Mechanism of MicroRNA-708 Targeting BAMBI in Cell Proliferation, Migration, and Apoptosis in Mice With Melanoma via the Wnt and TGF-β Signaling Pathways
Authors: Dong-Mei Wu
Hong-Jie Lu
Hai-Lin Zhang
Jing Yan
Jun Lu
Gui-Hong Zheng
Ming Bai
Pei-Ying Jin
Yuan-Lin Zheng
Journal: Technology in Cancer Research & Treatment (2018)
Discipline: CLINICAL MEDICINE
Keywords: BAMBI gene; TGF-β signaling pathway; Wnt signaling pathway; apoptosis; melanoma
Affiliations: 1 Key Laboratory for Biotechnology on Medicinal Plants of Jiangsu Province, School of Life Science, Jiangsu Normal University, Xuzhou, People's Republic of China., 2 College of Health Sciences, Jiangsu Normal University, Xuzhou, People's Republic of China., 3 Emergency Center, The Affiliated Hospital of Xuzhou Medical University, Xuzhou, People's Republic of China., 4 Department of Plastic and Reconstructive Surgery, Peking Union Medical College Hospital, Chinese Academy of Medical Science and Peking Union Medical College, Beijing, People's Republic of China.

Abstract:  Objective: The aim of this study was to evaluate the mechanisms involved with miRNA-708 and its targeting of bone morphogenetic protein and activin membrane-bound inhibitor in cell proliferation, migration, and apoptosis in mice with melanoma via the Wnt and transforming growth factor β signaling pathways. Methods: Sixty mice were recruited of which 40 were subsequently assigned into the experimental group (22 mice were successfully established as melanoma model and 18 mice used in tumor xenograft), and the normal control group consisted of 20 mice. B16 cells were assigned to the normal, blank, and negative control, miR-708 mimics, miR-708 inhibitors, si-BAMBI, and miR-708 inhibitors + si-bone morphogenetic protein and activin membrane-bound inhibitor groups. Western blotting and reverse transcription quantitative polymerase chain reaction were employed to detect the expression levels within the tissues and cell lines. TCF luciferase reporter (TOP-FLASH) or a control vector (FOP-FLASH) was applied to detect the activity of the Wnt signaling pathway. MTT3-(4,5-Dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide assay, flow cytometry, scratch test, and Transwell assay were conducted, respectively, for cell proliferation, apoptosis, migration, and invasion, while tumor xenograft procedures were performed on the nude mice recruited for the study. Results: Compared to the normal control group, the model group displayed increased expressions of bone morphogenetic protein and activin membrane-bound inhibitor, Wnt10B, P53, and Bcl-2; TOPflash activity; β-catenin expression; cell proliferation; migration; and invasion capabilities while decreased expressions of miR-708, vascular endothelial growth factor, Fas, Bax, Caspase-3, and cleaved Caspase-3 and apoptosis rate. Compared to the blank and negative control groups, the miR-708 mimics and small-interfering RNA-bone morphogenetic protein and activin membrane-bound inhibitor groups exhibited decreases expressions of bone morphogenetic protein and activin membrane-bound inhibitor, Wnt10B, P53, and Bcl-2 and decreased proliferation, migration, and invasion capabilities, while increases in the apoptosis rate, expressions of vascular endothelial growth factor, Fas, Bax, Caspase-3, and cleaved Caspase-3; however, downregulated levels of TOPflash activity and β-catenin expression were recorded. The miR-708 inhibitors group displayed an opposite trend. Conclusion: Downregulation of miR-708-targeted bone morphogenetic protein and activin membrane-bound inhibitor inhibits the proliferation and migration of melanoma cells through the activation of the transforming growth factor β pathway and the suppression of Wnt pathway. 

