American Association for Cancer Research
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supplemental Figure legends from Long Noncoding RNA GCASPC, a Target of miR-17-3p, Negatively Regulates Pyruvate Carboxylase–Dependent Cell Proliferation in Gallbladder Cancer

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posted on 2023-03-31, 00:29 authored by Ming-zhe Ma, Yan Zhang, Ming-zhe Weng, Shou-hua Wang, Ye Hu, Zhao-yuan Hou, Yi-yu Qin, Wei Gong, Yong-Jie Zhang, Xiang Kong, Jian-dong Wang, Zhi-wei Quan

supplemental Figure 1-12. Expression levels of 10 lncRNAs by qRT-PCR in 15 GBC tissues and paired non-tumor tissues (Figure 1). Expression levels of 8 mRNAs by qRT-PCR in 15 GBC tissues and paired non-tumor tissues (Figure 2). Molecular characteristics of GCASPC (Figure 3). Nucleotide sequence of the full-length human GCASPC gene (Figure 4). GCASPC has no coding capability (Figure 5). The expression level of GCASPC in gallbladder cancer cells (Figure 6). Manipulation of GCASPC expression level (Figure 7). Anti-proliferative activity of GCASPC (Figure 8). GCASPC acts in trans (Figure 9). The effect of GCASPC on the protein, activity and mRNA levels of PC (Figure 10). GCASPC destabilizes PC protein (Figure 11). GCASPC directly binds to miR-17-3p (Figure 12).

Funding

National Natural Science Foundation of China

Key University Science Research Project of Anhui Province

History

ARTICLE ABSTRACT

Long noncoding RNAs (lncRNA) are being implicated in the development of many cancers. Here, we report the discovery of a critical role for the lncRNA GCASPC in determining the progression of gallbladder cancer. Differentially expressed lncRNAs and mRNAs between gallbladder cancer specimens and paired adjacent nontumor tissues from five patients were identified and validated by an expression microarray analysis. Quantitative real-time PCR was used to measure GCASPC levels in tissues from 42 gallbladder cancer patients, and levels of GCASPC were confirmed further in a separate cohort of 89 gallbladder cancer patients. GCASPC was overexpressed or silenced in several gallbladder cancer cell lines where molecular and biological analyses were performed. GCASPC levels were significantly lower in gallbladder cancer than adjacent nontumor tissues and were associated with tumor size, American Joint Committee on Cancer tumor stage, and patient outcomes. GCASPC overexpression suppressed cell proliferation in vitro and in vivo, whereas GCASPC silencing had opposite effects. By RNA pull-down and mass spectrometry, we identified pyruvate carboxylase as an RNA-binding protein that associated with GCASPC. Because GCASPC is a target of miR-17-3p, we confirmed that both miR-17-3p and GCASPC downregulated pyruvate carboxylase level and activity by limiting protein stability. Taken together, our results defined a novel mechanism of lncRNA-regulated cell proliferation in gallbladder cancer, illuminating a new basis for understanding its pathogenicity. Cancer Res; 76(18); 5361–71. ©2016 AACR.