miR-34b/c rs4938723 T>C Decreases Neuroblastoma Risk: A Replication Study in the Hunan Children

Hindawi Disease Markers Volume 2019, Article ID 6514608, 6 pages https://doi.org/10.1155/2019/6514608 Research Article miR-34b/c rs4938723 T>C Decreases Neuroblastoma Risk: A Replication Study in the Hunan Children Yong Li,1 Zhen-Jian Zhuo,2 Haiyan Zhou,3 Jiabin Liu ,2 Zhenghui Xiao,4 Yaling Xiao,1 Jing He ,2 and Zan Liu 1 1 Department of Pediatric Surgery, Hunan Children’s Hospital, Changsha, 410004 Hunan, China Department of Pediatric Surgery, Guangzhou Institute of Pediatrics, Guangzhou Women and Children’s Medical Center, Guangzhou Medical University, Guangzhou, 510623 Guangdong, China 3 Department of Pathology, Xiang-ya School of Medicine, Central South University, Changsha, 410013 Hunan, China 4 Emergency Center of Hunan Children’s Hospital, Changsha, 410004 Hunan, China 2 Correspondence should be addressed to Jing He; and Zan Liu; Received 3 May 2019; Revised 30 July 2019; Accepted 13 August 2019; Published 10 September 2019 Academic Editor: Kishore Chaudhry Copyright © 2019 Yong Li et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Neuroblastoma is the most common seen solid neural tumor in children less than age one. As mutation in the miR-34b/c gene is observed in several types of human malignancies, there likely to be similar events that contribute to the pathogenesis of neuroblastoma. We hypothesize that polymorphism in the miR-34b/c gene might predispose to neuroblastoma. Here, we conducted this replication study by genotyping rs4938723 T>C from miR-34b/c in Hunan children (162 subjects with neuroblastoma and 270 control subjects) and examined its effect on the risk of neuroblastoma. We determined such association using logistic regression, adjusted for age and gender. Relative to those with TT genotype, subjects with C allele had reduced neuroblastoma risk (TC vs. TT: adjusted OR = 0 46, 95%CI = 0 30‐0 71; additive model: adjusted OR = 0 64, 95%CI = 0 47‐0 88; TC/CC vs. TT: adjusted OR = 0 49, 95%CI = 0 33‐0 73). Stratified analysis revealed that rs4938723 TC/CC carriers were less likely to develop neuroblastoma for patients in the subgroups of age ≤ 18 months, age > 18 months, females, males, tumors in retroperitoneal, tumors in other sites, and clinical stages II, III, IV, and III+IV. Our findings verified miR-34b/c rs4938723 C variant allele as a protective factor for the risk of neuroblastoma. Further investigation of how miR-34b/c rs4938723 T>C might modify neuroblastoma risk is warranted. 1. Introduction Neuroblastoma is a childhood tumor that mainly derives from neural crest progenitor cells [1–3]. Despite representing about 8-10% of all pediatric cancer diagnoses, neuroblastoma disproportionately results in 12-15% of all childhood cancer-related mortality [4–6]. It is characterized by widely clinical heterogeneity, spans from spontaneous regression to therapy-refractory progression [7]. Another reflection of such heterogeneity was the contrasting survival rate of different subgroup patients [8, 9]. In patients with the lowand intermediate-risk neuroblastoma, the long-term survival rate is greater than 90% [10]. However, in patients with the high-risk neuroblastoma, less than 40% could finally survive [11, 12]. In the past decades, considerable progress has been made in understanding the genetic underpinnings of neuroblastoma. Exposed environmental factors of children and pregnant women were reported to predispose to neuroblastoma, but not finally defined [13, 14]. Mutations in ALK [15] and PHOX2B [16] were considered as two major causes of familial neuroblastoma. Other SNPs in genes including LMO1 [17], BARD1 [18], TP53 [19], LIN28B [20], HACE1 [20], NEFL [21], and CDKN1B [22] have more recently been identified to be associated with neuroblastoma predisposition. Moreover, the association of these SNPs to neuroblastoma risk has also been replicated in many other populations, especially the SNPs in the BARD1 gene [23–25]. Taken together, however, all the current identified mutations still could not fully elucidate the etiology of neuroblastoma. We are still 2 on the way to fully reveal the genetic landscape of neuroblastoma. Identification of other somatic mutations will further clarify the mechanisms of neuroblastoma. MicroRNAs (miRNAs) are a class of nonprotein-coding, small, single-stranded RNAs with about 22 nucleotides [26]. miRNAs participate in transcriptional regulation through multiple mechanisms, including mRNA degradation, translational repression, or cleavage of mRNA [26–28]. In the past decade, more and more miRNAs are being identified that play vital regulatory roles in human disorders, including cancers. Mutations or single nucleotide polymorphisms (SNPs) in miRNA genes may alter the binding ability of miRNAs to their target mRNAs, thus resulting in diverse functional consequences and thereby possibly impact cancer susceptibility [29, 30]. rs4938723 T>C is located at the promoter region of pri-miR-34b/c [31]. Such T to C shift polymorphism might cause a disruption of GATA-X transcription factor binding capacity, which results in decreased pri-miR34b/c expression [32]. Thus far, most studies have addressed the identification of miR-34b/c rs4938723 T>C in breast cancer [33], colorectal cancer [34], hepatocellular cancer [35], and nasopharyngeal carcinoma [36], whereas few studies focused on the role of miR-34b/c gene rs4938723 T>C in neuroblastoma risk. In our previous study conducted recently, we firstly found that rs4938723 T>C polymorphism was associated with a significantly decreased neuroblastoma risk [37]. Here, we further conducted a replication hospitalbased case-control study aiming to verify the association between miR-34b/c rs4938723 T>C and neuroblastoma risk in Hunan children. 2. Materials and Methods 2.1. Study Subjects. Prior to analysis, the study protocols were approved by the Institutional Review Board of Hunan Children’s Hospital. The current case-control study was carried out in Hunan Children’s Hospital. A total of 162 cases were pathology-confirmed with neuroblastoma, and 270 controls with no prior history of neuroblastoma were randomly enrolled in the same area as cases. All guardians of participants provided written informed consent. The detailed information of selection criteria of study subjects was reported in our previous paper [38–40]. 2.2. Genotyping. Genomic DNA was isolated from venous blood using a TIANamp Blood DNA Kit (TianGen Biotech Co. Ltd., Beijing, China). Genotype analysis of miR-34b/c gene rs4938723 T>C was undertaken using TaqMan SNP genotyping assay from Applied Biosystems [41–44]. Negative controls (with water) and duplicate test samples (10% of all the samples) were included in each 384-well plate. 100% concordant of genotypes in replicates were achieved. 2.3. Statistical Analysis. Tests for the Hardy-Weinberg equilibrium (HWE) (...truncated)