IL-6 stimulates lncRNA ZEB2-AS1 to aggravate the progression of non-small cell lung cancer through activating STAT1
Authors: J Li
T Chen
L-J Xu
T-C Pan
M-H Zhou
Journal: European review for medical and pharmacological sciences (2020)
Discipline: PHARMACOLOGY & TOXICOLOGY
Keywords: IL-6; non-small cell lung cancer; NSCLC; STAT1; ZEB2-AS1
Affiliations: Department of Cardiovascular Surgery, Tongji Hospital of Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China. .

Abstract:  Objective: To illustrate the role of interleukin 6 (IL-6) in the progression of non-small cell lung cancer (NSCLC) via activating STAT1. Patients and methods: The level of IL-6 mRNA in 48 paired NSCLC tissues and matched normal ones was determined by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). Kaplan-Meier curves were depicted for assessing the overall survival of NSCLC patients with high or low level of IL-6 mRNA. Subsequently, the ZEB2-AS1 level in A549 cells treated with different doses of IL-6 for different time points was determined. After A549 cells were treated with different doses of IL-6, wound closure assays were performed to assess the migration of cells. Protein levels of pSTAT1 and STAT1 in IL-6-treated A549 cells were detected by Western blot. The regulatory effect of STAT1 on IL-6-induced migration of A549 cells was also evaluated. The interaction between ZEB2-AS1 and STAT1 was explored through Chromatin Immunoprecipitation (ChIP) assay. Finally, the role of ZEB2-AS1/STAT1 axis in regulating NSCLC cells was investigated through rescue experiments. Results: Our results indicated that IL-6 was upregulated in NSCLC tissues and cancer cell lines. NSCLC patients with T3-T4 or accompanied with lymphatic metastasis had a higher IL-6 abundance than those with T1-T2 or without metastatic foci. The worse prognosis was identified in NSCLC patients with high expression of IL-6 compared to those with low expression. ZEB2-AS1 showed dose-dependent and time-dependent increase in IL-6-treated A549 cells. IL-6 treatment gradually enhanced the migration ability of A549 cells in a dose-dependent manner. In IL-6-treated A549 cells, protein level of pSTAT1 was remarkably upregulated, and knockdown of STAT1 significantly reversed the promotive effect of IL-6 on migration ability of A549 cells. The results of ChIP assay verified the interaction between ZEB2-AS1 and STAT1. In addition, ZEB2-AS1 could reverse the regulatory effect of STAT1 on the migration ability of A549 cells. Conclusions: IL-6 was upregulated in NSCLC and accelerated the progression of NSCLC via activating STAT1/ ZEB2-AS1. 

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 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? , 
  A total of 48 paired tumor tissues and matched adjacent tissues (3 cm away from the tumor edge) were surgically resected from NSCLC patients treated in Tongji Hospital of Tongji Medical College, Huazhong University of Science and Technology from **April 2016 to December 2018** . Fig 1D,  Kaplan-Meier curves introduced for assessing the overall survival in NSCLC patients with high level or low level of IL-6 . ![file] If I understand correctly, at the time of the paper s publication in 2020, the survival of patients had been followed for 10 years after recruitment, i.e. through to the period from April **2026 to December 2028**. , 
 Here are four Figures 1D, with pairs of 10-year survival curves, to show the range of variation when time-travelling cases of cancer are subgrouped by different biomarkers. All ostensibly different research teams. Clockwise from upper left: - [Chen Wang (2019)] . 48 cases, Non-small-cell lung cancer, ZEB2-AS1. - [Zhou et al (2020)] . NSCLC, ANCR. - **[Chen et al (2020)] . NSCLC, IL-6**. - [Meng et al (2020)] . METTL3. Nasopharangeal cancer, METTL3. ![file] Note the same number of surviving cases in each high-level subgroup when tracking of those cases stopped prematurely after 96 months. ,