金雀异黄素通过调控 FAK-paxillin 和 MAPK 信号通路抑制宫颈癌 HeLa 细胞的迁移和侵袭。

Genistein inhibits migration and invasion of cervical cancer HeLa cells by regulating FAK-paxillin and MAPK signaling pathways.

机构信息

Guangzhou University of Chinese Medicine, Guangzhou, 510405, China; Lingnan Medical Research Center of Guangzhou University of Chinese Medicine, Guangzhou, 510405, China; Guizhou University of Traditional Chinese Medicine, Guiyang, 550025, China.

Guangzhou University of Chinese Medicine, Guangzhou, 510405, China; Lingnan Medical Research Center of Guangzhou University of Chinese Medicine, Guangzhou, 510405, China.

出版信息

Taiwan J Obstet Gynecol. 2020 May;59(3):403-408. doi: 10.1016/j.tjog.2020.03.012.

DOI:10.1016/j.tjog.2020.03.012

PMID:32416888

Abstract

OBJECTIVE

Genistein obviously inhibits the migration and invasion of various tumor cells. However, its effects on cervical cancer cells have seldom been referred. We aimed to evaluate the effects of genistein on the proliferation, migration and invasion of cervical cancer HeLa cells, the expressions and phosphorylations of proteins related with FAK-paxillin and MAPKs signaling pathways, as well as the expressions of related key genes.

MATERIALS AND METHODS

HeLa cells were stimulated with genistein for 24 h and 48 h respectively. After adherence for 2 h, 0 μM, 12.5 μM, 25 μM, 50 μM and 100 μM genistein solutions were added in DMEM. Cell proliferation was tested by the CCK-8 assay. After treatment with 100 μM genistein, the migration ability was detected by the scratch assay. Transwell assay was used to detect cell migration and invasion abilities. Western blot and qRT-PCR were used to detect the expressions of proteins and mRNAs related with FAK-paxillin and MAPKs signaling pathways respectively.

RESULTS

The effect of genistein on the proliferation of HeLa cells was proportional to treatment time and drug dose, and the proliferation was inhibited after 24 h and 48 h at 100 μM. After treatment with 100 μM genistein, the scratch migration rate was significantly lower than that of the control group at 24 h and 48 h (P < 0.05). Genistein also inhibited the invasion of tumor cells through the upper chamber and Matrigel. The number of invasive cells was significantly lower than that of the control group (P < 0.05). Genistein significantly inhibited the phosphorylations of FAK, paxillin, p38 and p42/44. Compared to the control group, 100 μM genistein significantly suppressed the mRNA expressions of FAK, paxillin, Snail and twist.

CONCLUSION

Genistein inhibited the migration and invasion of cervical cancer HeLa cells by regulating FAK-paxillin and MAPK signaling pathways in dose-dependent manners.

摘要

目的

染料木黄酮明显抑制多种肿瘤细胞的迁移和侵袭。然而，其对宫颈癌的影响鲜有报道。本研究旨在评估染料木黄酮对宫颈癌 HeLa 细胞增殖、迁移和侵袭的影响，以及与 FAK-paxillin 和 MAPKs 信号通路相关的蛋白表达和磷酸化水平，以及相关关键基因的表达。

材料和方法

分别用染料木黄酮刺激 HeLa 细胞 24 h 和 48 h。贴壁 2 h 后，加入含 0 μM、12.5 μM、25 μM、50 μM 和 100 μM 染料木黄酮的 DMEM。用 CCK-8 法检测细胞增殖。用划痕实验检测 100 μM 染料木黄酮处理后细胞的迁移能力。Transwell 实验检测细胞迁移和侵袭能力。Western blot 和 qRT-PCR 分别检测与 FAK-paxillin 和 MAPKs 信号通路相关的蛋白和 mRNA 的表达。

结果

染料木黄酮对 HeLa 细胞增殖的抑制作用与作用时间和药物剂量呈正比，100 μM 作用 24 h 和 48 h 后细胞增殖受到抑制。用 100 μM 染料木黄酮处理后，24 h 和 48 h 的划痕迁移率明显低于对照组（P < 0.05）。染料木黄酮还通过上室和 Matrigel 抑制肿瘤细胞的侵袭。侵袭细胞数明显低于对照组（P < 0.05）。染料木黄酮显著抑制 FAK、paxillin、p38 和 p42/44 的磷酸化。与对照组相比，100 μM 染料木黄酮显著抑制 FAK、paxillin、Snail 和 twist 的 mRNA 表达。