Department of Gynecology, Huaihe Hospital of Henan University, 8 Baobei Road, Kaifeng, 475000, China.
Department of Gynecology, Huaihe Hospital of Henan University, 8 Baobei Road, Kaifeng, 475000, China.
Arch Biochem Biophys. 2020 Sep 30;691:108478. doi: 10.1016/j.abb.2020.108478. Epub 2020 Jul 23.
Calycosin, a functional phytoestrogen isoflavone isolated from Radix astragali, has been shown to possess multiple pharmacological properties including anti-cancer activity. However, up to now, the anti-cancer effect and the related mechanism of calycosin on cervical cancer (CC) cells have not been explored. It has been demonstrated that tumor suppressor miR-375 was downregulated in CC and calycosin upregulated miR-375 expression in cerebral ischemia/reperfusion. Thus we supposed that calycosin exerted anti-cancer effect by upregulating miR-375 expression in CC cells. Effects of calycosin or combined with miR-375 on cell viability and lactate dehydrogenase (LDH) release were detected by 3-(4,5)-dimethylthiahiazo (-z-y1)-3,5-di-phenytetra zoliumromide (MTT) and LDH release assay. Apoptosis, caspase-3 activity, and cell invasion were determined by flow cytometry, caspase-3 activity assay, and Transwell assay, respectively. miR-375 expression was detected by quantitative real-time PCR (qRT-PCR). Our results showed that Calycosin dose-dependently inhibited cell viability and increased LDH release in CC cells, suggesting the cytotoxic effect of calycosin on CC cells. Calycosin enhanced the apoptotic rate and caspase-3 activity and decreased the number of invaded cells in CC cells. In addition, we found that miR-375 expression was decreased in CC cells but was upregulated in response to calycosin. Mechanistically, knockdown of miR-375 significantly reversed the anti-cancer effect of calycosin on CC cells. In conclusion, calycosin inhibited viability, induced apoptosis, and suppressed invasion of CC cells by upregulating tumor suppressor miR-375.
毛蕊异黄酮是从黄芪中分离得到的一种功能性植物雌激素异黄酮，具有多种药理作用，包括抗癌活性。然而，到目前为止，毛蕊异黄酮对宫颈癌（CC）细胞的抗癌作用及其相关机制尚未得到探索。已经证明肿瘤抑制 miR-375 在 CC 中下调，而毛蕊异黄酮在上皮性卵巢癌细胞中上调 miR-375 的表达。因此，我们推测毛蕊异黄酮通过上调 CC 细胞中 miR-375 的表达发挥抗癌作用。通过 3-(4,5)-二甲基噻唑 (-z-y1)-3,5-二苯基四唑溴盐（MTT）和 LDH 释放测定法检测毛蕊异黄酮或与 miR-375 联合对细胞活力和乳酸脱氢酶（LDH）释放的影响。通过流式细胞术、caspase-3 活性测定和 Transwell 测定分别测定细胞凋亡、caspase-3 活性和细胞侵袭。通过定量实时 PCR（qRT-PCR）检测 miR-375 的表达。我们的结果表明，毛蕊异黄酮呈剂量依赖性抑制 CC 细胞活力并增加 LDH 释放，提示毛蕊异黄酮对 CC 细胞具有细胞毒性作用。毛蕊异黄酮增强了 CC 细胞的凋亡率和 caspase-3 活性，并减少了侵袭细胞的数量。此外，我们发现 miR-375 在 CC 细胞中的表达降低，但对毛蕊异黄酮的反应上调。从机制上讲，miR-375 的敲低显著逆转了毛蕊异黄酮对 CC 细胞的抗癌作用。总之，毛蕊异黄酮通过上调肿瘤抑制 miR-375 抑制 CC 细胞活力、诱导细胞凋亡和抑制细胞侵袭。
