Eriodictyol exerts potent anticancer activity against A549 human lung cancer cell line by inducing mitochondrial-mediated apoptosis, G2/M cell cycle arrest and inhibition of m-TOR/PI3K/Akt signalling pathway
Authors: Huanjuan Ni
Rui Zhang
Yong Zhang
Journal: Archives of Medical Science (2020)
Discipline: CLINICAL MEDICINE
Keywords: apoptosis; cell cycle arrest; eriodictyol; flow cytometer; lung cancer.
Affiliations: Department of Respiratory Medicine, The Affiliated Hospital of Panzhihua University, Panzhihua, China., Department of Respiratory Medicine, The First Affiliated Hospital of Chongqing Medical University, Chongqing, China., Department of Emergency, The Affiliated Hospital of Panzhihua University, Panzhihua, China.

Abstract:  Introduction: Eriodictyol is an important flavonoid and is commonly present across the plant kingdom. Flavonoids have been reported to show incredible pharmacological potential. However, the anticancer activity of the important flavonoid eriodictyol has not been well reported. In the present study we determined its anticancer potential against the human lung cancer cell line A549. Material and methods: The initial cytotoxicity induced by eriodictyol was measured by MTT assay. Flow cytometry was used to study the effects of eriodictyol on apoptosis, cell cycle phase distribution and mitochondrial membrane potential loss. The comet assay was used to measure DNA damage induced by eriodictyol in cancer cells while the western blot assay indicated effects of the compound on Bax/Blc-2 and PI3K/AKT/m-TOR proteins. Results: The results showed that eriodictyol has an IC50 value of 50 μM against human lung cancer cells as compared to the IC50 of 95 µM against non-cancerous FR2 cells. The molecule exerted its anticancer activity through induction of apoptosis by regulating the Bcl-2/Bax signalling pathway. It caused cell cycle arrest of human lung cancer A549 cells at G2/M phase. Eriodictyol was also found to cause a reduction of the mitochondrial membrane potential in a dose-dependent manner. Additionally, eriodictyol effectively inhibited the mTOR/PI3K/Akt signalling pathway in a dose-dependent manner. Conclusions: Based on the above findings, we conclude that eriodictyol exerts its anticancer activity through induction of mitochondrial apoptosis and G2/M cell cycle arrest and inhibition of the TOR/PI3K/Akt cascade, indicating that it may have potential as a lead compound in the treatment of lung cancer, provided further in depth studies are done. 

Retracted: No

PubPeer comments:
 The authors have made mistakes in quoting the references of table 1. Can authors provide erratum to the journal editor for providing correct information to readers ? Correct table is provided below with refernce number same as that in the article miRNAtEffect on EMTtTarget genetReference miR-10btPromotiontPTEN, HOXD10tHan et al., 14 Bahena-Ocampo et al.,33 Liang et al.34 and Ma et al.35 miR-21tPromotiontPDCD4, Tropomyosin, PTENtZadeh et al.,29 Petrovic,36 Yan et al.,37 Zhu et al.,38 Han et al.,39,40 Wu et al.,41 De Mattos-Arruda et al.42 miR-155tPromotiontFOXO3a, RhoAtYu et al.43 and Kong et al.44,45 miR-9tPromotiontE-cadherintKhew-Goodall et al.,46 Ma et al.47 miR-29atPromotiontTristetraprolintGebeshuber et al.48 miR- 103 107tPromotiontDicertMartello et al.49 miR-5003-3ptPromotiontMDM2tKwak et al.20 miR-181b-3ptPromotiontYWHAGtYoo et al.50 miR-221 222tPromotiontTRPS1tStinson et al.68 miR-183 96 182 ClustertPromotiontBRMS1L, GHRtZhang et al.69 miR-373tPromotiontTRPS1tHuang et al.70 miR-100tPromotiontSMARCA5tChen et al.51 miR-200 familytInhibitiontZEB1, ZEB2tZhang et al.,55 Bracken et al.,56 Lu et al.,57 Kong et al.,58 Wang and Ruan59 and Eger et al.60 miR-34 familytInhibitiontSnail, TPD52, Notch4tSiemens et al.61 Li et al.62 and Yu et al.63 miR-497tInhibitiontSmad7tLiu et al.18 miR-125btInhibitiontMAP2K7tHong et al.15 miR-206tInhibitiontTGF-u03b2, NRP1, Smad2tYin et al.64 miR-30atInhibitiontSlugtChang et al.65 miR-138tInhibitiontVimentintZhang et al.66 miR-195tInhibitiontFASN, HMGCR, ACACA, CYP27B1tSingh et al.67 miR-143tInhibitiontERK5tZhai et al.71 miR-671-5ptInhibitiontFOXM1tTan et al.72 miR-153tInhibitiontMetadherintLi et al.73 miR-300tInhibitiontTwisttYu et al.74 , 
