|Year : 2016 | Volume
| Issue : 3 | Page : 69-74
High expression of mir-25 predicting poor prognosis in gastric cancer
Chuanli Ren1, Wenshu Wang2, Chongxu Han3, Hui Chen4, Deyuan Fu5, Daxin Wang3, Ming Shen2
1 Department of Clinical Medical Laboratory, Clinical Medical Testing Laboratory, Northern Jiangsu People's Hospital and Clinical Medical College of Yangzhou University, Yangzhou; Department of Epidemiology and Biostatistics, Ministry of Education, Key Laboratory of Modern Toxicology, School of Public Health, Nanjing Medical University, Nanjing, China
2 Department of Chemistry, College of Chemistry and Chemical Engineering, Yangzhou University, Yangzhou, China
3 Department of Clinical Medical Laboratory, Clinical Medical Testing Laboratory, Northern Jiangsu People's Hospital and Clinical Medical College of Yangzhou University, Yangzhou, China
4 Division of Geriatric Medicine, Northern Jiangsu People's Hospital and Clinical Medical College of Yangzhou University, Yangzhou, China
5 Department of Breast Oncology Surgery, Northern Jiangsu People's Hospital and Clinical Medical College of Yangzhou University, Yangzhou, China
|Date of Submission||20-Jul-2016|
|Date of Acceptance||01-Sep-2016|
|Date of Web Publication||30-Sep-2016|
Clinical Medical Testing Laboratory, Northern Jiangsu People's Hospital and Clinical Medical College of Yangzhou University, No. 98 Western Nantong Road, Yangzhou 225001
Source of Support: None, Conflict of Interest: None
Aim: The aim of this study was to explore miR-25 expression pattern and its prognostic value in gastric carcinoma (GC). Methods: MiR-25 expression was detected using miRNA-locked nucleic acid in situ hybridization in 180 patients with GC undergoing surgery. Correlation with clinicopathological features and overall survival (OS) was analyzed. Results: MiR-25 expression was decreased in 10.0% (18/180) of GC, increased in 62.2% (112/180), and unchanged in 27.8% (50/180), compared to samples of morphologically normal tissue taken from the same patient (P < 0.001). Univariate analysis showed that high miR-25 expression, tumor stage, tumor status, node status, and tumor size were significant negative prognostic predictors for OS in patients with GC, and the results were shown as P < 0.001, P < 0.001, P = 0.002, P> 0.001, and P = 0.001, respectively. High miR-25 expression remained a significant predictor of shorter survival in stage II (n = 56, P = 0.015) and stage III (n = 92, P < 0.001) GC. Multivariate regression analysis demonstrated that tumor status (hazard ratio [HR]: 1.91; 95% confidence interval [CI]: 0.79-4.62; P = 0.151), stage (HR: 2.26; 95% CI: 1.30-3.94; P = 0.004), lymph node metastasis (HR: 1.40; 95% CI: 0.73-2.68; P = 0.309), high expression of miR-25 (HR: 2.39; 95% CI: 1.53-3.72; P < 0.001), and tumor size (HR: 2.29; 95% CI: 1.40-3.74; P = 0.001) predicted shorter OS. Conclusion: High expression of miR-25 was associated with decreased OS. Thus, miR-25 may be useful for prognosis evaluation and may provide a novel treatment target in patients with GC.
Keywords: Gastric cancer, miR-25, prognosis, target therapy
|How to cite this article:|
Ren C, Wang W, Han C, Chen H, Fu D, Wang D, Shen M. High expression of mir-25 predicting poor prognosis in gastric cancer. Transl Surg 2016;1:69-74
|How to cite this URL:|
Ren C, Wang W, Han C, Chen H, Fu D, Wang D, Shen M. High expression of mir-25 predicting poor prognosis in gastric cancer. Transl Surg [serial online] 2016 [cited 2019 Sep 22];1:69-74. Available from: http://www.translsurg.com/text.asp?2016/1/3/69/191487
| Introduction|| |
Gastric cancer (GC) is the fifth most common malignant disease globally, with 50% of the total cancer burden occurring in Eastern Asia, predominantly in China.  GC is also associated with tumor relapse and poor overall survival (OS) time. Oncogenic activation (β-catenin) and tumor-suppressor gene (p53) mutations are associated with the histologic and genetic alterations of GC.  The molecular mechanisms depicting the prognosis of GC remain unclear.
Since the discovery of the small RNA molecules, miRNAs have been found to play an important role in regulating cellular biological functions by targeting different genes. ,, In general, miRNAs regulate normal cell function and homeostasis. Moreover, miRNAs are usually an aberrant expression in cancer cells and the tumor microenvironment, and can induce epithelial-to-mesenchymal transition via downregulation of E-cadherin and other proteins. ,, Aberrant miRNA expression in tissue or serum of patients with GC has the potential to be a diagnostic or prognostic biomarker. ,,,,,,,,, However, it is unclear whether these miRNAs are reliable for the role of early diagnosis or prognostic prediction of GC.
