Decreased miR‐200a‐3p is a key regulator of renal carcinoma growth and migration by directly targeting CBL
Authors: Meng Ding
Cuiping Zhang
Yaping Tian
Jun-Jun Wang
Chunni Zhang
Jingping Ge
Ke Zen
Xinlei Sun
Chen-Yu Zhang
Jinsha Zhong
Cheng Wang
Journal: Journal of Cellular Biochemistry (2018)
Discipline: MOLECULAR BIOLOGY & GENETICS
Keywords: apoptosis; miR-200a-3p; migration; proliferation; renal cell carcinoma.
Affiliations: Department of Clinical Laboratory, Jinling Hospital, State Key Laboratory of Analytical Chemistry for Life Science, Nanjing University School of Life Sciences, Nanjing University, Nanjing, China., State Key Laboratory of Pharmaceutical Biotechnology, Jiangsu Engineering Research Center for MicroRNA Biology and Biotechnology, Nanjing University School of Life Sciences, Nanjing University, Nanjing, China., Department of Urology, Jinling Hospital, Nanjing University School of Medicine, Nanjing University, Nanjing, China.

Abstract:  Although emerging evidence has revealed that microRNAs (miRNAs) dysregulation contribute to carcinogenesis, the mechanism underlying their roles in renal cell carcinoma (RCC) is unclear. The purpose of the current study was to analyze the association of miR-200a-3p expression with RCC and to understand potential novel target genes, functions and mechanisms of miR-200a-3p in RCC. MiR-200a-3p expression levels were first measured by quantitative real-time polymerase chain reaction and in situ hybridization in pairs of RCC tissue samples. Next, the potential miR-200a-3p target gene was analyzed using a combination of computer-aided algorithms, luciferase reporter assays and Western blot analysis. Finally, the biological roles of miR-200a-3p in RCC tumorigenesis were investigated both in vitro by 5-ethynyl-20-deoxyuridine, apoptosis assay and transwell assay, as well as in vivo using a xenograft mouse model. Our results demonstrated that miR-200a-3p was remarkably downregulated in RCC tissues compared with normal adjacent tissue, and CBL is a direct target of miR-200a-3p. An inverse correlation between miR-200a-3p and CBL was observed in RCC tissue samples. Mechanistic investigations revealed that ectopic expression of miR-200a-3p in RCC cell lines suppressed cell proliferation and migration and enforced cell apoptosis by directly inhibiting CBL in vitro and in vivo, whereas silencing miR-200a-3p resulted in the opposite effects. Additionally, overexpressing CBL abolished the effects induced by miR-200a-3p overexpression. Taken together, our results show that the miR-200a-3p/CBL regulation axis is a novel mechanism underlying RCC pathogenesis and may serve as a candidate biomarker and therapeutic target in RCC. 

Retracted: No

PubPeer comments:
  FIGURE 5 MiRu2010200au20103p inhibited RCC cell migration. A, Transwell analysis of ACHN cells treated with miRu2010200au20103p mimics, mimics NC, miRu2010200au20103p inhibitors, and inhibitors NC. B, Transwell analysis of ACHN cells treated with control, CBL plasmid and CBL plasmid plus miRu2010200au20103p mimics . ![file] To overlap one pair of images may be regarded as a misfortune; to overlap both looks like carelessness. , 
 Dear Hoya Camphorifolia, I just learn of the concern that you raised in our paper, and it really astonishes me! We greatly appreciate you can raise this concern for us, and I will contact the first author as well as the co-first author (and also other co-authors) and thoroughly examine the raw data and laboratory records. If we find out the reason, we will contact the journal to explain what has happened and feed back to your website. Thank you very much again for pointing this issue for us! Best personal regards from, Dr. Cheng Wang , 
 Dear Hoya Camphorifolia, Thank you very much for pointing out the errors in the above figures. We have thoroughly examined the raw data and our experiment records, which clearly showed that miR-200a-3p indeed inhibited cell migration by blocking CBL in RCC cells. These errors in the figures were accidently made in the process we elaborated the figures because of our careless. For transwell assay, we generally take one or more representative images of each group for figure presentation. In figure 5A, we wrongly put representative image of miR-NC inhibitor group in the place of miR-NC group, the original data is showed below. For figure 5B, unfortunately, due to the data storage problems, we were unable to find the raw data of the assay showed in the article, but we can present the original data of the repeated assay at the same period (on October 12th 2015), in that assay, CBL showed a strongly enhanced migratory capacity, whereas miRu2010200au20103p could restore the promote effect of CBL, the original data is showed below. Moreover, as we learned the concern that you raised in Figure 5, to further verify the repeatability of our results, we immediately repeated the above assays. The obtained results distinctly confirmed the repeatability of our previous results, and the original data is showed below. Now we have drawn a new figure 5 to correct our errors, which does not affect the interpretation or conclusions of this paper. We really appreciate you for raising this concern for us, and sincerely apologize to our readers for any confusion this unintentional error might have caused. We will also contact the journal to explain what has happened and send correction to address this issue. If you have any question for further clarification, please do not hesitate to contact us (Dr. Ding vikkiding@126.com; Dr. Cheng Wang wangcheng919@smail.nju.edu.cn). Sincerely, Dr. Meng Ding **Correct Figure 5** ![file] FIGURE 5 MiRu2010200au20103p inhibited RCC cell migration. A, Transwell analysis of ACHN cells treated with miRu2010200au20103p mimics, mimics NC, miRu2010200au20103p inhibitors, and inhibitors NC. B, Transwell analysis of ACHN cells treated with control, CBL plasmid and CBL plasmid plus miRu2010200au20103p mimics. ***P 0.001; **P 0.01. miRNA, microRNA; NC, negative control; RCC, renal cell carcinoma. **1. The transwell assay on August 11th 2015** The original data of the transwell assay on August 11th 2015 was stored in the folder named 20150811. Representative images for the four groups of the assay were stored in the subfolders respectively. The subfolder named u201cinhibitoru201d means the group of u201cmiR-200a-3p inhibitoru201d; u201cmimcsu201d refers to u201cmiR-200a-3pu201d; u201cncu201d refers to u201cmiR-NCu201d; u201cnciu201d refers to u201cmiR-NC inhibitoru201d. Three random areas of the same size in each group were finally statistically. ![file] ![file] ![file] ![file] **2. The transwell assay on October 12th 2015** The original data of the transwell assay on October 12th 2015 was stored in the folder named 20151012. Representative images for the three groups of the assay were stored in the subfolders respectively. The subfolder named u201cCBL+200u201d means the group of u201cCBL+miR-200a-3pu201d; u201cCBL+NCu201d refers to u201cCBLu201d; u201cCONTROLu201d refers to u201cControlu201d. Three random areas of the same size in each group were finally statistically. ![file] ![file] ![file] ![file] **3. The confirmation transwell assay on May 25th 2021** The original data of the transwell assay on May 25th 2021 was stored in the folder named 20210525. Representative images named u201cCBLu201d means the group of u201cCBLu201d; u201cCBL+200au201d refers to u201cCBL+miR-200a-3pu201d; u201ccontrolu201d refers to u201cControlu201d; u201cinhibitor 200au201d refers to u201cmiR-200a-3p inhibitoru201d; u201cinhibitor ncu201d refers to u201cmiR-NC inhibitoru201d; u201cmimic 200au201d refers to u201cmiR-200a-3pu201d; u201cncu201d refers to u201cmiR-NCu201d. Three random areas of the same size in each group were finally statistically, the histogram was below, ***P 0.001; **P 0.01. ![file] ![file] ,