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Journal of Gastrointestinal Cancer

, Volume 46, Issue 3, pp 251–258 | Cite as

Downregulation of the Genes Involved in Reprogramming (SOX2, c-MYC, miR-302, miR-145, and P21) in Gastric Adenocarcinoma

  • Mitra Khalili
  • Mohammad Vasei
  • Davood Khalili
  • Kamran Alimoghaddam
  • Majid Sadeghizadeh
  • Seyed Javad MowlaEmail author
Original Research

Abstract

Purpose

Many cell signaling pathways essential for normal stem cell development are involved in cancer initiation and progression. In the present study, motivated by a possible contribution of reprogramming process in induction of cancer, we compared the expression level of main genes involved in iPS generation, i.e., miR-302, miR-145, SOX2, c-MYC, and P21, in a series of tumor and non-tumor tissues of stomach.

Methods

A total number of 34 tumors and their matched non-tumor (as control) gastric surgical specimens were obtained. The expression of the candidate genes was evaluated by using real-time PCR and immunohistochemistry (IHC) techniques.

Results

Our data revealed a significant downregulation of miR-302b, P21, and miR-145 genes in intestinal and SOX2 gene in diffuse type of tumor samples. SOX2, but not the other genes, showed a significant downregulation in both proximal (cardia and fundus) and distal (body and antrum) sites of stomach. Based on receiver-operating characteristic (ROC) analyses, the highest total area under the curve (AUC) was found for SOX2 (AUC = 82 %, P < 0.001). Interestingly, all tumor samples revealed a negative signal for c-MYC expression, while non-tumor samples represented an intense cytoplasmic staining.

Conclusions

Despite the fact that some hESC-specific genes are upregulated in tumors, our data revealed a significant downregulation of all candidate genes, except for c-MYC, in tumor samples of stomach. Moreover, ROC data demonstrated that SOX2 gene expression index is a better potential biomarker of gastric cancer, compared to other tested genes. SOX2 expression has a good sensitivity and specificity to discriminate correctly between tumor/non-tumor and also high/low grades of tumor malignancy. It seems downregulation of miR-302b, miR-145, and P21 could contribute to gastric tumor initiation and progression.

Keywords

Reprogramming Cancer stem cell SOX2 miR-302b miR-145 P21 Gastric cancer 

Notes

Acknowledgments

We are grateful to Dr. Forouzandeh Fereidooni, the previous head of the Iran Tumor Bank, Dr. Zahra Hosseini, Dr. Fatemeh Kamali, and Mr. Ahmad Joulaie for supplying clinical samples and providing patients’ clinicopathological information. All clinical samples were provided by Iran National Tumor Bank (which is funded by the Cancer Institute of Tehran University, for cancer research). We also offer special appreciation to Prof. Reza Malekzadeh (Tehran University of Medical Sciences) for worthwhile advisors and Mrs. Rozita Edalat, Mehdi Parniyan and Vahid Kia (Pasteur Institute of Iran) for providing technical advice concerning Real-Time PCR.

Conflict of Interest

The authors have no conflict of interests.

Supplementary material

12029_2015_9695_Fig6_ESM.jpg (91 kb)
Online Resource 1

Comparative gene expression in different sites of stomach. Histograms show the median value of relative gene expression in tumor and non-tumor samples in proximal (Cardia and Fudus) and distal (Body and Antrum) sites of stomach. A significant down-regulation in both sites was exclusively observed for SOX2 gene, whereas miR-302b, P21 and miR-145 showed significant differences only in the proximal site of stomach (p-value <0.01 based on Wilcoxon test). * According to Dunn-Bonferroni correction method a threshold value of 0.01 was considered as statistically significant. (JPEG 91.2 kb)

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12029_2015_9695_Fig7_ESM.gif (10 kb)
Online Resource 2

Correlation between the expressions of candidate genes. The correlation coefficients of all genes were analyzed by using the Spearman’s rho test. According to this test, miR-302b showed a significant correlation with miR-145. (*) and (**) signs indicate that the correlation is significant at the 0.05 and 0.001 levels (2-tailed) respectively. (GIF 10.2 kb)

12029_2015_9695_MOESM2_ESM.tif (740 kb)
High Resolution (TIFF 740 kb)
12029_2015_9695_Fig8_ESM.gif (44 kb)
Online Resource 3

Immunocytochemistry for SOX2 protein expression in NT2 cell line. Activity of SOX2 primary antibody was confirmed by immunocytochemistry on human embryonic carcinoma cell line NT2, as a positive control. Antibody was detected by using a horseradish peroxidase (HRP) conjugated secondary antibody and diaminobenzidine (DAB) method that produces a brown color. Cells that not probed with primary antibody were considered as negative controls (A). As it is evident in panel B, NT2 showed an intense nuclear signal for SOX2 expression. (GIF 43.6 kb)

12029_2015_9695_MOESM3_ESM.tif (335 kb)
High Resolution (TIFF 334 kb)

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Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • Mitra Khalili
    • 1
  • Mohammad Vasei
    • 2
  • Davood Khalili
    • 3
  • Kamran Alimoghaddam
    • 4
  • Majid Sadeghizadeh
    • 5
  • Seyed Javad Mowla
    • 5
    Email author
  1. 1.Department of Medical Genetics and Molecular MedicineZanjan University of Medical SciencesZanjanIran
  2. 2.Department of Pathology and Digestive disease research Institute, Shariati HospitalTehran University of Medical SciencesTehranIran
  3. 3.Prevention of Metabolic Disorders Research Center, Research Institute for Endocrine SciencesShahid Beheshti University of Medical SciencesTehranIran
  4. 4.Hematology-Oncology and Stem Cell Research Center, Shariati HospitalTehran University of Medical SciencesTehranIran
  5. 5.Department of Molecular Genetics, Faculty of Biological SciencesTarbiat Modares UniversityTehranIran

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