慢性氧化应激激活GSK3β/β-catenin信号通路增强乳腺癌细胞MCF-7活性的机制研究
Mechanical Research of Chronic Oxidative Stress on the Activity of Breast Cancer Cells MCF-7 by Activating GSK3β/β-catenin Signaling Pathway
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摘要: 目的 建立MCF-7乳腺癌细胞慢性氧化应激细胞模型，观测MCF-7乳腺癌细胞在慢性氧化应激下的增殖情况，探讨慢性应激状态对乳腺癌增殖、迁移和侵袭的影响及其可能机制。方法 CCK-8法检测MCF-7分别在急性氧化应激和慢性氧化应激干预的情况下的增殖抑制率，通过划痕实验和transwell实验检测慢性氧化应激条件下MCF-7细胞迁移和侵袭的能力。通过ELISA法测定IL-6水平，检测慢性氧化应激模型条件下MCF-7的超氧化物歧化酶活性（SOD）和总抗氧化能力（T-AOC）。qPCR及Western blot检测GSK3β/β-catenin信号通路相关蛋白及RNA表达水平。结果 与急性氧化应激比较，慢性氧化应激条件下，MCF-7的增殖能力增强，其SOD和T-AOC的能力也进一步增强。其迁移和侵袭的能力均强于野生型和急性氧化应激下的MCF-7。该结果和IL-6蛋白表达，GSK3β/β-catenin信号通路相关蛋白和RNA表达相一致。同时发现三黄煎剂能够抑制MCF-7慢性氧化应激细胞的增殖。结论 慢性氧化应激状态能够促进MCF-7乳腺癌细胞增殖和转移，其机制可能与GSK3β/β-catenin信号通路激活和IL-6表达增加相关。
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关键词:
- 乳腺癌MCF-7细胞 /
- 慢性氧化应激 /
- GSK3β/β-catenin信号通路 /
- IL-6 /
- 三黄煎剂
Abstract: OBJECTIVE To establish a oxidative stress cell line model of breast cancer cells MCF-7， observe the growth under this environment， and explore the effects of chronic stress status on breast cancer proliferation， migration and invasion of breast cancer and its possible mechanisms. METHODS The CCK-8 assay was used to detect the proliferation inhibition rate of MCF-7 under the conditions of acute oxidative stress and chronic oxidative stress. The ability of MCF-7 cells to migrate and invade under chronic oxidative stress conditions was examined by scratch assay and transwell assay. The level of IL-6 was determined by ELISA. Superoxide dismutase activity (SOD) and total antioxidant capacity (T-AOC) of MCF-7 under chronic oxidative stress model conditions were tested by kit. The expression levels of GSK3β/β-catenin signaling pathway-related proteins and RNA were determined by RT-PCR and Western blot. RESULTS Compared with acute oxidative stress， under the condition of chronic oxidative stress， the value-adding ability of MCF-7 was enhanced， and the ability of SOD and T-AOC was further enhanced. Its ability to migrate and invade is stronger than that of wild-type and MCF-7 under acute oxidative stress. This result is consistent with IL-6 protein expression， GSK3β/β-catenin signaling pathway-associated protein and RNA expression. It was also found that Sanhuang Decoction can inhibit the proliferation of MCF-7 chronic oxidative stress cells. CONCLUSION Chronic oxidative stress can promote the proliferation and metastasis of breast cancer cells MCF-7， and its mechanism may be related to the activation of GSK3β/β-catenin signaling pathway and the increase of IL-6 expression. -
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