 Dear Epichloe Clarkiiuff0cThank you for pointing out the errors in our paper. The mistakes in quoting the references were caused by changing the position of the table during the typesetting process. We immediately contacted the editorial board to correct it, but we did not receive a reply. At that time, this journal is in changing editorial board and publisher. From August to November, 2018, we continously communicated with the editorial board through emails for many times, and finally sent the erratum to the editorial board on November 5, 2018, but no further action has been taken since then. The following is the email information we exchanged. u53d1u4ef6u4ebauff1a7334714@qq.com t u65f6 u95f4uff1a2018u5e7411u67085u65e5(u661fu671fu4e00) u4e0au534810:04t u6536u4ef6u4ebauff1a Neisha Jobanputra Neisha.Jobanputra@sagepub.co.uk u9644 u4ef6uff1a 2 u4e2a ( Erratum and Response to the readeru2019s queries-20181029.doc...) t Dear Neisha, We attached our supplementary table and response again. Thanks Min Zhao ------------------ u539fu59cbu90aeu4ef6 ------------------ u53d1u4ef6u4eba:  Neisha Jobanputra Neisha.Jobanputra@sagepub.co.uk ; u53d1u9001u65f6u95f4: 2018u5e7410u670831u65e5(u661fu671fu4e09) u4e0bu53485:50 u6536u4ef6u4eba:  7334714 7334714@qq.com ; u6284u9001:  al19880206@163.com al19880206@163.com ;  756130482@qq.com 756130482@qq.com ;  66861197@qq.com 66861197@qq.com ; u4e3bu9898: RE: u56deu590duff1aRE: Reminder: Your Tumor Biology article - action required Dear Dr Zhao, Thank you for your reply. The Editor has assessed your response and notes that you have supplemented your table but have not described the added miRs in the text. Please describe the miRs supplemented in the table in the text as all other miRs have been. You should also refer to RNA-based therapeutics, as the comment from the reader reasonably suggested. Please send me your amended text on or before 7th November 2018. Best wishes, Neisha Neisha Jobanputra Publishing Editor SAGE Publishing 1 Oliveru2019s Yard, 55 City Road London, EC1Y1SP UK T: +44(0)20 7324 8787 www.sagepublishing.com SAGE Publishing is a trading name of SAGE Publications Ltd, Registered in England No.1017514 Los Angeles | London | New Delhi Singapore | Washington DC | Melbourne From: 7334714 7334714@qq.com Sent: 29 October 2018 12:00 To: Neisha Jobanputra Neisha.Jobanputra@sagepub.co.uk Cc: al19880206@163.com; 756130482@qq.com; 66861197@qq.com Subject: u56deu590duff1aRE: Reminder: Your Tumor Biology article - action required Dear Neisha, The attached files are our supplementary table and response. Thanks Min Zhao ------------------ u539fu59cbu90aeu4ef6 ------------------ u53d1u4ef6u4eba:  Neisha Jobanputra Neisha.Jobanputra@sagepub.co.uk ; u53d1u9001u65f6u95f4: 2018u5e7410u670826u65e5(u661fu671fu4e94) u665au4e0a7:32 u6536u4ef6u4eba:  7334714 7334714@qq.com ; u6284u9001:  zhao.min.hi@163.com zhao.min.hi@163.com ;  al19880206@163.com al19880206@163.com ;  756130482@qq.com 756130482@qq.com ;  66861197@qq.com 66861197@qq.com ; u4e3bu9898: RE: Reminder: Your Tumor Biology article - action required Dear Dr Zhao and colleagues, Thank you for providing your suggested erratum. Our apologies that you did not hear back from us previously. The Editor-in-Chief has assessed this and has requested you to provide further