Daidzein exerts anticancer activity towards SKOV3 human ovarian cancer cells by inducing apoptosis and cell cycle arrest, and inhibiting the Raf/MEK/ERK cascade
Authors: Chang-Hua Li
Fu Hua
Xiao-Gang Chen
Xiao-Ping Liu
Journal: International Journal of Molecular Medicine (2018)
Discipline: CLINICAL MEDICINE
Keywords: Ovarian cancer; Daidzein; Anticancer activity; Cell lines; Apoptosis
Affiliations: Department of Gynecology, Huai'an First People's Hospital, Nanjing Medical University, Huai'an, Jiangsu 223300, P.R. China., Department of Orthopaedics, Huai'an First People's Hospital, Nanjing Medical University, Huai'an, Jiangsu 223300, P.R. China.

Abstract:  Ovarian cancer is the main cause of gynecological cancer‑associated mortality around the world. Despite initial responses to chemotherapy, frequent relapse occurs. Daidzein is an important flavonoid and has been shown to exhibit a diversity of pharmacological properties, including antimicrobial and anticancer activities. However, information on the anticancer activity of daidzein against ovarian cancer remains limited. Therefore, the present study evaluated the anticancer activity of daidzein against a panel of human ovarian cancer cell lines and one normal ovarian cell line (Moody). The results revealed that daidzein exhibited potent anticancer activity against SKVO3 cells with a half‑maximal inhibitory concentration (IC50) of 20 µM. However, it exhibited comparatively lower activity against normal ovarian Moody cells, which had an IC50 of 100 µM. Daidzein induced morphological changes in SKOV3 cells and mitochondrial apoptosis, as evident from DAPI, AO/EB and Annexin V/propidium iodide staining. This was associated with the upregulation of B‑cell lymphoma 2‑associated X protein, cytochrome c, cleaved caspase‑3 and ‑9, and cleaved poly (ADP‑ribose) polymerase. Daidzein also triggered G2/M cell arrest through the downregulation of pCdc25c, Cdc25c, pCdc2, Cdc2 and cyclin B1. The effect of daidzein on the migration of SKOV3 cells was also determined, the results of which indicated that daidzein inhibited cell migration in a concentration‑dependent manner and was coupled with concomitant decrease in the expression of matrix metalloproteinase (MMP)‑2 and ‑9. Additionally, daidzein‑inhibited cell growth was simultaneous with suppression of the expression of phosphorylated mitogen‑activated protein kinase kinase and phosphorylated extracellular signal‑regulated kinase. The present study also examined whether daidzein exerts similar activity against SKOV3 cells in nude mouse xenograft models and it was revealed that daidzein considerably reduced the tumorigenesis in vivo, indicative of the potential for daidzein as a lead molecule in the development of ovarian cancer chemotherapy. 

