miR-125a restrains cell migration and invasion by targeting STAT3 in gastric cancer cells
Received 16 March 2018
Accepted for publication 18 November 2018
Published 24 December 2018 Volume 2019:12 Pages 205—215
Checked for plagiarism Yes
Review by Single-blind
Peer reviewers approved by Dr Amy Norman
Peer reviewer comments 5
Editor who approved publication: Prof. Dr. Geoffrey Pietersz
Liu Yang,1,* Shuguang Zhang,2,* Kai Guo,3,* Hu Huang,4,* Shuai Qi,5 Jie Yao,6 Zhihong Zhang7
1Department of Cancer Biotherapy Center, Hubei Cancer Hospital, Wuhan, Hubei 430079, China; 2Department of Clinical Laboratory, Liaocheng People’s Hospital, Liaocheng, Shandong 252000, China; 3Department of Gastroenterology, The 161th Hospital of PLA, Wuhan, Hubei 430010, China; 4Department of Oncology, The 161th Hospital of PLA, Wuhan, Hubei 430010, China; 5Department of Pharmacy, The 161th Hospital of PLA, Wuhan, Hubei 430010, China; 6Department of Urological Surgery, Zhongnan Hospital of Wuhan University, Wuhan, Hubei 430071, China; 7Department of Oncology, Gong’an County People’s Hospital, Jingzhou, Hubei 434000, China
*These authors contributed equally to this work
Background: Recently, many microRNAs have been found to be involved in the cancer progression including miR-125a. However, the underlying mechanisms of miR-125a in gastric cancer (GC) remain to be completely elucidated.
Objective: The study was to investigate the functional role of miR-125a and the expression relevance of signal transducer and activator of transcription 3 (STAT3) and hyaluronan synthase 1 (HAS1).
Method: CCK-8 assay, scratch wound healing and transwell assay were conducted to identify the functional role of miR-125a in GC. In addition, using bioinformatics analysis, the target regulation relationship was found in STAT3 and miR-125a. To confirm the relationship, luciferase reporter assay was performed. More importantly, quantitative polymerase chain reaction and western blot assay were carried out to determine the association among miR-125a, STAT3 and HAS1 in GC cells.
Results: Overexpressed miR-125a inhibited the migration and invasion of GC cells through scratch wound healing and transwell assay, and its knockdown displayed adverse effects, but the viability of GC cells did not show significant difference using CCK-8 assay. In addition, we identified that the knockdown of STAT3 or HAS1 remarkably suppressed the migration and invasion abilities of GC cells. Using bioinformatics analysis, miRTar, in particular, indicated that the 3'-untranslated region of STAT3 binds to miR-125a with a high score. Subsequently, we also verified that STAT3 was a target of miR-125a via luciferase reporter assay. Furthermore, we found that upregulated miR-125a expression could conspicuously constrain STAT3 expression at both protein and mRNA levels in MKN45 and NCI-N87 cells using quantitative polymerase chain reaction and Western blot assay, but no significant difference had been found in SGC 7901 cells. To further identify the regulatory relationship between miR-125a and STAT3, downregulation of miR-125a in MKN45 and NCI-N87 cells was carried out, which showed that the protein and mRNA expression levels of STAT3 were declined in two cell lines. Finally, we observed that upregulated miR-125a could lead to the decrease of HAS1 at protein and mRNA levels, whereas its knockdown revealed opposite effects. Meanwhile, we noticed that overexpression of STAT3 could induce the escalation of HAS1 at protein and mRNA expression levels and its knockdown exhibited the adverse outcomes.
Conclusion: These findings indicated that miR-125a may control the HAS1 expression in GC progression by targeting STAT3, which is likely to facilitate a better understanding of the regulation mechanisms of miR-125a in GC.
Keywords: miR-125a, STAT3, HAS1, gastric cancer
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