Amentoflavone triggers cell cycle G2/M arrest by interfering with microtubule dynamics and inducing DNA damage in SKOV3 cells

Abstract

Ovarian cancer is the seventh most common cancer and the second most common cause of cancer-associated mortality among gynecological malignancies worldwide. The combination of antimitotic agents, such as taxanes, and the DNA-damaging agents, such as platinum compounds, is the standard treatment for ovarian cancer. However, due to chemoresistance, development of novel therapeutic strategies for the treatment of ovarian cancer remains critical. Amentoflavone (AMF) is a biflavonoid derived from the extracts of Selaginella tamariscina, which has been used as a Chinese herb for thousands of years. A previous study demonstrated that AMF inhibits angiogenesis of endothelial cells and induces apoptosis in hypertrophic scar fibroblasts. In order to check the influence of AMF on cell proliferation, the effects of AMF on cell cycle and DNA damage were measured by cell viability, flow cytometry, immunofluorescence and western blotting assays in SKOV3 cells, an ovarian cell line. In the present study, treatment with AMF inhibited ovarian cell proliferation, increased P21 expression, decreased CDK1/2 expression, interrupted the balance of microtubule dynamics and arrested cells at the G2 phase. Furthermore, treatment with AMF increased the expression levels of phospho-Histone H2AX (γ-H2AX; a variant of histone 2A, that belongs to the histone 2A family member X) and the DNA repair protein RAD51 homolog 1 (Rad51), indicating the occurrence of DNA damage since γ-H2AX and Rad51 are both key markers of DNA damage. Consistent with previous findings, the results of the present study suggest that AMF is a potential therapeutic agent for the treatment of ovarian cancer. In addition, the effects of AMF on cell cycle arrest and DNA damage induction may be the molecular mechanisms by which AMF might exert its potential therapeutic benefits in ovarian cancer.

Insights Into Amentoflavone: A Natural Multifunctional Biflavonoid

“…Increasing evidences demonstrate that AMF controls cell proliferation, apoptosis, invasion, metastasis, autophagy, transcription and drug-resistance in various types of cancers, such as lung cancer ( Banerjee et al, 2002a ; Banerjee et al, 2002b ; Jung et al, 2017 ; Hu et al, 2018 ; Park and Kim, 2019 ; Shen et al, 2019 ; Kim et al, 2020 ; Chen et al, 2021 ), cervical cancer ( Lee et al, 2011 ), ovarian cancer ( Liu et al, 2017a ; Zhang et al, 2020 ), bladder cancer ( Chiang et al, 2019 ), osteosarcoma ( Pan et al, 2017 ; Lee et al, 2019 ), melanoma ( Guruvayoorappan and Kuttan, 2007 ; 2008b ; a ; Siveen and Kuttan, 2011 ), breast cancer ( Lee et al, 2009 ; Pei et al, 2012 ; Lee et al, 2013 ; Chen et al, 2015 ; Aliyev et al, 2021 ), liver cancer ( Zheng et al, 2016 ; Chen et al, 2017a ; Lee et al, 2018a ; Lee et al, 2018b ; Tsai et al, 2018 ), brain cancer ( Yen et al, 2018 ; Zhaohui et al, 2018 ; Hsu et al, 2019 ; Chen et al, 2020c ), and oral squamous cell carcinoma ( Chen et al, 2020b ) via regulating kinds of signaling pathways ( Figure 2 ). These studies provide a lot of evidences that AMF is a potential effective multi-targeting drug for the prevention and treatment of a variety of cancers.…”

“…Additionally, the treatment of SiHa and CaSki cells with AMF induces cell cycle arrest at the sub-G1 phase through the down-regulation of p-pRb and G1/S cyclins and the up-regulation of p21 and p27 via a p53-dependent pathway ( Lee et al, 2011 ). Besides the effect of AMF on G1-phase cell cycle arrest, AMF treatment can inhibit cell proliferation, interrupt the balance of microtubule dynamics and arrest cells at the G2 phase via increasing p21 expression and decreasing CDK1/2 expression in ovarian cancer SKOV3 cells ( Zhang et al, 2020 ).…”

“…Followed that, AMF suppresses DNMT1 expression via the activation of ROS/AMPK and Sp1 signaling pathways ( Zhaohui et al, 2018 ). Moreover, in ovarian cancer cells AMF enhances the occurrence of DNA damage by increasing the expression levels of γ-H2AX and Rad51 ( Zhang et al, 2020 ).…”

Amentoflavone promotes ferroptosis by regulating reactive oxygen species (ROS) /5’AMP-activated protein kinase (AMPK)/mammalian target of rapamycin (mTOR) to inhibit the malignant progression of endometrial carcinoma cells

“…AF-induced cell cycle arrest and apoptosis could inhibit cell proliferation, which depended on mitochondrial pathway of breast cancer cells in vitro [ 8 ]. AF inhibited ovarian cell proliferation, interrupted microtubule dynamic balance, and stagnated cells in G2 phase [ 9 ]. However, the role of AF in the malignant progression of endometrial cancer has not been reported.…”

Design and Synthesis of Newly Synthesized Acrylamide Derivatives as Potential Chemotherapeutic Agents against MCF-7 Breast Cancer Cell Line Lodged on PEGylated Bilosomal Nano-Vesicles for Improving Cytotoxic Activity

“…Microtubules are also crucial for a variety of fundamental cell processes, including cell proliferation, sustained cell shape and structure, intracellular transport of vesicles and protein complexes and motility regulation [5][6][7]. Additionally, the disruption of microtubules can induce cell cycle arrest in the G2/M phase, formation of abnormal mitotic spindles and final triggering of signals for apoptosis [8][9][10]. Therefore, the importance of microtubules in mitosis and cell division makes them an attractive target for the development of anticancer drugs.…”