MiR-25 is known to be an essential oncogene influencing proliferation and invasion by targeting TOB1,  FBXW7,  and ERBB2.  A single nucleotide polymorphism located on the site of miR-25 (rs41274221) may protect the patients from distant metastasis of GC.  In our previous work, we found that circulating miR-25 was consistently increased in plasma of the GC patients compared with controls, indicating a potential role as a noninvasive biomarker for diagnosis of the early-stage GC.  As the miR-25 expression pattern and its prognostic value are unknown in GC, the goal of this study is to explore the prognostic value of miR-25 in 180 GC patients.
| Methods|| |
Patients and tissue samples
Paraffin-embedded tissue samples were acquired retrospectively from the Biobank Center at the National Engineering Center for Biochip at Shanghai (Shanghai Outdo Biotech Co., Ltd., Shanghai, China). Tumor tissues and corresponding normal tissues were obtained from 180 patients with histologically confirmed GC between 2006 and 2008. Written informed consent was required, and the Ethics Committee of the National Engineering Center approved the protocol for Biochip at Shanghai. OS time was calculated from the date of pathological diagnosis to the time of death or the date of final follow-up. The median follow-up time was 7.1 years, with the range of 6.6-8.1 years [Table 1].
MiRNA-locked nucleic acid in situ hybridization
Tissue microarrays were evaluated using tissue cores from paraffin-embedded GC samples as described by our group and others. ,, MiRNA-locked nucleic acid in situ hybridization was carried out using antisense oligonucleotide probes for miR-25 (Exiqon Inc., Woburn, MA, USA) with a scramble-miRNA serving as a negative control. The method and the definition of the positive and negative miR-25 staining are described in our previous publication. 
Associations between clinicopathological parameters and miR-25 expression were evaluated using Chi-square test. Multivariate Cox proportional hazards regression models were used to analyze the independent prognostic values. All analyses were performed using the SPSS 17.0 statistical software (SPSS Inc., Chicago, IL, USA). All tests were two-sided and P < 0.05 was considered statistically significant.
| Results|| |
Expression of miR-25 in gastric carcinoma
MiR-25 was mainly expressed in the cytoplasm of cells of GC and normal gastric tissue demonstrated by in situ hybridization [Figure 1]. MiR-25 expression was increased in 62.2% (112/180), decreased in 10.0% (18/180) of GC, and unchanged in 27.8% (50/180), compared to normal tissue (P < 0.001).
|Figure 1. MiR-25 levels stained using in situ hybridization. (a) MiR-25 staining using in situ hybridization in gastric carcinoma. (b) MiR-25 staining in adjacent normal tissue; increased miR-25 expression in gastric carcinoma compared with neighboring normal tissue. (c) MiR-25 staining using in situ hybridization in gastric carcinoma. (d) MiR-25 staining in adjacent normal tissue; increased miR-25 expression in gastric carcinoma compared to adjacent normal tissue|
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MiR-25 expression and its clinicopathological features in gastric carcinoma
No significant correlations were found between miR-25 expression levels and other clinicopathological variables, including age (P = 0.891), gender (P = 0.760), tumor site (P = 0.938), tumor size (P = 0.185), nodal status (P = 0.287), TNM stage (P = 0.166), local invasion (P = 0.185), and distant metastasis (P = 0.151) [Table 2].
|Table 2: MiR - 25 expression and clinicopathological features in patients with gastric adenocarcinoma|
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Kaplan-Meier analysis indicated that high expression of miR-25 (P < 0.001), stage of disease (P < 0.001), tumor status (P = 0.008), node status (P < 0.001), tumor size (P = 0.001), and distant metastasis (P = 0.005) were related to significantly shorter OS in patients with GC. Age, gender, and location did not correlate with OS (P > 0.05) [Table 3].
|Table 3: Univariate analysis of survival in patients with gastric cancer|
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High expression of miR-25 (n = 112) remained a significant predictor of shorter mean OS compared with low/unchanged group (n = 68) (43.1 vs. 66.7 months, P < 0.001) [Figure 2]. After TNM stratification, high expression of miR-25 remained a significant predictor of poor OS in stage II (47.0 vs. 61.5 months, P = 0.0015, n = 56) and stage III GC (36.3 vs. 67.2 months, P < 0.001, n = 92) [Figure 2].
|Figure 2. Survival curves in patients with gastric carcinoma according to miR-25 levels. (a) Overall survival curves in 180 patients with gastric carcinoma according to miR-25 levels (P < 0.001). (b) Survival curves of patients with stage II gastric carcinoma according to miR-25 levels (P = 0.015). (c) Survival curves of patients with stage III gastric carcinoma according to miR-25 levels (P < 0.001)|
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Univariate Cox regression analysis showed predictors of poor prognosis as following: tumor status (hazard ratio [HR]: 3.10; 95% confidence interval [CI]: 1.51-6.37; P = 0.002), stage (HR: 2.83; 95% CI: 1.88-4.27; P < 0.001), lymph node metastasis (HR: 2.82; 95% CI: 1.70-4.68; P < 0.001), high miR-25 expression (HR: 2.53; 95% CI: 1.64-3.90; P < 0.001), and tumor size (HR: 2.30; 95% CI: 1.42-3.71; P = 0.001). Gender, age, and tumor site were not predictors of poor prognosis [Table 4].
