m6A methyltransferase METTL3 promotes the progression of prostate cancer via m6A-modified LEF1
Authors: S-L Zhao
X-X Ma
Z-G Cao
Journal: European review for medical and pharmacological sciences (2020)
Discipline: PHARMACOLOGY & TOXICOLOGY
Keywords: METTL3; prostate cancer; N6-Methyladenosine; LEF1; mRNA
Affiliations: Department of Urology, Nanjing First Hospital, Nanjing Medical University, Nanjing, China. .

Abstract:  Objective: This study aims to uncover the regulatory effects of METTL3 on promoting the progression of prostate cancer (PCa) through N6-Methyladenosine (m6A) methylation on LEF1 mRNA. Patients and methods: The relative levels of METTL3 and LEF1 in 48 paired PCa tissues and adjacent ones were determined by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). Their correlation in PCa tissues was analyzed by Spearman correlation test. The survival of PCa patients affected by METTL3 was assessed by introducing Kaplan-Meier method. Wound closure assay was performed to evaluate the potential influence of METTL3 on migratory ability in PC-3 cells. Protein level of LEF1 in PC-3 cells with METTL3 or IGF2BP2 knockdown was examined. The activity of the Wnt pathway was tested through TOP/FOP-Flash. Furthermore, the interaction between LEF1 with METTL3 or IGF2BP2 was verified through RIP (RNA-Binding Protein Immunoprecipitation) assay. At last, the regulatory effects of METTL3/LEF1 axis on the activity of the Wnt pathway and migratory ability in PC-3 cells were determined. Results: METTL3 and LEF1 were upregulated in PCa tissues, and they presented a positive correlation in PCa. A high level of METTL3 predicted poor prognosis in PCa patients. The knockdown of METTL3 suppressed the migratory ability in PC-3 cells. Meanwhile, the knockdown of METTL3 downregulated protein level of LEF1 and decreased the activity of the Wnt pathway. The results of RIP assay indicated that METTL3 methylation sites were present on LEF1 mRNA. Moreover, the silence of METTL3 decreased the enrichment abundance of LEF1 in anti-IGF2BP2. Rescue experiments demonstrated that the overexpression of LEF1 partially reversed the regulatory effects of METTL3 on the Wnt activity and migratory ability in PCa cells. Conclusions: METTL3 is upregulated in PCa tissues. METTL3 influences the activity of the Wnt pathway through m6A methylation on LEF1 mRNA, thereafter, promoting the progression of PCa. 

