Expression of microRNA-328 Functions as a Biomarker for Recurrence of Early Gastric Cancer (EGC) After Endoscopic Submucosal Dissection (ESD) by Modulating CD44

Abstract

BackgroundThis study investigated the molecular mechanism of the effect of CD44 on the recurrence of EGC after ESD, including the potential regulator and signaling pathways of CD44.Material/MethodsWe searched the miRNA online database (www.mirdb.org) with the “seed sequence” located within the 3′-UTR of the target gene, and performed luciferase assay to test the miRNA/mRNA relationship. We also determined the expression of CD44 in the EGC and control samples. In addition, statistical analysis was used to explore the role of miR-328 as a biomarker to predict the recurrence after ECD.ResultsWe validated CD44 to be the direct gene via luciferase reporter assay system. We also established the negative regulatory relationship between miR-328 and CD44 via studying the relative luciferase activity at different concentrations of miR-328 mimics. We also conducted real-time PCR and Western blot analysis to study the mRNA and protein expression level of CD44 among different groups (recurrence-positive and recurrence-negative) or cells treated with different concentrations of miR-328 mimics/inhibitors, indicating the negative regulatory relationship between miR-328 and CD44. We also investigated the relative viability of EGC cells when transfected with miR-328 mimics (50 nM and 100 nM) and miR-328 inhibitors (100 nM) to validate miR-328 to be negatively interfering with the viability of EGC cells. miR-328 was also recognized as a potential biomarker to predict recurrence after ESD in EGC patients via analysis of the recurrence-free rate among different groups of EGC patients.ConclusionsThe expression level of miR-328 can function as a predictive biomarker of recurrence after ECD in patients with EGC via targeting CD44.

Study on miRNAs in Pan-Cancer of the Digestive Tract Based on the Illumina HiSeq System Data Sequencing

“…[29–33]. This is similar to the result of Zhang et al [27], who studied the correlation between five miRNAs (miR-191, miR-28-3p, miR-145, miR-328, and miR-18a) in the serum of NSCLC patients and the three-year overall survival rate of the two groups. The result showed that miR-328 was differentially expressed in paired tumors and normal tissues ( P < 0.0001), that the target gene of miR-328 was mainly enriched in inflammatory/immune response pathways and pathways related to chemoradiotherapy and/or targeted treatment resistance, and that the miR-328 signal could play a role as a prognostic factor for patients with advanced NSCLC [29, 34].…”

“…On the contrary, Xue et al discussed the role of miR-328 as a biomarker to predict tumor recurrence by studying the luciferase activity in the simulant of miR-328 with different concentrations. It was found that miR-328 could target CD44, and they showed a negative regulatory relationship between them, which indicated that detecting the expression level of miR-328 can be used as a means to predict the prognosis of gastric cancer patients [27]. miR-328 has been found to be associated with the prognosis of a variety of tumors in previous studies.…”

CD44 as a tumor biomarker and therapeutic target

“…On the other hand, CD44 also can be regulated by various molecules, signaling and some drugs. Some micro-RNAs have been found to negatively modulate CD44 expression, such as microRNA-150 [39], microRNA-200c [88], microRNA-34a [33], microRNA-330-5p [89], micro-RNA-145 [90], microRNA-3129 [91], microRNA-143 [92], microRNA-328 [93], and microRNA-373 [94]. CD44 has been reported to be a novel downstream target of microRNA-150 in nasopharyngeal carcinoma [39].…”

Informatics Inference of Exercise-Induced Modulation of Brain Pathways Based on Cerebrospinal Fluid Micro-RNAs in Myalgic Encephalomyelitis/Chronic Fatigue Syndrome

“…miR-200a-5p modulated PTEN, which recycles inositol-trisphosphate for the PIK3R3-AKT pathway ( Table 5). [49][50][51][52][53][54][55][56][57][58][59][60][61][62][63][64][65] IPA MicroRNA Target Filter. The five miRNAs were searched as a group with IPA MicroRNA Target Filter tool.…”