Upregulation of miR-494 Inhibits Cell Growth and Invasion and Induces Cell Apoptosis by Targeting Cleft Lip and Palate Transmembrane 1-Like in Esophageal Squamous Cell Carcinoma
Authors: Wenqiao Zang
Guoqiang Zhao
Yuwen Du
Tong Li
Shengjie Zhang
Ren Zhang
Xiaonan Chen
Zhicai Liu
Jinwu Wang
Xueyan Zhang
Yuanyuan Wang
Journal: Digestive Diseases and Sciences (2015)
Discipline: CLINICAL MEDICINE
Keywords: miR-494; esophageal squamous cell carcinoma (ESCC); CLPTM1L; bioinformatics analyses; qRT-PCR assays
Affiliations: College of Basic Medical Sciences, Zhengzhou University, No. 100 Kexue Road, Zhengzhou, 450001, Henan, China.

Abstract:  Background: Potential target genes of microRNA (miR)-494 have been reported in many types of cancers. However, the role of miR-494 in esophageal squamous cell carcinoma (ESCC) remains unknown. Aim: This study focused on the expression and biological function of miR-494 in ESCC. Methods: Using bioinformatics analyses, we found that cleft lip and palate transmembrane 1-like (CLPTM1L) was a potential target of miR-494. We performed quantitative real-time (qRT) PCR assays in 37 ESCC tumor tissues to determine the expression of miR-494 and CLPTM1L mRNA, and we analyzed the correlation between both of these factors and clinical characteristics. The cell counting kit-8 and colony formation assays were used to evaluate the effects of miR-494 expression on the proliferation of ESCC cells. The transwell migration assay and flow cytometric apoptosis assay were performed to study the influence of miR-494 on the invasion and apoptosis of ESCC cells. Western blotting, luciferase assays, and CLPTM1L knockdown experiments were used to determine whether CLPTM1L was a target of miR-494. Results: The qRT-PCR assays showed significant downregulation of miR-494 (P &lt; 0.05) and upregulation of CLPTM1L mRNA (P &lt; 0.05), both of which were significantly associated with lymph node metastases (P &lt; 0.05). High expression of miR-494 inhibited cell proliferation and invasion and promoted cell apoptosis (P &lt; 0.05). The results also showed that CLPTM1L was a target of miR-494. Conclusion: These results show that the expression of miR-494, which can regulate cell growth, invasion and apoptosis of ESCC cells by targeting CLPTM1L, is downregulated in ESCC tumor tissues. The miR-494-CLPTM1L pathway could be further exploited to develop a new approach to treat ESCC. 

Retracted: No

PubPeer comments:
 From the methods described below, it appears that patients consent ethical board approval were not required obtained. Could the authors provide these documents? Patients and controls Full thickness specimens from the non-inflamed proximal resection margin of 27 patients (12 men and 15 women; age range, 32u201387 years) undergoing left hemicolectomy for colonic diverticulitis were retrieved from our archival pathology laboratory. Two groups of patients were evaluated: 12 patients (eight men and four women, aged 59u2009u00b1u200913 years) undergoing emergency surgery for purulent faecal peritonitis resulting from free perforation of a diverticulum (Hinchey stages IIIu2013IV [18], severe disease according to Ambrosetti classification [19]) and 15 patients (4 men and 11 women, aged 64u2009u00b1u200912 years) undergoing elective surgery after the third or fourth attack of diverticulitis [20, 21]. Control samples were obtained from ten patients (five women and five men; age range, 41u201378 years) undergoing left hemicolectomy for non-obstructing cancer. These patients were not constipated and had no colonic dilatation. Control specimens were taken at least 3 cm from the resection margin in tumour-free areas. Ethical considerations Since this was a retrospective study, no individual patient identification was involved and no study-driven clinical intervention was performed; therefore, no ethical approval was necess , 
 The square highlight the portion that has been reproduced in the two papers. Could the authors explain.? ![file] ,