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©The Author(s) 2015. Published by Baishideng Publishing Group Inc. All rights reserved.
Primary analysis and screening of microRNAs in gastric cancer side population cells
Hai-Hong Zhang, Guo-Li Gu, Xiang-Yang Zhang, Feng-Zhi Li, Li Ding, Qin Fan, Ran Wu, Wei Shi, Xin-Yan Wang, Lin Chen, Xue-Ming Wei, Xiao-Ying Yuan
Hai-Hong Zhang, Guo-Li Gu, Xue-Ming Wei, Department of General Surgery, Air Force General Hospital of Chinese PLA, Beijing 100142, China
Xiang-Yang Zhang, Feng-Zhi Li, Li Ding, Qin Fan, Wei Shi, Xin-Yan Wang, Xiao-Ying Yuan, Department of Intelligence, Air Force General Hospital of Chinese PLA, Beijing 100142, China
Lin Chen, Department of General Surgery, Chinese PLA General Hospital, Beijing 100853, China
Ran Wu, Department of Internal Medicine, Northern affiliated Hospital of China North Industries Group Corporation, Beijing 100089, China
Author contributions: Zhang HH, Wei XM, Gu GL and Chen L designed the research; Zhang HH, Zhang XY, Ding L, Li FZ and Fan Q performed the research; Zhang XY and Wang XY provided the analytic tools; Zhang HH, Zhang XY, Shi W and Fan Q analyzed the data; Zhang HH, Wu R, Chen L, Yuan XY and Li FZ wrote the manuscript; Wei XW and Yuan XY contributed equally to this work; Zhang HH and Gu GL contributed equally to this work.
Supported by National Natural Science Fund, No. 81000706/H1108; National Key Technology Research and Development Program, No. 2012BA141B01; and the Air Force General Hospital of Chinese PLA Innovation Fund, No. KZ2013035.
Ethics approval: The study was reviewed and approved by the Medical Ethics Committee of PLA Air Force General Hospital.
Conflict-of-interest: There are no conflicts of interest to declare.
Data sharing: No additional data are available.
Open-Access: This article is an open-access article which was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution Non Commercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See: http://creativecommons.org/licenses/by-nc/4.0/
Correspondence to: Xue-Ming Wei, Professor, Department of General Surgery, Air Force General Hospital of Chinese PLA, No. 30 Fucheng Road, Haidian District, Beijing 100142, China.
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Received: October 29, 2014
Peer-review started: October 29, 2014
First decision: November 14, 2014
Revised: December 22, 2014
Accepted: January 16, 2015
Article in press: January 16, 2015
Published online: March 28, 2015
Processing time: 151 Days and 21.5 Hours
AIM: To explore the microRNA (miRNA) profiles and to determine the key miRNAs within the side population (SP) cells of the gastric cancer cell line MKN-45.
METHODS: We used fluorescence-activated cell sorting and Hoechst 33342 labeling to obtain SP cells from the human gastric carcinoma cell line MKN-45. The miRNA expression profiles of the SP and major population (MP) cells were examined using a miRNA gene chip, and key miRNAs were obtained according to aberrant expression and the miRNAs’ possible targets as predicted by bioinformatics.
RESULTS: Using a significance criterion of a 1.5-fold or greater difference in expression level, we observed an increase in the expression of 34 miRNAs and a decrease in the expression of 34 miRNAs when comparing SP to MP cells. Using quantitative real-time reverse transcription-polymerase chain reaction to test for differentially expressed miRNAs combined with bioinformatics results, we found that the downregulated miRNAs, such as hsa-miR-3175 and hsa-miR-203, and the upregulated miRNAs, including hsa-miR-130a, hsa-miR-324-5p, hsa-miR-34a, and hsa-miR-25-star, may be important in maintaining and regulating the characteristics of SP cells.
CONCLUSION: There are key miRNAs expressed within the SP cells of the gastric cancer cell line MKN-45, and include hsa-miR-3175, hsa-miR-203, hsa-miR-130a, hsa-miR-324-5p, hsa-miR-34a, and hsa-miR-25-star.
Core tip: MicroRNAs (miRNAs) act as one mechanism by which coding genes are regulated, and they are involved in many biological and pathological processes including carcinogenesis. However, identifying specific miRNAs is difficult, because many miRNAs possess more than one target and many target genes are regulated by more than one miRNA. We used fluorescence-activated cell sorting and Hoechst 33342 labeling to obtain side population (SP) cells from the human gastric carcinoma cell line MKN-45. The miRNA expression profiles of SP and major population cells were examined by miRNA gene chip analysis, and key miRNAs were identified according to aberrant expression and their possible targets as predicted by bioinformatics.