Retracted: No

PubPeer comments:
 Figure 13. Transwell results (A, u00d7 200) ... in each group ![] siRNA-BAMBI and miR-708-mimic appear to be the same other than a 180-deg rotation. Blank, miR-708-inhibitor + siRNA-BAMBI, and NC (after a 180-deg rotation) substantially overlap, with the addition or substraction of some cells. , 
 *[left]* Figure 7C.  Expressions of miR-708, BAMBI, Wnt10B, P53, Bcl-2, VEGF, Fas, Bax, Caspase-3, and cleaved Caspase-3 after transfection in each group.... C, electrophoretograms of protein expressions . *[right]* Fig. 3C from [Jiang et al (2018)] .  The miR-139-5p, LPAR1, LPAR2, LPAR3, LPAR4, LPAR5, PI3K, Akt, E-cadherin, N-cadherin, Vimentin, TGF-u03b21 and CTGF mRNA and protein expressions in the IC group and the normal group ... C, image for Western blotting; D, LPAR1, LPAR2, LPAR3, LPAR4 and LPAR5 protein expressions . ![file] No authors in common. How to explain the resemblance within the loading controls? Now comparing 7C with 6C from Jiang et al. The resolution is not high but enough to see that the loading control bands are the same. ![file] , 
 Figure 14B.  Tumor ... size (B) of nude mice among each group. ![file] Two tumors seem to be identical apart from rotation and rescaling. Fig 7C from [Wen et al (2017)] .  Tumor volume and weight of nude mice of EC9706 cell line in the four groups after EC cell implantation ... (C) Transplanted tumors in the four groups. Horizontally-flipped version below. ![file] Relative sizes, cell-line, treatments and other details differ, but these are unmistakably the same tumours. , 
 Whoops, missed another duplicated and slightly-rotated tumor. Here, red boxes. ![file] Other color boxes indicate tumors also found in Fig 12B of [Zhang et al (2017)] , after horizontal flip or 180-deg rotation: ![file] , 
 #2 Fig 7C again, and 6C of Jiang et al (2018): ![] Another sighting of the loading control, this time labeled as beta-Actin rather than as GAPDH. Fig 8A from [Wu et al (2018)] . ![] , 
 #1 Fig 13A again: ![file] Conceivably linked in some way to Fig. 6 from [Zhang et al (2017)] ,  Effects of interfered or overexpressed long noncoding RNA-ROR on the invasion of breast cancer cells . ![file] UPDATE: Another sighting! Fig Fig. 4A,B from [Bai et al (2016)] .  Comparisons of migratory and invasive cell numbers among the blank, NC, p16INK4A and p14ARF groups. ... A Transwell migration of cells in each groups; B Transwell invasion of cells in each groups . ![] , 
 #4 Fig 14B again. ![] Another sighting of the tumors: Fig 11A from [He et al (2018)] .  Effect of different CUL4B expression levels on PC liver metastasis in nude mice Note: A, comparison of tumor formation and weight of the tumor in nude mice 4 weeks later . ![] And another! Fig 2B from [Wang et al (2017)] : ![file] And another! Fig 6D from [Wu et al (2018)] : ![file] Fig 6B from [Ji et al (2019)] . ![] , 
 Assessed here, as part of the first of 10 broad investigations: https: www.nsfc.gov.cn publish portal0 tab442 info85495.htm **Decisions on handling scientific research misconduct cases investigated and dealt with in 2022 (first batch)** Date 2022-05-13 Source: u3000 Author: u3000 u3010 Large, Middle and Small u3011 u3010Printu3011 u3010Closeu3011 u3000u3000The National Natural Science Foundation of China resolutely implements the spirit of the  Several Opinions on Further Strengthening the Construction of Scientific Research Integrity and  Opinions on Further Promoting the Spirit of Scientists and Strengthening the Construction of Style and Study Style by the General Office of the Central Committee of the Communist Party of China and the General Office of the State Council. Horizontal science and technology are self-reliant and self-reliant, continue to carry out scientific research integrity construction, and seriously investigate and deal with scientific research misconduct involving the National Natural Science Foundation of China. The Supervision Committee of the National Natural Science Foundation of China conducts investigations, verifications and comprehensive judgments on the accepted scientific research integrity cases, in accordance with the Regulations of the National Natural Science Foundation of ) and  Measures for Investigation and Handling of Scientific Research Misconduct in National Natural Science Foundation of China and other regulations and systems to propose handling suggestions. After deliberation at the second executive meeting of the National Natural Science Foundation of China in 2022, it was decided to deal with the relevant parties involved. The 10 handling decisions (online release version) that have been criticized and criticized are hereby announced. -------------------------------- About the existence of papers published on Wu Dongmei, Zheng Yuanlin, Lu Jun, etc. - Falsification of pictures or data, fabrication of research process, use of others signatures without consent, and false information in project final reports - processing decision - National Science and Technology Financial Supervisory Office [2022] No. 1 u3000u3000The Supervision Committee of the National Natural Science Foundation of China launched an investigation into several papers of Jiangsu Normal University, including Wu Dongmei, Zheng Yuanlin, and Jun Jun, for suspected academic