对HeLa细胞侵袭转移能力的抑制作用,免疫印迹实验检测其对金属基质蛋白酶-2、9(MMP-2和MMP-9)和脑源性神经营养因子(BDNF)表达水平的影响。通过RT-qPCR检测姜黄素对miR-206和BDNF表达的影响及双荧光素酶实验检测miR-206对BDNF的调控关系。结果姜黄素可抑制HeLa细胞的活力,且呈现时间依赖性;姜黄素可显著降低HeLa细胞的侵袭和迁移能力并能够降低MMP-2和MMP-9的表达水平,同时激活miR-206的表达;双荧光素酶结果显示,miR-206通过靶向降解BDNF来抑制HeLa细胞的侵袭和迁移能力。结论姜黄素通过激活miR-206/BDNF通路抑制宫颈癌HeLa细胞的侵袭和迁移。

Objective To investigate the effect of curcumin against invasion and metastasis of cervical cancer HeLa cells and its possible regulation mechanism.Methods HeLa cells were treated with curcumin at 50μmol/L for 0,24,36,48 and 72 hours,respectively.MTT assay was used to observe the effects of curcumin on HeLa cell viability.Transwell assay was applied to detect the inhibitory effect of curcumin on the invasive metastatic ability of HeLa cells.Meanwhile,Western-blot was applied to detect the expression levels of metallo-matrix proteinase-2,9(MMP-2 and MMP-9)and brain-derived neurotrophic factor(BDNF)expression levels.The effects of curcumin on the expression of miR-206 and BDNF were examined by RT-qPCR and the regulatory relationship of miR-206 on BDNF by dual-luciferase assay.Results Curcumin significantly decreased the cell viability of HeLa cells in a time-dependent manner.Curcumin significantly reduced the invasion and migration ability of HeLa cells and was able to reduce the expression levels of MMP-2 and MMP-9,while activating the expression level of miR-206.In addition,dual luciferase results demonstrated that miR-206 inhibited the invasion and migration ability of HeLa cells by targeting the degradation of BDNF.Conclusion The miR-206/BDNF pathway activated by curcumin,which prevents certical cancer HeLa cells from migrating and invading.