Midazolam inhibits proliferation and accelerates apoptosis of hepatocellular carcinoma cells by elevating microRNA‐124‐3p and suppressing PIM‐1
Authors: Xianhui Du
Xiangyan Yao
Yanyan Qi
Journal: IUBMB Life (2020)
Discipline: BIOLOGY & BIOCHEMISTRY
Keywords: HepG2 cell; PIM-1; hepatocellular carcinoma; malignant phenotype; microRNA-124-3p
Affiliations: Anesthesiology Department, Henan Province People's hospital, Zhengzhou, Henan, People's Republic of China.

Abstract:  Objective: Recently, the impact of microRNAs (miRNAs) has been identified in hepatocellular carcinoma (HCC), this study was designed to assess the effects of miR-124-3p and midazolam (MDZ) in HCC with the involvement of PIM-1. Methods: HepG2 human HCC cells were selected for our study, which were treated with different concentrations of MDZ. The gain- and loss-of-function experiments were performed to elucidate the migration, invasion, proliferation, colony formation ability, cell cycle, and apoptosis of HepG2 cells upon treatment of MDZ, miR-124-3p mimics, or miR-124-3p inhibitor. The expression levels of miR-124-3p, PIM-1, Bax, Bcl-2, P21, and Ki-67 in HepG2 cells were assessed by reverse transcription quantitative polymerase chain reaction and western blot analysis. Moreover, HepG2 cell growth in vivo was measured by subcutaneous tumorigenesis in nude mice, and the target relation between miR-124-3p and PIM-1 was evaluated using dual luciferase reporter gene assay. Results: We have found that after treated with overexpression of miR-124-3p and MDZ, there exhibited elevated miR-124-3p, declined expression of PIM-1, attenuated migration, invasion, proliferation and colony formation ability, and promoted apoptosis of HepG2 cells. Additionally, it could be observed that the tumor volume and weight were all reduced upon treatment of overexpression of miR-124-3p and MDZ. Meanwhile, the results in the HepG2 cells that treated with down-regulated miR-124-3p were the opposite. Furthermore, PIM-1 was found to be a target gene of miR-124-3p. Conclusion: Our study found that MDZ could inhibit proliferation and accelerate apoptosis of HCC cells by elevation of miR-124-3p and suppressing PIM-1, which may be an effective method in the treatment of HCC. 

Retracted: No

PubPeer comments:
 Figure 1. Some cells visible in the M-MDZ panels appear to also be visible in the H-MDZ panels but in a different configuration. In addition, the bottom panels appear to contain repetitive elements. Boxes of the same color bring attention to groups of cells that look similar. ![file] , 
 Figure 3E: The fifth panel from the left (and possibly also the fourth) appears to have some unexpected repetitive elements. Highlighted with pink and turquoise boxes. ![file] , 
 Figure 4C panels appear to show some repetitive elements, either within or between panels. Boxes of the same color draw the reader s attention to some of those similar-looking areas. ![file] , 
 Figure 5B shows  Representative pictures for tumors in each group 25 days after the injection with three nude mice in each group. Surprisingly, some of these tumors look more similar to each other than expected, albeit with some rotation or mirroring. Boxes of the same color draw the reader s attention to tumors that share similarities. ![file] , 
 Finally, I have some questions about the authors contributions. The article states the following contributions: * Y.Q. guarantor of integrity of the entire study. * X.D. designed the study. * X.Y. literature research. My questions are: 1. Who actually performed the experiments? The paper describes a range of experiments including cell transfections, microscopy, flow cytometry, RT-PCR, Western blots, and mouse experiments. 2. What is meant by  guarantor of integrity ? That is not a usual role that warrants authorship. 3. Who wrote the paper? , 
 #4 ![file] , 
 Fig 4A. What happened to you, Flow Cytometry cell-cycle plots? ![file] Who did this to you?! ![file] , 
 Fig 2a.  The effects of MDZ and miR-124-3p on invasion and migration of HepG2 cells were detected by Transwell assay . It was good of Dr Bik to leave this Figure for other people to play with. ![] ,