Retracted: No

PubPeer comments:
 This paper is one of a set of (as of now) five papers that all share unexpected similarities. The papers are from different groups of authors at different hospitals, describing different cancer types. Yet, they share several similar features. * All five papers describe a group of 48 patients * All five papers describe the same set of GAPDH primers. One of them (the RV primer) does not appear to match the GAPDH mRNA sequence. * Figures 1 of all five papers show black bar graphs with error bars and a Kaplan-Meier curve with a blue and a red line * Two papers (XX Ma et al. and JL Xu et al.) share nearly identical positive correlation curves * All five papers show photos of wound healing (scratch) assays with the same background, suggesting they were all taken with the same microscope. Here is the set of papers: 1. J.-L. Xu, T. Hua, J. Ding, Y. Fan, Z.-J. Liu, J.-W. Lian - FOXF2 aggravates the progression of non-small cell lung cancer through targeting lncRNA H19 to downregulate PTEN - Eur Rev Med Pharmacol Sci 2019; 23 (24): 10796-10802 - DOI: 10.26355 eurrev_201912_19782 - Department of Oncology, Xi an Chest Hospital, Xi an, China 2. Q.-Z. Meng, C.-H. Cong, X.-J. Li, F. Zhu, X. Zhao, F.-W. Chen - METTL3 promotes the progression of nasopharyngeal carcinoma through mediating M6A modification of EZH2 - Eur Rev Med Pharmacol Sci 2020; 24 (8): 4328-4336 - DOI: 10.26355 eurrev_202004_21014 - Department of Head and Neck Surgery, Linyi Cancer Hospital, Linyi, China 3. T. Chen, J. Li, M.-H. Zhou, L.-J. Xu, T.-C. Pan - IL-6 stimulates lncRNA ZEB2-AS1 to aggravate the progression of non-small cell lung cancer through activating STAT1 - Eur Rev Med Pharmacol Sci 2020; 24 (7): 3734-3740 - DOI: 10.26355 eurrev_202004_20837 - Department of Cardiovascular Surgery, Tongji Hospital of Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China 4. T. Zhou, J.-J. Fang, Y.-X. Zhou, Z.-P. Li, L. Jiang, W.-W. Han, Z.-H. Zhu - Long non-coding RNA ANCR promotes progression of NSCLC by inhibiting E-Ca expression - Eur Rev Med Pharmacol Sci 2020; 24 (3): 1250-1257 - DOI: 10.26355 eurrev_202002_20178 - Department of Respiratory Medicine, Zhoushan Hospital of Zhejiang University, Zhoushan, China 5. X.-X. Ma, Z.-G. Cao, S.-L. Zhao - m6A methyltransferase METTL3 promotes the progression of prostate cancer via m6A-modified LEF1 - Eur Rev Med Pharmacol Sci 2020; 24 (7): 3565-3571 - DOI: 10.26355 eurrev_202004_20817 - Department of Urology, Nanjing First Hospital, Nanjing Medical University, Nanjing, China Here is a comparison of the Figures 1 from these papers. The very similar looking correlation curves of XX Ma et al. and JL Xu et al. have been highlighted with red boxes. ![file] Here is a comparison of the wound healing assay photos, with similar background spots highlighted in green and yellow boxes. Contrast enhanced. These appear to be irregularities in the microscope, which by itself is not a problem. But it is unexpected to see the same similarities in photos obtained across different laboratories. ![file] ![file] Could the authors please clarify how these photos look so similar? , 
  PCa tissues and adjacent normal ones (3 cm away from the tumor tissues) were surgically resected from 48 PCa patients undergoing surgical resection in Nanjing First Hospital from **April 2017 to December 2018** . ![file] If I understand correctly, at the time of the paper s publication in 2020, survival of patients had been tracked for 8 years after their recruitment, i.e. up to a period **April 2025 to December 2026**. , 
 Here is a superimposition of Figs 1D from this paper (in red) and 1D from [Xu et al (2019)] (in blue), to dramatise the degree of similarity. ![file] , 
 The 48 prostate cancer patients were recruited from April 2017-December 2018, presumably directly after their diagnosis, because they had not received anti-tumor therapy. Yet, despite the paper being published in 2020, the survival curve shows a 10-year (120 months) follow up. Something is not right here. HT: Hoya Camphorifolia For similar cases, see also: https: pubpeer.com publications B97218A01C913B80E7A89CDA87BDF7 and https: pubpeer.com publications 50C7E1823EED971D34431E8574D873 ![file] , 
 This paper is one of a set of (as of now) five papers that all share unexpected similarities. The papers are from different groups of authors at different hospitals, describing different cancer types. Yet, they share several similar features. * All five papers describe a group of 48 patients * All five papers describe the same set of GAPDH primers. One of them (the Reverse primer) does not appear to match the GAPDH mRNA sequence. * Figures 1 of all five papers show black bar graphs with error bars and a Kaplan-Meier curve with a blue and a red line * Two papers (XX Ma et al. and JL Xu et al.) share nearly identical positive correlation curves * All five papers show photos of wound healing (scratch) assays with the same background, suggesting they were all taken with the same microscope. Here is the set of papers: 1. J.-L. Xu, T. Hua, J. Ding, Y. Fan, Z.-J. Liu, J.-W. Lian - FOXF2 aggravates the progression of non-small cell lung cancer through targeting lncRNA H19 to downregulate PTEN - Eur Rev Med Pharmacol Sci 2019; 23 (24): 10796-10802 - DOI: 10.26355 eurrev_201912_19782 - Department of Oncology, Xi an Chest Hospital, Xi an, China 2. Q.-Z. Meng, C.-H. Cong, X.-J. Li, F. Zhu, X. Zhao, F.-W. Chen - METTL3 promotes the progression of nasopharyngeal carcinoma through mediating M6A modification of EZH2 - Eur Rev Med Pharmacol Sci 2020; 24 (8): 4328-4336 - DOI: 10.26355 eurrev_202004_21014 - Department of Head and Neck Surgery, Linyi Cancer Hospital, Linyi, China 3. T. Chen, J. Li, M.-H. Zhou, L.-J. Xu, T.-C. Pan - IL-6 stimulates lncRNA ZEB2-AS1 to aggravate the progression of non-small cell lung cancer through activating STAT1 - Eur Rev Med Pharmacol Sci 2020; 24 (7): 3734-3740 - DOI: 10.26355 eurrev_202004_20837 - Department of Cardiovascular Surgery, Tongji Hospital of Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China 4. T. Zhou, J.-J. Fang, Y.-X. Zhou, Z.-P. Li, L. Jiang, W.-W. Han, Z.-H. Zhu - Long non-coding RNA ANCR promotes progression of NSCLC by inhibiting E-Ca expression - Eur Rev Med Pharmacol Sci 2020; 24 (3): 1250-1257 - DOI: 10.26355 eurrev_202002_20178 - Department of Respiratory Medicine, Zhoushan Hospital of Zhejiang University, Zhoushan, China 5. X.-X. Ma, Z.-G. Cao, S.-L. Zhao - m6A methyltransferase METTL3 promotes the progression of prostate cancer via m6A-modified LEF1 - Eur Rev Med Pharmacol Sci 2020; 24 (7): 3565-3571 - DOI: 10.26355 eurrev_202004_20817 - Department of Urology, Nanjing First Hospital, Nanjing Medical University, Nanjing, China Here is a comparison of the Figures 1 from these papers. The very similar looking correlation curves of XX Ma et al. and JL Xu et al. have been highlighted with red boxes. ![file] Here is a comparison of the wound healing assay photos, with similar background spots highlighted in green and yellow boxes. Contrast enhanced. These appear to be irregularities in the microscope, which by itself is not a problem. But it is unexpected to see the same similarities in photos obtained across different laboratories. ![file] ![file] Could the authors please clarify how these photos look so similar? ,