RETRACTED ARTICLE: MicroRNA-216b is Down-Regulated in Human Gastric Adenocarcinoma and Inhibits Proliferation and Cell Cycle Progression by Targeting Oncogene HDAC8
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Accumulating evidence indicates that micro (mi)RNAs play a critical role in carcinogenesis and cancer progression; however, their role in the tumorigenesis of gastric adenocarcinoma remains unclear so the present study investigated this in gastric cancer (GC) tissues and cell lines.
Human GC specimens (n = 57) and patient-paired non-cancerous specimens were obtained from patients at the First Affiliated Hospital, Henan University of Science and Technology. The AGS and GC9811 gastric cancer cell lines were also used. Expression levels of miR-216b and HDAC8 were examined by quantitative real-time PCR and the expression of HDAC8 was also examined by Western blotting and immunohistochemistry assay. The cell cycle progression was determined by FACS. MiR-216b inhibitor, mimics, and siRNA-HDAC8 transfections were performed to study the loss and gain of function.
We reported a significantly decreased expression of miR-216b in GC clinical specimens compared with paired non-cancerous tissues. We also observed a significant down-regulation of miR-216b expression in GC cell lines AGS and GC9811 (p < 0.0001). The introduction of miR-216b suppressed GC cell proliferation and cell cycle progression by targeting HDAC8, an oncogene shown to promote malignant tumor development with a potential miR-216b binding site in its 3′ untranslated region. HDAC8 expression was shown to be significantly increased in AGS and GC9811 cell lines (p < 0.0001) and GC tissues compared with controls. Moreover, HDAC8 inhibition suppressed cell cycle progression compared with control groups (22 % ± 1.6 % vs 34 % ± 2.1), indicating that HDAC8 may function as an oncogene in the development of GC. Furthermore, HDAC8 expression was negatively correlated (p < 0.0001), while miR-216b expression was positively correlated with the clinical outcome of GC patients (p < 0.0001).
KeywordsGastric Cancer Gastric Cancer Cell Line Human Gastric Cancer HDAC8 Expression GC9811 Cell
Y.W. performed data analyses and wrote the manuscript, initiated the project, designed the experiments, and interpreted the data. P.X. and ZG.S. performed cell culture and cell transfection. SY.S. and XJ.Z. performed qRT-PCR analysis. XS.F. and SG.G performed the cell cycle and proliferation experiments in vitro.
Conflict of Interest
Dr. Ying Wang, Dr. Po Xu, Dr. Jun Yao, Dr. Ruina Yang, Dr. Zhenguo Shi, Dr. Xiaojuan Zhu, Dr. Xiaoshan Feng and Dr. Shegan Gao have no conflicts of interest to disclose.
This study was supported by the National Natural Science Foundation of China (No. 81301763 and No. 81572849) and Henan provincial key scientific and technological projects (No. 142102310473).
Written informed consent was obtained from patients before obtaining tissue samples. The procedures used in this study were approved by the Institutional Review Board of the First Affiliated Hospital, Henan University of Science and Technology and conformed to the Helsinki Declaration and to local legislation.
- 24.Van de Loosdrecht A, Beelen R, Ossenkoppele G, Broekhoven M, Langenhuijsen M (1994) A tetrazolium-based colorimetric MTT assay to quantitate human monocyte mediated cytotoxicity against leukemic cells from cell lines and patients with acute myeloid leukemia. J Immunol Methods 174(1):311–20CrossRefPubMedGoogle Scholar
- 25.Xu X, Zhang Y, Zhang W, Li T, Gao H, Wang Y (2013) MicroRNA-133a functions as a tumor suppressor in gastric cancer. J Biol Regul Homeost Agents 28(4):615–24Google Scholar
- 27.Shen X, Si Y, Yang Z, Wang Q, Yuan J, Zhang X (2015) MicroRNA-542-3p suppresses cell growth of gastric cancer cells via targeting oncogene astrocyte-elevated gene-1. Med Oncol 32(1):1–8Google Scholar
- 29.Zheng X, Dong J, Gong T, Zhang Z, Wang Y, Li Y et al (2015) MicroRNA library-based functional screening identified miR-137 as a suppresser of gastric cancer cell proliferation. J Cancer Res Clin Oncol 141(5):785–95Google Scholar
- 31.Li Z, Yu X, Wang Y, Shen J, Wu WK, Liang J et al (2015) By downregulating TIAM1 expression, microRNA-329 suppresses gastric cancer invasion and growth. Oncotarget 6(19):17559–69Google Scholar
- 32.Han X, Chen Y, Yao N, Liu H, Wang Z (2015) MicroRNA let-7b suppresses human gastric cancer malignancy by targeting ING1. Cancer Gene Ther 22(3):122–9Google Scholar
- 40.Hsieh T-H, Hsu C-Y, Tsai C-F, Long C-Y, Wu C-H, Wu D-C, et al (2015) HDAC inhibitors target HDAC5, upregulate MicroRNA-125a-5p, and induce apoptosis in breast cancer cells. Mol Ther 23(4):656–66Google Scholar
- 43.Chen D-Q, Pan B-Z, Huang J-Y, Zhang K, Cui S-Y, De W et al (2014) HDAC 1/4-mediated silencing of microRNA-200b promotes chemoresistance in human lung adenocarcinoma cells. Oncotarget 5(10):3333–49Google Scholar