Retracted: Yes

PubPeer comments:
 Our manuscript was published in 2018 and the study was performed long time ago. The article was submitted to different journals and ultimately ended up in International Journal of Molecular Medicine. I have moved to a different country in different institute since then Afterwards, if somebody copies our article wholly or in part, We surely are not responsible. , 
 *[right]* Figure 6 from [Hua et al (2018)] .  Effect of daidzein on the RAF MEK ERK signaling pathway. Effects of the indicated doses of daidzein on the expression of RAF MEK ERK signaling pathway proteins are shown . *[left]* Figure 7 from [Wang et al (2020)] .  Effect of indicated doses of sugiol on expression of p-MEK and p-ERK by western blotting. ![file] Different conditions, different loading controls, different authors. Same substantive bands. UPDATE: Another sighting! Figure 7 from [Ning et al (2019)] .  Effect of pinoresinol on the Ras MEK ERK signalling pathway as measured by western blotting analysis. The Figure depicts that pinoresinol blocks the Ras MEK ERK signalling pahway in ovarian cancer cells concentration-dependently. ![file] , 
 The earliest sighting so far is Fig 7B from [Li et al (2018)] .  Western blot analysis of the proteins expressed following treatment of cells with 3-tetrazolyl methyl-3-hydroxy-oxindole hybrid (THOH)... (B) MEK ERK signaling pathway proteins, detected by Western blot . ![file] , 
 #3 And that earlier appearance of the seemingly same set of bands were from Human Lung cell lines, per the other group of authors. And here per Hua, et al, its for ovarian cancer cells. ![file] , 
 *[right]* Figure 2C from [Hua et al (2018)] .  Effects of daidzein on apoptosis. Effects of the indicated doses of daidzein on ... (C) Determination of apoptotic cell populations using Annexin V PI staining in SKOV3 cells . *[left]* Figure 5 from [Ding Chen (2018)] .  Carvone induces apoptosis in KMS-5 myeloma cells as indicated by annexin V PI staining. The experiments were carried out in triplicate. The results indicate that exposure to 20u2009u03bcM of carbone leads to significant increase of apoptotic cells . ![file] , 
 *[right]* Figure 2B from [Hua et al (2018)] .  Effects of the indicated doses of daidzein on ... (B) AO EB staining . *[left]* Figure 4 from [Ding Chen (2018)] .  Carvone at IC50 triggers apoptosis in KMS-5 myeloma cells as indicated by AO EB staining. The experiments were carried out in triplicate. The green stained cells are normal cells while the orange stained cells are apoptotic cells. . ![file] , 
 *[right]* Figure 2A from [Hua et al (2018)] .  Effects of the indicated doses of daidzein on induction of apoptosis using (A) DAPI staining  . *[left]* Figure 3 from [Ding Chen (2018)] .  Carvone at IC50 triggers apoptosis in KMS-5 myeloma cells as indicated by DAPI staining. The experiments were carried out in triplicate. The bright fluorescent cells on the right image are apoptotic cells after carbone exposure. ![file] , 
 Figure 3 from [Hua et al (2018)] .  Effects of daidzein on MMP. Effects of the indicated doses of daidzein on (A) MMP, and the expression of (B) cyto c, Bax, Bclu20112 and Bid, (C) caspaseu20113, caspaseu20119, caspaseu20118 and PARP in SKOV3 cells. ![file] Figure 3 from [Zhou Hu (2018)] .  Effect of different concentrations of Evodiamine on the protein expression of apoptosis-related proteins as determined by Western blotting. ![file] UPDATE: Another sighting! Figure 2C,D from [An Liu (2020)] .  Camelliol C induces apoptosis in cervical cancer cells. D.. Western blots of Cyto C, Bax, Bcl-2, Bid (C), and caspase-3, -9, and -8 and PARP (D) . ![file] ------------------------------------------------ Some other manifestations of the same bands: Figure 8.  Effect of phillygenin on the expression of NF-kB expression. ![file] NF-kB band is unexpectedly similar to the Bcl-2 bands in *[left]* Figure 5 from [Luo et al (2019)] .  Effect of ferruginol on the expression apoptosis associated proteins as depicted by the western blot analysis . *[right]* Figure 6 from [Zhang et al (2019)] .  Effect of Trifolirhizin on the expression of Bax and Bcl-2 proteins as depicted by western blot analysis. The image shows that Trifolirhizin increased Bax and decreased Bcl-2 expression. ![file] , 
 #6 Fig 2B again *[right]* ![file] Another sighting! Figure 2 from [Yu, Zhou Li (2018)] .  Detection of apoptosis in untreated AGS, cisplatin-treated AGS, and cisplatin-treated AGS cells transfected with miR-136 by AO EB and DAPI staining. ![file] , 
 Retracted [7 APril 2021] .  Following the publication of the above paper, a concerned reader drew to the Editor s attention that several figures contained data that bore striking similarities to data published in other papers; notably, the western blot data shown in Fig. 6 appeared to have been presented in other studies, notably in Fig. 7B of another paper published around the same time and written by different authors based at different research institutions [Li P, Zhang Z, Zhang F, Zhou H and Sun W: Effects of 3-tetrazolyl methyl-3-hydroxy-oxindole hybrid (THOH) on cell proliferation, apoptosis, and G2 M cell cycle arrest occurs by targeting platelet-derived growth factor D (PDGF-D) and the MEK ERK signaling pathway in human lung cell lines SK-LU-1, A549, and A-427. *Med Sci Monit* 24: 4547-4554, 2018]. Furthermore, cellular images featured in Fig. 2A and B of the above paper appeared in Fig. 2 of the following paper, albeit the data were presented in a different field of view: Yu L, Zhou G-Q and Li D-C: MiR-136 triggers apoptosis in human gastric cancer cells by targeting AEG-1 and BCL2. *Eur Rev Med Pharmacol Sci* 22: 7251-7256, 2018.  After having conducted an independent investigation in the Editorial Office, the Editor of *International Journal of Molecular Medicine* has determined that this article should be retracted from the Journal on account of a lack of confidence concerning the originality and the authenticity of the data. The authors were asked for an explanation to account for these concerns, but the Editorial Office never received any reply. The Editor regrets any inconvenience that has been caused to the readership of the Journal. ,