Supplementary Figure S2 from miR-204-5p Inhibits Proliferation and Invasion and Enhances Chemotherapeutic Sensitivity of Colorectal Cancer Cells by Downregulating RAB22A
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posted on 2023-03-31, 18:16 authored by Yuan Yin, Binbin Zhang, Weili Wang, Bojian Fei, Chao Quan, Jiwei Zhang, Mingxu Song, Zehua Bian, Qifeng Wang, Shujuan Ni, Yaling Hu, Yong Mao, Leyuan Zhou, Yugang Wang, Jian Yu, Xiang Du, Dong Hua, Zhaohui HuangFigure S2. miR-204-3p showed no significant effect on CRC cell growth. A and B, The expression of miR-204-3p in 6 different CRC cells (A) and tissues (B) was much lower than that of miR-204-5p. C and D, Overexpression of miR-204-3p in LoVo (C) and HCT116 (D) cells did not affect their rates of proliferation. The cell numbers were determined using the Cell Counting Kit-8 assay after transfection. E and F, Inhibition of miR-204-3p in LoVo (E) and HCT116 (F) cells stably expressing miR-204 did not affect their rates of proliferation.
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ARTICLE ABSTRACT
Purpose: miR-204-5p was found to be downregulated in colorectal cancer tissues in our preliminary microarray analyses. However, the function of miR-204-5p in colorectal cancer remains unknown. We therefore investigated the role, mechanism, and clinical significance of miR-204-5p in colorectal cancer development and progression.Experimental Design: We measured the expression of miR-204-5p and determined its correlation with patient prognoses. Ectopic expression in colorectal cancer cells, xenografts, and pulmonary metastasis models was used to evaluate the effects of miR-204-5p on proliferation, migration, and chemotherapy sensitivity. Luciferase assay and Western blotting were performed to validate the potential targets of miR-204-5p after the preliminary screening by a microarray analysis and computer-aided algorithms.Results: miR-204-5p is frequently downregulated in colorectal cancer tissues, and survival analysis showed that the downregulation of miR-204-5p in colorectal cancer was associated with poor prognoses. Ectopic miR-204-5p expression repressed colorectal cancer cell growth both in vitro and in vivo. Moreover, restoring miR-204-5p expression inhibited colorectal cancer migration and invasion and promoted tumor sensitivity to chemotherapy. Mechanistic investigations revealed that RAB22A, a member of the RAS oncogene family, is a direct functional target of miR-204-5p in colorectal cancer. Furthermore, RAB22A protein levels in colorectal cancer tissues were frequently increased and negatively associated with miR-204-5p levels and survival time.Conclusions: Our results demonstrate for the first time that miR-204-5p acts as a tumor suppressor in colorectal cancer through inhibiting RAB22A and reveal RAB22A to be a new oncogene and prognostic factor for colorectal cancer. Clin Cancer Res; 20(23); 6187–99. ©2014 AACR.Usage metrics
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