Volume 27, Issue 2 And 3 (3-2023)                   IBJ 2023, 27(2 And 3): 100-107 | Back to browse issues page

XML Print

Download citation:
BibTeX | RIS | EndNote | Medlars | ProCite | Reference Manager | RefWorks
Send citation to:

Akrami H, Shamsdin S A, Nikmanesh Y, Fattahi M R. Effect of Mir-4270 Inhibitor and Mimic on Viability and Stemness in Gastric Cancer Stem-Like Cells Derived from MKN-45 Cell Line. IBJ 2023; 27 (2 and 3) :100-107
URL: http://ibj.pasteur.ac.ir/article-1-3851-en.html
Background: MicroRNAs (miRNAs) are significant regulatory factors in stem cell proliferation, and change in miRNA expression influences the cancer stem cell viability and gene expression. Herein, we evaluated the effect of the hsa-miR-4270 inhibitor and its mimic on the expression of stem cell markers in gastric cancer (GC) stem-like cells.
Methods: GC stem-like cells were isolated from the MKN-45 cell line by a non-adherent surface system. The cells were confirmed by differentiation assays using dexamethasone and insulin as adipogenesis-inducing agents and also staurosporine as a neural-inducing agent. Isolated GC stem-like cells were treated with different concentrations (0, 15, 20, 25, 30, 40, 50, and 60 nM) of hsa-miR-4270 inhibitor and its mimic. The quantity of cell viability was determined by trypan blue method. Transcription of the stem cell marker genes, including CD44, OCT3/4, SOX2, Nanog, and KLF4, was evaluated by real-time RT-PCR.
Results: The results showed that GC stem-like cells were differentiated into both adipose cells using dexamethasone and insulin and neural cells by Staurosporine. Treatment of GC stem-like cells with hsa-miR-4270 inhibitor decreased cell viability and downregulated OCT3/4, CD44, and Nanog to 86%, 79%, and 91% respectively. Also, SOX2 and KLF4 were overexpressed to 8.1- and 1.94-folds, respectively. However, hsa-miR-4270 mimic had opposite effects on the cell viability and gene expression of the stem cell markers.
Conclusion: The effect of hsa-miR-4270 inhibitor and its mimic on the expression of the stem cell markers in GCSCs indicated that hsa-miR-4270 stimulates the stemness property of GCSCs, likely through stimulating the development of gastric stem cells.
Type of Study: Full Length/Original Article | Subject: Cancer Biology