|Table 4: Multivariate Cox regression analysis of potential survival prognostic factors in 180 patients with gastric cancer|
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Multivariate Cox regression analysis identified predictors of poor prognosis as following: tumor stage (HR: 2.26; 95% CI: 1.30-3.94; P = 0.004), miR-25 (HR: 2.39; 95% CI: 1.53-3.72; P < 0.001), age (HR: 1.21; 95% CI: 0.80-1.82; P = 0.372), and tumor site (HR: 1.82; 95% CI: 1.30-3.94; P = 0.03). Gender, tumor status, lymph node metastasis, tumor site, and tumor size were not predictors of poor prognosis [Table 4].
| Discussion|| |
To date, p21,  ERBB2,  FBXW7, , RhoGDI1,  SOCS4,  RGS3,  NEFL,  MOAP1,  CDKN1C,  KLF4, C/EBPα,  αv- and α6-integrins  have been reported as potential targets for miR-25 in various solid cancers. MiR-25 may act as an oncogenic miRNA to promote proliferation, migration, and invasion in many kinds of cancers, such as GC, ,,, colorectal cancer, , lung cancer, ,,,, ovarian cancer,  and esophageal squamous cell carcinoma.  However, miR-25 polymorphism can inhibit oncogenic function in GC. 
Circulating miR-25 in the plasma/serum could be used as a noninvasive biomarker for the early diagnosis of GC,  hepatocellular carcinoma,  and lung cancer.  According to recent publications, circulating miR-25 has diagnostic and prognostic value in both ovarian and hepatocellular cancer patients. ,
In our previous work, we found that circulating miR-25 was part of a five-miRNA signature for early diagnosis of GC.  Some groups have shown that high expression of miR-25 can be used as a predictive prognostic marker in hepatocellular carcinoma.  We compared miR-25 expression levels in GC and surrounding morphologically normal tissues and found that miR-25 expression was significantly increased in GC tissues overall. However, its expression level was decreased in 10.0% (18/180) of samples and unchanged in 27.8% (50/180), suggesting that heterogeneous miR-25 expression may have different prognostic values in different individuals.
To the best of our knowledge, this study is the first to systematically study the relationship of miR-25 to clinicopathological parameters and prognosis in patients with GC. In our previous work, we identified increased circulating miR-25 expression in the early stages of GC.  In addition, Li et al. found that high plasma concentrations of miR-25 in GC patients predicted poor prognosis.  This implies that higher levels of circulating and tissue miR-25 in GC may predict poor prognosis.
In this study, GC patients with higher miR-25 had a worse prognosis than those with lower or unchanged expression. Moreover, using multivariate Cox regression analysis, we identified high levels of miR-25 indicating poor prognosis (P < 0.001). In a univariate Cox regression analysis, high miR-25 expression was also independently associated with a worse prognosis (P < 0.001). Li et al. also found that circulating miR-25 in GC patients correlated with poor prognosis. As expected, the elevated level of miR-25 in plasma and GC tissues was consistent. Therefore, increased circulating miR-25 in the plasma may originate primarily from GC cells. In this study, we found that miR-25 was sometimes overexpressed in normal surrounding gastric tissues and its fundamental function is unknown. Thus, detection of miR-25 in the serum or tissues may provide new information to help early diagnosis and prognostic evaluation of GC. Although the molecular function of miR-25 in GC is largely unknown, miR-25 may act as an oncogenic driver in GC. ,,,
Although intervention including surgical resection may be different for each patient, higher expression of miR-25 remained a poor prognosticator in dependent of other clinical parameters. Other clinical and genetic factors may influence OS. For example, groups with a high expression of miR-25 trended toward more advanced TNM stage at presentation though this was not statistically significant [Table 2]. There are other factors that may influence tumor biology, such as p53 polymorphism, that could offset the efficacy of chemotherapy and affect the OS. , The molecular mechanism of increased miR-25 and its relationship to diagnosis and prognosis requires further investigation and validation.
| Conclusion|| |
Our study indicates that in patients with GC, increased expression of miR-25 predicts a relatively poor prognosis while low/unchanged miR-25 expression indicates a better prognosis. Although further research is needed to confirm our findings, this study suggests that miR-25 may represent a potential biomarker and target for the treatment of GC.
Financial support and sponsorship
This study was supported by the National Nature Science Foundation of China (Grant Number: 81573220), the Jiangsu provincial Six Talent Peaks (Grant Number: WSN107), the Foundation of China Postdoctoral Studies (Grant Number: M2013541699), the Foundation of Jiangsu Province Postdoctoral Studies (Grant Number: 1302149C), and the Foundation for Yong Scholar in Yangzhou (Grant Number: YZ2014046).
Conflicts of interest
There are no conflicts of interest.
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[Figure 1], [Figure 2]
[Table 1], [Table 2], [Table 3], [Table 4]