Human umbilical cord mesenchymal stem cell-derived exosome-mediated transfer of microRNA-133b boosts trophoblast cell proliferation, migration and invasion in preeclampsia by restricting SGK1
Authors: Dan Wang
Leilei Wang
Quan Na
Gui Yu Song
Journal: Cell Cycle (2020)
Discipline: MOLECULAR BIOLOGY & GENETICS
Keywords: Preeclampsia; SGK1; exosomes; human umbilical cord mesenchymal stem cells; microRNA-133b
Affiliations: Department of Obstetric and Gynecology, Shengjing Hospital of China Medical University , Shengyang, Liaoning, China.

Abstract:  Objective: Exosomes have been documented to function in human diseases, yet their transfer of microRNA (miRNA) in preeclampsia (PE) has seldom been reported. This study intends to discuss the role of miR-133b derived from exosomes in human umbilical cord mesenchymal stem cells (hUC-MSCs) in trophoblast cell development in PE. Methods: Placentas from PE patients and normal pregnant women were collected. The hUC-MSCs and their exosomes were obtained and identified. Trophoblast cell HPT-8 and HTR8-S/Vneo were obtained and co-cultured with hUC-MSCs-derived exosomes that had been transfected with different miR-133b plasmids. MiR-133b and glucocorticoid-regulated kinase 1 (SGK1) expression in placental tissues and HPT-8 and HTR8-S/Vneo cells was determined. HTR8-S/Vneo and HPT-8 cell proliferation, cell cycle distribution, apoptosis rate, migration and invasion were detected. Results: MiR-133b was down-regulated and SGK1 was up-regulated in placental tissues of PE patients. MiR-133b expression was inversely related to SGK1 expression in HTR8-S/Vneo and HPT-8 cells co-cultured with hUC-MSC-derived exosomes. Exosomes promoted HTR8-S/Vneo and HPT-8 cell proliferation, migration and invasion abilities, cell cycle entry and inhibited apoptosis. Elevated exosome-derived miR-133b from hUC-MSCs boosted HTR8-S/Vneo and HPT-8 cell proliferation, cell cycle progression, migration and invasion and limited cell apoptosis. MiR-133b targeted SGK1. Conclusion: Collectively, we demonstrate that miR-133b is down-regulated and SGK1 is up-regulated in PE, and miR-133b derived from exosomes in hUM-MSCs facilitates trophoblast cell proliferation, migration and invasion in PE via constraining SGK1. 

Retracted: No

PubPeer comments:
 Fig 1b.  Alizarin red staining of hUC-MSC osteogenic induction . Boxes around areas of surprising similarity are mine. I am impressed by the skill that went into the preparation of this image; not so much by it being labeled as reality.  , 
 Fig 5.  b. Detection of HTR8-S Vneo cell cycle by flow cytometry... d. Detection of HPT-8 cell cycle by flow cytometry .  These flow-cytometry plots appear to have been hand-drawn. The lines simply do not add up or make sense. , 
 Fig 5.  f. Detection of HTR8-S Vneo cell apoptosis in each group by flow cytometry; ... h. Detection of HPT-8 cell apoptosis in each group by flow cytometry . These scatterplots seem unusual, with their yolk-within-egg structure. Or perhaps they resemble comets.  , 
 Consistent with several other extracellular vesicle (EV) papers highlighted by Hoya Camphorifolia in their tabulation of papers with characteristic features, this paper presents only single panels on EV characterization by NTA, EM, and WB, and confuses EVs with the subpopulation of exosomes. One EV preparation is partially characterized in Figure 1.  Curiously, in addition to the Figure 1d electron micrograph showing no particles with characteristic EV morphology, the  Nanosight panel of Figure 1e only superficially resembles the images generated by genuine NanoSight Malvern software. Unexpected and reality-challenging overhangs are apparent (examples indicated by blue ovals) for the average trace and or the red variability zones, reaching or even exceeding verticality. Were this a standard NTA figure, the legend and text should indicate what the lines and shading represent. The y-axis would also require clarification, since no NTA instrument can detect between 0 and 9 particles per mL of  Conecentration . However, it does not appear that this panel was generated using NTA. The purported flow cytometry in Figure 1f is not explained, nor is the tetraspanin profiling in Figure 1g. However, the authors have apparently solved a vexing problem in the EV field, in that CD63 in Figure 1g behaves as a single band (although size is not indicated) instead of its customary smear. The authors are encouraged to upload full gel images and methods explanations so that others can learn how to tame CD63 glycosylation. , 
 The authors might also wish to issue a correction for Figure 7a, which does not correctly convey the predicted binding of an unspecified miR-133b arm to SGK1.  , 
 It was also unclear from the methods how the mature microRNA was reverse transcribed and amplified. The specified RT kit includes reagents for both poly-A and random hexamer RT, but which was used? And how was the selected method compatible with both miRNA universal primer use and general transcript quantitative PCR? Detailed clarifications of the methods are important to understand how this research was done and how the results should be interpreted. , 
 #4 Hi I think you are right about the smear band of CD63 in human EV samples, but my experience with mouse EVs tells me CD63 can be a single clear band in mouse samples, but not in humans. , 
 Fig 1a. Spotted by ImageTwin.  ,