1. Sitarz R, Skierucha M, Mielko J, Offerhaus GJA, Maciejewski R, Polkowski WP. Gastric cancer: epidemiology, prevention, classifcation, and treatment. Cancer management and research 2018; 10: 239-248. [DOI:10.2147/CMAR.S149619]
2. Uchihara T, Ishimoto T, Yonemura A, Baba H. Therapeutic targets against gastric cancer stem cells interacting with tumor microenvironment. Journal of cancer metastasis and treatment 2018; 4(2): 9. [DOI:10.20517/2394-4722.2017.81]
3. Nagini S. Carcinoma of the stomach: A review of epidemiology, pathogenesis, molecular genetics and chemoprevention. World journal of gastrointestinal oncology 2012; 4(7): 156-169. [DOI:10.4251/wjgo.v4.i7.156]
4. Michael Baumann MK, Richard Hill. Exploring the role of cancer stem cells in radioresistance. Nature review 2008; 8: 545-554. [DOI:10.1038/nrc2419]
5. Aponte PM, Caicedo A. Stemness in cancer: Stem cells, cancer stem cells, and their microenvironment. Stem cells international 2017; 2017: 5619472. [DOI:10.1155/2017/5619472]
6. Tannishtha Reya SJM, Michael F. Clarke, Irving L. Weissman. Stem cells, cancer, and cancer stem cells. Nature 2001; 414(1): 105-111. [DOI:10.1038/35102167]
7. Liu J, Ma L, Xu J, Liu C, Zhang J, Liu J, Chen R, Zhou Y. Spheroid body-forming cells in the human gastric cancer cell line MKN-45 possess cancer stem cell properties. Stem cells international 2013; 42(2): 453-459. [DOI:10.3892/ijo.2012.1720]
8. Paolis VD, Lorefice E, Orecchini E, Carissimi C, Laudadio I, Fulci V. Epitranscriptomics: A New layer of microRNA regulation in cancer. Cancers 2021; 13(3372). [DOI:10.3390/cancers13133372]
9. Garg M. MicroRNAs, stem cells and cancer stem cells. World journal of stem cells 2012; 4(7): 62-70. [DOI:10.4252/wjsc.v4.i7.62]
10. Zhu; H-R, Huang; R-Z, Yu; X-N, Shi; X, Bilegsaikhan; E, Guo; H-Y, Song GQ, Weng SQ, Dong L, Janssen HLA, Shen XZ, Zhu JM. Microarray Expression Profling of microRNAs Reveals Potential Biomarkers for Hepatocellular Carcinoma. Tohoku journal of experimental medicine 2018; 245(2): 89-98. [DOI:10.1620/tjem.245.89]
11. Liu J, Ma L, Wang Z, Wang L, Liu C, Chen R, Zhang J. MicroRNA expression profile of gastric cancer stem cells in the MKN-45 cancer cell line. Acta biochimica et biophysica Sinica 2014; 46(2): 92-99. [DOI:10.1093/abbs/gmt135]
12. Golestaneh AF, Atashi A, Langroudi L, Shafiee A, Ghaemi N, Soleimani M. miRNAs expressed differently in cancer stem cells and cancer cells of human gastric cancer cell line MKN-45. Cell biochemistry and function 2012; 30(5): 411-418. [DOI:10.1002/cbf.2815]
13. Wang H, Huang Z, Zhao X, Guo B, Ji Z. miR-4270 regulates cell proliferation and apoptosis in patients with Sertoli cell-only syndrome by targeting GADD45A and inactivating the NOTCH signaling pathway. American journal of translational research 2020; 12(9): 5730-5740.
14. Shen D, Zhao H, Zeng P, Song J, Yang Y, Gu X, Journal of gastric cancer. Circular RNA has-circ-0005556 accelerates gastric cancer progression by sponging miR-4270 to Increase MMP19 expression. Joural of gastric cancer 2020; 20(3): 300-312. [DOI:10.5230/jgc.2020.20.e28]
15. Sun G, Ding X, Bi N, Wang Z, Wu L, Zhou W, Zhao Z, Wang J, Zhang W, Fan J, Zhang W, Dong X, Lv N, Song Y, Zhan Q, Wang L. Molecular predictors of brain metastasisrelated microRNAs in lung adenocarcinoma. PLoS genetics 2019; 15(2): e1007888. [DOI:10.1371/journal.pgen.1007888]
16. Jang BI, Li Y, Graham DY, Cen P. The role of CD44 in the pathogenesis, diagnosis, and therapy of gastric cancer. Gut and liver 2011; 5(4): 397. [DOI:10.5009/gnl.2011.5.4.397]
17. van Schaijik B, Davis PF, Wickremesekera AC, Tan ST, Itinteang T. Subcellular localisation of the stem cell markers OCT4, SOX2, NANOG, KLF4 and c-MYC in cancer: a review. Journal of clinical pathology 2018; 71(1): 88-91. [DOI:10.1136/jclinpath-2017-204815]
