摘要
目的探讨TLR4-NF-κb信号通路在小檗碱发挥抑制HeLa细胞增殖中的作用。方法分别给予5、10和40μg/mL 3种不同剂量干预Hela细胞,并通过特异性siRNA干扰TLR4表达。通过RT-qPCR和Western法检测TLR4、p65和Bcl-2的mRNA和蛋白表达改变,通过MTT法、Transwell小室和TUNEL评价细胞生长、迁移和凋亡情况。结果软件设计+RT-qPCR确定转染抑制效率最高的siRNA-002进行后续实验。小檗碱呈浓度依赖性抑制Hela细胞增殖和迁移,促进细胞凋亡,抑制p65、Bcl-2和TLR4的mRNA和蛋白表达。加用siRNA-002后,小檗碱的上述作用进一步增强。结论小檗碱可调控TLR4-NF-κB信号通路下调Bcl-2表达,显著抑制宫颈癌Hela细胞的增殖和迁移,促进其凋亡,为小檗碱治疗宫颈癌提供新的理论依据。
Objective To evaluate the effects of berberine on the proliferation of Hela cells through TLR4-NF-κB pathway. Methods Three different dose of berberine(5,10 and 40 μg/mL) and special TLR4-siRNA were used to intervene Hela cells. mRNA and protein of TLR4,p65 and Bcl-2 were evaluated by RT-qPCR and Western blot,respectively. The changes of proliferation,m igration and apoptosis of Hela cells were estimated by MTT,Transwell chamber and TUNEL,respectively. Results SiRNA-002 was selected as the strongest silence siR NA by RT-qPCR. The berberine could inhibit the proliferation and migration,promote the apoptosis,down regulate the mRNA and protein of p65,Bcl-2 and TLR4 of Hela cells,which showed a dose dependent effect. The effects of berberine above mentioned were enhanced by siRNA-002. Conclusion Berberine may inhibit Hela cells′ prolifera tion and migration,promote apoptosis by the down regulation of Bcl-2 through TLR4-NF-κB pathway.
作者
洪丹丹
任青玲
徐传花
徐雯雯
王伟
荣慧
郭红玉
赵玉芹
马蔚蓉
曹丹丹
陈冰倩
HONG Dandan;REN Qingling;XU Chuanhua;XU Wenwen;WANG Wei;RONG Hui;GUO Hongyu;ZHAO Yuqin;MA Weirong;CAO Dandan;CHEN Bingqian(Department of Gynecology,First Affiliated Hospital of Nanjing University of Chinese Medicine,Nanjing 210000,China)
出处
《实用医学杂志》
CAS
北大核心
2019年第5期692-697,共6页
The Journal of Practical Medicine
基金
江苏省中医院科技项目(编号:Y2017CX14)
第二周期省妇幼健康重点人才项目(省卫生计生委)(编号:FRC201711)