misconduct. The papers involved are as follows: ... u3000u3000Paper 7:  Hong-Jie Lu # , Jing Yan # , Pei-Ying Jin # , Gui-Hong Zheng # , Hai-Lin Zhang # , Ming Bai # , Dong-Mei Wu*, Jun Lu*, and Yuan-Lin Zheng*. **Mechanism of MicroRNA-708 Targeting BAMBI in Cell Proliferation, Migration, and Apoptosis in Mice With Melanoma via the Wnt and TGF-u03b2 Signaling Pathways**. Technology in Cancer Research Treatment, 2018, 17:1-15.u201d (noted Fund numbers 81271225, 31201039, 81171012, 30950031, 81571055, 81400902) ... u3000u3000According to the investigation of the supporting unit, the above 35 papers involved all have the problem of improper use of pictures or data fraud, and the pictures or data in question are all provided by third-party companies. Among the 35 papers involved, 34 (except paper 21) still have the problem of using others signatures without consent, and papers 4, 5, 9, 12, 30, and 34 also have the problem of fabricating the research process. Wu Dongmei is the author of the above 35 papers involved, of which 12 papers (papers 2, 8, 9, 10, 11, 12, 14, 15, 25, 26, 30, 35) are the first author and 22 papers (Paper 1, 4, 5, 6, 7, 13, 16, 17, 18, 19, 20, 21, 22, 23, 24, 27, 28, 29, 31, 32, 33, 34) Corresponding author, They should be primarily responsible for the objective results of falsifying pictures or data, using others signatures without consent, and fabricating research processes; Zheng Yuanlin and Lu Jun, as the co-corresponding authors of the 35 papers involved, were negligent in reviewing and took secondary responsibility. In addition, Wu Dongmei included 4 papers involved (papers 21, 24, 28, 29) in the final report of the National Natural Science Foundation of China (Grant No. 81400902), and Zheng Yuanlin included 9 papers involved (papers 6, 8, 10, 13, 21, 24, 26, 28, 29) included in the final report of the National Natural Science Foundation of China (Grant No. 81571055), and should be responsible for the objective results of false information in the final report. u3000u3000After deliberation at the 12th meeting of the Fifth Supervision Committee of the National Natural Science Foundation of China (Life Medicine Professional Committee) and the approval of the second executive meeting of the National Natural Science Foundation of China in 2022, it was decided to follow the thirty-fifth National Natural Science Foundation of China Regulations. Article 4, Article 17, Items 3, 4, and 9 of  Measures for Handling Misconduct in Science Fund Work by the Supervision Committee of the National Natural Science Foundation of China (for Trial Implementation) , revocation of Wu Dongmei s National Natural Science Foundation of Molecular mechanism of TRPV4-mediated domoic acid treatment-induced cognitive deficits in mice (Grant No. 81400902), recovering the allocated funds, permanently disqualifying Wu Dongmei from the National Natural Science Foundation of China (since January 18, 2022), Give Wu Dongmei a circular of criticism; decide to follow Article 35, item 4 of the  National Natural Science Foundation of China Regulations , and refer to  The National Natural Science Foundation of China Supervision Committee s Handling of Misconduct in Science Fund Work (Trial) Tenth Items 3 and 4 of seven articles, the Zheng Yuanlin National Natural Science Foundation of China project  The Mechanism of HDAC6 Activating HMGB1 RAGE Signaling Axis to Mediate Cognitive Defects in Type 2 Diabetic Mice (Approval No. 81571055), recover the allocated funds, cancel Zheng Yuanlin will be eligible to apply for the National Natural Science Foundation of China for 5 years (January 18, 2022 to January 17, 2027), and will give Zheng Yuanlin a notice of criticism; it is decided to refer to the  National Natural Science Foundation of China Supervision Committee on misconduct in the work of the National Natural Science Foundation of China. Items 3 and 4 of Article 17 of the Measures (for Trial Implementation), the Army s National Natural Science Foundation of China project application qualification will be cancelled for 5 years (January 18, 2022 to January 17, 2027), and the Army will be criticized in a circular. - National Natural Science Foundation of China - March 30, 2022 , 
 Retracted [6 October 2022] . -------------------- At the request of the Executive Editor the following article has been retracted. Following article publication, two authors, Yuanlin Zheng and Guihong Zheng, contacted the journal editorial office stating that they were not aware of the publication and requested the article be retracted. While some authors have claimed no knowledge of this article, we are unable to find conclusive evidence to support this claim. Furthermore, multiple image duplications were identified both within figures of the article and figures of other articles, which is documented in comments on PubPeer. The authors were unable to provide the raw data and explained that they used a technical company to help generate data. The authors did not respond to further questions. Due to the lack of raw images this calls into question the validity of the findings and the Executive Editor of Technology in Cancer Research Treatment retracts the article. The institution contacted the journal to request a retraction, after the decision to retract had been made. ,