隐丹参酮对人乳腺癌MCF7细胞的抑制作用及其机制

杨舒涵; 王玉琴; 刘航; 夏丽洁; 刘苏颖; 张英

doi:10.3971/j.issn.1000-8578.2023.23.0116

隐丹参酮对人乳腺癌MCF7细胞的抑制作用及其机制

Mechanism of Cryptotanshinone Inhibiting Proliferation of Human Breast Cancer MCF7 Cells

摘要

摘要:
目的探讨隐丹参酮（CPT）对人乳腺癌MCF7细胞的抑制作用及机制。
方法 MTT法检测CPT干预后MCF7细胞存活率，Annexin V/PI实验、Hoechst33258荧光染色实验检测细胞凋亡；流式细胞术检测细胞周期及细胞内活性氧ROS；细胞划痕实验、Transwell小室实验检测细胞迁移与侵袭；微球体培养及流式细胞术检测微球体表面分子CD44与CD24；Western blot检测细胞相关蛋白的表达。
结果 CPT呈剂量依赖性地抑制MCF7细胞增殖，24 h半数抑制浓度（IC₅₀）为19.24 μmol/L。与未加药处理组相比，CPT处理组可将细胞周期阻滞于S期，并诱导细胞凋亡。细胞划痕实验和Transwell小室结果表明，CPT对MCF7细胞的迁移与侵袭有显著抑制作用。此外，CPT降低了MCF7细胞来源微球体中CD24^−/lowCD44⁺细胞群。Western blot结果证明，CPT能明显上调Bax蛋白表达，下调Bcl2、PI3K-p85、Akt、N-cadherin、Twist1、Sox2、Oct4、Nanog蛋白表达，有效抑制ER-α磷酸化，降低了多重药物转运蛋白ABCG2的表达。
结论 CPT通过抑制MCF7细胞迁移与侵袭、降低CD24^−/low CD44⁺细胞群数量，影响肿瘤干细胞相关蛋白表达降低细胞干性，从而抑制细胞增殖。

Abstract:
Objective To investigate the inhibitory effect of cryptotanshinone (CPT) on human breast cancer cell MCF7 and its mechanism.
Methods The survival rate of MCF7 cells was measured by MTT assay. Cell apoptosis was detected by Annexin V/PI assay and Hoechst 33258 fluorescence staining assay. Cell cycle and reactive oxygen species were detected by flow cytometry. Cell migration and invasion were detected by cell scratch test and Transwell chamber test. The surface molecules CD44 and CD24 were detected by flow cytometry and microsphere culture. The expression of cell-associated proteins was detected by Western blot.
Results CPT inhibited the proliferation of MCF7 cells in a dose-dependent manner, and the 24 h IC₅₀ value was 19.24 μmol/L. Compared with the untreated group, the CPT-treated group showed cell cycle arrested in the S phase, and apoptosis was induced. The results of the cell scratch and Transwell chamber tests showed that CPT significantly inhibited the migration and invasion of MCF7 cells. Furthermore, CPT reduced the CD24^-/LowCD44⁺ cell population in MCF7 cell-derived microspheres. Western blot results showed that CPT could up-regulate the expression of Bax protein, down-regulate the expression of BCL-2, PI3K-p85, Akt, N-cadherin, Twist1, Sox2, Oct4, and Nanog protein, effectively inhibit the phosphorylation of ER-α, and decrease the expression of ABCG2.
Conclusion CPT can inhibit the proliferation of MCF7 cells by inhibiting the migration and invasion of MCF7 cells, decreasing the number of CD24^-/lowCD44⁺ cells and affecting the expression of tumor stem cell-related proteins.

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