18. Fatemeh Mahmoodi, Akrami H. PlGF Knockdown decreases tumorigenicity and stemness properties of spheroid body cells derived from gastric cancer cells. Journal of cellular biochemistry 2017; 118(4): 851-859. [DOI:10.1002/jcb.25762]
19. Zengfu Xue HY, Juntang Li, Shuli Liang, Xiqiang Cai, Xiong Chen, Qiong Wu, Liucun Gao, Kaichun Wu, Yongzhan Nie, Daiming Fan. Identification of cancer stem cells in vincristine preconditioned SGC7901 gastric cancer cell line. Journal of cellular biochemistry 2012; 113: 302-312. [DOI:10.1002/jcb.23356]
20. Xiaomeng Xu XZ, Sheng Wang ,Hui Qian ,Wei Zhu, Huiling Cao ,Mei Wang ,Yuan Chen ,Wenrong Xu. Isolation and comparison of mesenchymal stem-like cells from human gastric cancer and adjacent non-cancerous tissues. Journal of cancer research and clinical oncology 2011; 137: 495-504. [DOI:10.1007/s00432-010-0908-6]
21. Chen C, Ridzon DA, Broomer AJ, Zhou Z, Lee DH, Nguyen JT, Barbisin M, Lan Xu N, Mahuvakar VR, Andersen MR, Lao KQ, Livak KJ, Guegler KJ. Real-time quantification of microRNAs by stem-loop RT-PCR. Nucleic acids research 2005; 33(20): e179. [DOI:10.1093/nar/gni178]
22. Akrami H, Moradi B, Borzabadi Farahani D, Mehdizadeh K. Ibuprofen reduces cell proliferation through inhibiting Wnt/β catenin signaling pathway in gastric cancer stem cells. Cell biology international 2018; 42(8): 949-958. [DOI:10.1002/cbin.10959]
23. Livak KJ, Schmittgen TD. Analysis of relative gene expression data using realtime quantitative PCR and the 2-DDCT method. Methods 2001; 25: 402-408. [DOI:10.1006/meth.2001.1262]
24. Peitzsch C, Tyutyunnykova A, Pantel K, Dubrovska A. Cancer stem cells: The root of tumor recurrence and metastases. Seminars in cancer biology 2017; 44: 10-24. [DOI:10.1016/j.semcancer.2017.02.011]
25. Xia P, Xu XY. Epithelial-mesenchymal transition and gastric cancer stem cell. Tumour biology 2017; 39(5): 1010428317698373. [DOI:10.1177/1010428317698373]
26. Fu L, Bu L, Yasuda T, Koiwa M, Akiyama T, Uchihara T, Baba H, Ishimoto T. Gastric cancer stem cells: current insights into the immune microenvironment and therapeutic targets. Biomedicines 2020; 8(1): 1-10. [DOI:10.3390/biomedicines8010007]
27. Huiwen Yan , Bu P. Non-coding RNAs in cancer stem cells. Cancer letters 2018; 421: 121-126. [DOI:10.1016/j.canlet.2018.01.027]
28. Peng W Z, Ma R, Wang F, Yu J, Liu Z B. Role of miR-191/425 cluster in tumorigenesis and diagnosis of Gastric Cancer. International journal of molecular sciences 2014; 15(3): 4031-448. [DOI:10.3390/ijms15034031]
29. Huang Q, Gumireddy K, Schrier M, Sage Cl, Nagel R, Nair S, Egan DA, Li A, Huang G, Klein Szanto AJ, Gimotty PA, Katsaros D, Coukos G, Zhang L, PuréE, Agami R. The microRNAs miR-373 and miR-520c promote tumour invasion and metastasis. Nature cell biology 2008; 10(2): 202-210. [DOI:10.1038/ncb1681]
30. Mahmoodi F, Akrami H. PlGF Knockdown decreases tumorigenicity and stemness properties of spheroid body cells derived from gastric cancer cells. Journal of cellular biochemistry 2017; 118(4): 851-859. [DOI:10.1002/jcb.25762]
31. Zhang Z, Li Z, Li Y, Zang A. MicroRNA and signaling pathways in gastric cancer. Cancer gene therapy 2014; 21(8): 305-316. [DOI:10.1038/cgt.2014.37]
32. Tay Y, Zhang J, Thomson AM, Lim B, Rigoutsos I. MicroRNAs to Nanog, Oct4 and Sox2 coding regions modulate embryonic stem cell differentiation. Nature 2008; 455(7216): 1124-1128. [DOI:10.1038/nature07299]
33. Otsubo T, Akiyama Y, Hashimoto Y, Shimada S, Goto K, Yuasa Y. MicroRNA-126 inhibits SOX2 expression and contributes to gastric carcinogenesis. PloS one 2011; 6(1): e16617. [DOI:10.1371/journal.pone.0016617]

Add your comments about this article : Your username or Email:

Rights and permissions
Creative Commons License This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.

© 2024 CC BY-NC 4.0 | Iranian Biomedical Journal

Designed & Developed by : Yektaweb