detail in your erratum text. The Editor has stated that the below highlighted section of the comments raised previously and some new additional feedback requires writing of two new paragraphs and supplementing the table with those miRs that refer to the EMT, and also supplementing your references. Please may we ask you to prepare this new, corrected text and table on or before Friday 2nd November. Please do acknowledge receipt of this email. Best wishes, Neisha Furthermore, authors missed out in quoting several important microRNAs that has been proved to regulate EMT, invasion and metastasis in breast cancer. Briefly, miR-7 [Target: SETDB1, STAT3; Inhibits migration invasion], miR-17 [Target: ROCK (Rho-associated kinase); promotes migration invasion], miR-19a-3p [Target: Fra1 (Fos-related antigen-1), STAT3; inhibits migration invasion], miR-24 [Target: NET1A; Inhibits migration invasion], miR-27 [Target: Nischarin, ST14 (Suppression of tumorigenicity 14); promotes migration invasion], miR-124 [Target: Slug, Flotillin-1; Inhibits migration invasion], miR-145 [Target: Mucin-1, JAM-A (Junctional adhesion molecule A, Mucin 1); Inhibits migration invasion], miR-146[Target: BRMS1 (Breast cancer metastasis suppressor 1); Inhibits migration invasion], miR-193b [Target: uPA (urokinase-type Plasminogen Activator); Inhibits migration invasion], miR-194 [Target: Talin2; Inhibits migration invasion], miR-205 [Target: ErbB3 and VEGF-A (vascular endothelial growth factor A); Inhibits migration invasion], miR-301a [Target: PTEN, FOXF2, Col2A1; Promotes migration invasion], miR-335[Target: Sox4 and tenascin C; Inhibits migration invasion], miR-489 [Target: Smad-3, ULK1, LAPTM4B; Inhibits migration invasion], miR-661[Target: MTA1 (metastatic tumor antigen 1); Inhibits migration invasion].2,3 Furthermore, adding to this ever increasing list includes a recent study which showed miR-22 to inhibit differentiation and metastasis of breast cancer cells by targeting ACLY (ATP citrate lyase).4 In the past two decades, RNA based therapeutics (miRNA and siRNA) have even entered into the clinical trials. So far till 2016, several miR mimics are in Phase III of clinical trials whereas none of the cases are reported for siRNAs (small interfering RNAs). Authors in their review did injustice to this important aspect by highlighting only about miR-34. However, Smith et al., in their very recent review article have extensively discussed about the observational clinical trials related to microRNAs and metastatic breast cancer. 5 The authors should provide the corrected and updated account of data related to microRNA target genes in breast cancer addressing the discrepancy present in the article by Zhao et al., along with the failure to incorporate few important miRNA. Neisha Jobanputra Publishing Editor SAGE Publishing 1 Oliveru2019s Yard, 55 City Road London, EC1Y1SP UK T: +44(0)20 7324 8787 www.sagepublishing.com SAGE Publishing is a trading name of SAGE Publications Ltd, Registered in England No.1017514 Los Angeles | London | New Delhi Singapore | Washington DC | Melbourne From: Neisha Jobanputra Sent: 04 October 2018 15:08 To: 7334714 7334714@qq.com Subject: RE: Reminder: Your Tumor Biology article - action required Dear Dr Zhao, Thank you for your reply. We will take your response into consideration and we will be in touch in due course. Best wishes, Neisha From: 7334714 7334714@qq.com Sent: 04 October 2018 04:35 To: Neisha Jobanputra Neisha.Jobanputra@sagepub.co.uk Subject: Re: Reminder: Your Tumor Biology article - action required For the reference errors quoted in Table 1, we gave the Erratum letter on Date: 2017-07-28 14:16:59., as show in the following. Now we send the Erratum again. -------- Forwarding messages -------- From:  