American Association for Cancer Research
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Supplementary Figure S3 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 Huang

Figure S3. Ectopic miR-204-5p expression increases the sensitivity of CRC cells to 5-FU and DDP. A, and B, miR-204 overexpression enhanced 5-FU and DDP-induced apoptosis in LoVo (A) and HCT116 (B) cells. Annexin V/PI staining and FACs analysis were performed to detect apoptosis of LoVo and HCT116 cells after administration of 5 μg/mL 5-FU and DDP. The apoptosis levels were determined by the Annexin V positive ratio.



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.

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