College of Science, Sichuan Agricultural University, Ya'an, China.
College of Life Science, China West Normal University, Nanchong, China.
Cell Prolif. 2020 Feb;53(2):e12739. doi: 10.1111/cpr.12739. Epub 2019 Dec 9.
Tanshinone I (Tan-I) is one of the vital fatsoluble monomer components, which extracted from Chinese medicinal herb Salvia miltiorrhiza Bunge. It has been shown that Tan-I exhibited anti-tumour activities on different types of cancers. However, the underlying mechanisms by which Tan-Ⅰ regulates apoptosis and autophagy in ovarian cancer remain unclear. Thus, this study aimed to access the therapy effect of Tan-Ⅰ and the underlying mechanisms.
Ovarian cancer cells A2780 and ID-8 were treated with different concentrations of Tan-Ⅰ (0, 1.2, 2.4, 4.8 and 9.6 μg/mL) for 24 hours. The cell proliferation was analysed by CCK8 assay, EdU staining and clone formation assay. Apoptosis was assessed by the TUNEL assay and flow cytometry. The protein levels of apoptosis protein (Caspase-3), autophagy protein (Beclin1, ATG7, p62 and LC3II/LC3I) and PI3K/AKT/mTOR pathway were determined by Western blot. Autophagic vacuoles in cells were observed with LC3 dyeing using confocal fluorescent microscopy. Anti-tumour activity of Tan-Ⅰ was accessed by subcutaneous xeno-transplanted tumour model of human ovarian cancer in nude mice. The Ki67, Caspase-3 level and apoptosis level were analysed by immunohistochemistry and TUNEL staining.
Tan-Ⅰ inhibited the proliferation of ovarian cancer cells A2780 and ID-8 in a dose-dependent manner, based on CCK8 assay, EdU staining and clone formation assay. In additional, Tan-Ⅰ induced cancer cell apoptosis and autophagy in a dose-dependent manner in ovarian cancer cells by TUNEL assay, flow cytometry and Western blot. Tan-Ⅰ significantly inhibited tumour growth by inducing cell apoptosis and autophagy. Mechanistically, Tan-Ⅰ activated apoptosis-associated protein Caspase-3 cleavage to promote cell apoptosis and inhibited PI3K/AKT/mTOR pathway to induce autophagy.
This is the first evidence that Tan-Ⅰ induced apoptosis and promoted autophagy via the inactivation of PI3K/AKT/mTOR pathway on ovarian cancer and further inhibited tumour growth, which might be considered as effective strategy.
丹参酮 I（Tan-I）是从中药丹参中提取的一种重要脂溶性单体成分，已证明其对多种类型的癌症具有抗肿瘤活性。然而，Tan-Ⅰ调节卵巢癌细胞凋亡和自噬的潜在机制尚不清楚。因此，本研究旨在探讨 Tan-Ⅰ的治疗作用及其潜在机制。
用不同浓度的 Tan-Ⅰ（0、1.2、2.4、4.8 和 9.6μg/ml）处理卵巢癌细胞 A2780 和 ID-8 24 小时。用 CCK8 法、EdU 染色和克隆形成实验分析细胞增殖。用 TUNEL 法和流式细胞术检测细胞凋亡。用 Western blot 检测凋亡蛋白（Caspase-3）、自噬蛋白（Beclin1、ATG7、p62 和 LC3II/LC3I）和 PI3K/AKT/mTOR 通路蛋白的表达水平。用 LC3 染色观察细胞中的自噬小体。用裸鼠皮下异种移植卵巢癌模型评估 Tan-Ⅰ的抗肿瘤活性。用免疫组化和 TUNEL 染色分析 Ki67、Caspase-3 水平和凋亡水平。
CCK8 法、EdU 染色和克隆形成实验结果显示，Tan-Ⅰ呈剂量依赖性抑制卵巢癌细胞 A2780 和 ID-8 的增殖。此外，TUNEL 法、流式细胞术和 Western blot 结果显示，Tan-Ⅰ呈剂量依赖性诱导卵巢癌细胞凋亡和自噬。Tan-Ⅰ通过诱导细胞凋亡和自噬显著抑制肿瘤生长。机制上，Tan-Ⅰ通过激活凋亡相关蛋白 Caspase-3 的裂解促进细胞凋亡，抑制 PI3K/AKT/mTOR 通路诱导自噬。
这是第一个证明 Tan-Ⅰ通过抑制 PI3K/AKT/mTOR 通路激活诱导卵巢癌细胞凋亡和促进自噬，进而抑制肿瘤生长的证据，可能是一种有效的治疗策略。
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