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, migration and apoptosis of Hela cells were estimated by MTT, Transwell chamber and TUN EL, respectively. Results SiRNA-002 was selected as the strongest silence siRNA 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' proliferation and migration, promote apoptosis by the down-regulation of Bcl-2 through TLR4-NF-κB pathway.
引文
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