u8d75u654f zhao.min.hi@163.com Date: 2017-07-28 14:16:59 To: francesca.marr@sagepub.com Subject: Erratum for reference errors in Table 1 Dear Editor-in-Chief and Dr. Vikas Yadav, We are very sorry for the reference errors quoted in Table 1. The attached file is our corrected Table 1. We throughly checked the order of the cited references, and we hope the corrected table will be right for Erratum. Thank you very much. Best wishes Min Zhao ------------------ Original ------------------ From:  Neisha Jobanputra Neisha.Jobanputra@sagepub.co.uk ; Date: Tue, Oct 2, 2018 06:56 PM To:  zhao.min.hi@163.com zhao.min.hi@163.com ; Cc:  al19880206@163.com al19880206@163.com ;  756130482@qq.com 756130482@qq.com ;  66861197@qq.com 66861197@qq.com ;  7334714 7334714@qq.com ; Subject: Reminder: Your Tumor Biology article - action required Dear Dr Zhao, This is a brief reminder that we are currently awaiting your response on the below queries raised on your article. Please respond to us as soon as possible. Please note that if we do not receive a response from you shortly then this will be escalated to your institution. Best wishes, Neisha Neisha Jobanputra Publishing Editor SAGE Publishing 1 Oliveru2019s Yard, 55 City Road London, EC1Y1SP UK T: +44(0)20 7324 8787 www.sagepublishing.com SAGE Publishing is a trading name of SAGE Publications Ltd, Registered in England No.1017514 Los Angeles | London | New Delhi Singapore | Washington DC | Melbourne From: Neisha Jobanputra Sent: 20 September 2018 10:48 To: zhao.min.hi@163.com Subject: Your Tumor Biology article - action required Dear Dr Zhao, I hope this finds you well. I am writing from SAGE Publishing regarding your Tumor Biology article u201cMicroRNAs regulate the epithelial-mesenchymal transition and influence breast cancer invasion and metastasisu201d. We have received the below correspondence from a reader regarding your article, and so please may we ask you to provide a response to these points raised, by email, on or before Friday 28th September? Please do not hesitate to contact me if you have any queries, and I look forward to hearing from you shortly. Best wishes, Neisha ******* The review article appears to be well written providing us a summary of work done in the field of microRNA based therapeutics for breast cancer treatment. However in order to avoid misinterpretation of valued information quoted in the article and to acknowledge the achievement by other groups working for common cause, I intend to correct and update the information, based on the previous original version published in your journal. The authors have inadvertently made mistakes in quoting the references of table 11, which has now been rectified in this letter.Furthermore, authors missed out in quoting several important microRNAs that has been proved to regulate EMT, invasion and metastasis in breast cancer. Briefly, miR-7 [Target: SETDB1, STAT3; Inhibits migration invasion], miR-17 [Target: ROCK (Rho-associated kinase); promotes migration invasion], miR-19a-3p [Target: Fra1 (Fos-related antigen-1), STAT3; inhibits migration invasion], miR-24 [Target: NET1A; Inhibits migration invasion], miR-27 [Target: Nischarin, ST14 (Suppression of tumorigenicity 14); promotes migration invasion],miR-124 [Target: Slug, Flotillin-1; Inhibits migration invasion], miR-145 [Target: Mucin-1, JAM-A (Junctional adhesion molecule A, Mucin 1); Inhibits migration invasion], miR-146 [Target: BRMS1 (Breast cancer metastasis suppressor 1); Inhibits migration invasion], miR-193b [Target: uPA (urokinase-type Plasminogen Activator); Inhibits migration invasion], miR-194 [Target: Talin2; Inhibits migration invasion], miR-205 [Target: ErbB3 and VEGF-A (vascular endothelial growth factor A); Inhibits migration invasion], miR-301a [Target: PTEN, FOXF2, Col2A1; Promotes migration invasion], miR-335 [Target: Sox4 and tenascin C; Inhibits migration invasion], miR-489[Target: Smad-3, ULK1, LAPTM4B; Inhibits migration invasion], miR-661 [Target: MTA1 (metastatic tumor antigen 1); Inhibits migration invasion].2,3 Furthermore, adding to this ever increasing list includes a recent study which showed miR-22 to inhibit differentiation and metastasis of breast cancer cells by targeting ACLY (ATP citrate lyase).4 Table 1. Key miRNAs associated with EMT regulation in breast cancer; Note- reference numbers quoted in table corresponds to the references in article (Zhao et al., 2017). miRNAtEffect on EMTtTarget genetReference miR-10btPromotiontPTEN, HOXD10tHan et al., 14 Bahena-Ocampo et al.,33 Liang et al.34 and Ma et al.35 miR-21tPromotiontPDCD4, Tropomyosin,PTENtZadeh et al.,29 Petrovic,36 Yan et al.,37 Zhu et al.,38 Han et al.,39,40Wu et al.,41 De Mattos-Arruda et al.42 miR-155tPromotiontFOXO3a, RhoAtYu et al.43 and Kong et al.44,45 miR-9tPromotiontE-cadherintKhew-Goodall et al.,46 Ma et al.47 miR-29atPromotiontTristetraprolintGebeshuber et al.48 miR- 103 107tPromotiontDicertMartello et al.49 miR-5003-3ptPromotiontMDM2tKwak et al.20 miR-181b-3ptPromotiontYWHAGtYoo et al.50 miR-221 222tPromotiontTRPS1tStinson et al.68 miR-183 96 182ClustertPromotiontBRMS1L, GHRtZhang et al.69 miR-373tPromotiontTRPS1tHuang et al.70 miR-100tPromotiontSMARCA5tChen et al.51 miR-200 familytInhibitiontZEB1, ZEB2tZhang et al.,55 Bracken et al.,56 Lu et al.,57 Kong et al.,58 Wang and Ruan59 and Eger et al.60 miR-34 familytInhibitiontSnail, TPD52, Notch4tSiemens et al.61 Li et al.62 and Yu et al.63 miR-497tInhibitiontSmad7tLiu et al.18 miR-125btInhibitiontMAP2K7tHong et al.15 miR-206tInhibitiontTGF-u03b2, NRP1, Smad2tYin et al.64 miR-30atInhibitiontSlugtChang et al.65 miR-138tInhibitiontVimentintZhang et al.66 miR-195tInhibitiontFASN, HMGCR, ACACA, CYP27B1tSingh et al.67 miR-143tInhibitiontERK5tZhai et al.71 miR-671-5ptInhibitiontFOXM1tTan et al.72 miR-153tInhibitiontMetadherintLi et al.73 miR-300tInhibitiontTwisttYu et al.74 In the past two decades, RNA based therapeutics (miRNA and siRNA) have even entered into the clinical trials. So far till 2016, several miR mimics are in PhaseIII of clinical trials whereas none of the cases are reported for siRNAs (small interfering RNAs). Authors in their review did injustice to this important aspect by highlighting only about miR-34. However, Smith et al., in their very recent review article have extensively discussed about the observational clinical trials related to microRNAs and metastatic breast cancer. 5 Reference 1. Zhao M, Ang L, Huang J, Wang J. MicroRNAs regulate the epithelial-mesenchymal transition and influence breast cancer invasion and metastasis. Tumor Biol 2017; 39, 1010428317691682 2. Kaboli PJ, Rahmat A, Ismail P, Ling KH. MicroRNA based therapy and breast cancer: A comprehensive review of novel therapeutic strategies from diagnosis to treatment. Pharmacol Res 2015; 97, 104-21 3. Le Quesne J, Caldas C. Micro-RNAs and breast cancer. Mol Oncol 2010; 4, 231-40. 4. Liu H, Huang X, Ye T. MiR-22 down-regulates the proto-oncogene ATP citrate lyase to inhibit the growth and metastasis of breast cancer. Am J Transl Res 2018; 10, 659-69. 5. Smith B, Agarwal P, Bhowmick NA. MicroRNA applications for prostate, ovarian and breast cancer in the era of precision medicine.Endocr Relat Cancer 2017; 24, R152-R172. ***** Neisha Jobanputra Publishing Editor SAGE Publishing 1 Oliveru2019s Yard, 55 City Road London, EC1Y1SP UK T: +44(0)20 7324 8787 www.sagepublishing.com SAGE Publishing is a trading name of SAGE Publications Ltd, Registered in England No.1017514 Los Angeles | London | New Delhi Singapore | Washington DC | Melbourne ,