microRNA‐377 acts as a suppressor in esophageal squamous cell carcinoma through CBX3‐dependent P53/P21 pathway

Abstract Stem cells play pivotal roles in esophageal squamous cell carcinoma (ESCC) recurrence and metastasis. The self‐renewal ability of stem cells was associated with specific microRNAs (miRs). Herein, we identified the effects of miR‐377 on ESCC stem cell activities. First, the expression of miR‐377 in ESCC and adjacent normal tissues was determined. The relationship between miR‐377 and chromobox protein homolog 3 (CBX3) was assessed by a dual‐luciferase reporter gene assay. miR‐377 was overexpressed or inhibited in ESCC stem cells to explore its role in ESCC. To further investigate the mechanism of miR‐377 in ESCC, cells were introduced with short hairpin RNA against CBX3 or pifithrin‐α (inhibitor of P53 pathway). Besides, the expression of P21, P53, CD133, CD13, Nanog, sex determining region Y‐Box 2 (Sox2), and octamer‐binding transcription factor 4 (Oct4), cell sphere formation, colony formation, and proliferation were evaluated respectively. Finally, limiting dilution assay in vivo and tumor xenograft in nude mice were conducted to confirm the roles of miR‐377 in vivo. miR‐377 was poorly expressed in ESCC. Overexpression of miR‐377 could suppress the stem‐like trait of ESCC as well as the tumor growth in vivo. miR‐377 targeted CBX3 to activate the P53/P21 pathway. Besides, the expression of stem‐like markers including CD133, CD13, Oct4, Sox2, and Nanog was decreased, and the abilities of cell sphere formation, colony formation, proliferation, and tumorigenicity were significantly reduced by overexpressing miR‐377 or silencing CBX3. The results were reversed after inactivating the P53/P21 pathway. In summary, upregulation of miR‐377 inhibits the self‐renewal of ESCC stem cells by inhibiting CBX3 expression and promoting activation of the P53/P21 pathway.