Abstract
Investigation on a competitive endogenous RNA (ceRNA) network attracted lots of attention due its function in cancer regulation. Here, we probed into the possible molecular mechanism of circSSPO/microRNA-6820-5p (miR-6820-5p)/kallikrein-related peptidase 8 (KLK8)/PKD1 network in the esophageal squamous cell carcinoma (ESCC). Following whole-transcriptome sequencing and differential analysis in collected ESCC tissue samples, circRNA-miRNA-mRNA regulatory network affecting ESCC was investigated. After interaction measurement among circSSPO/miR-6820-5p/KLK8/PKD1, their regulatory roles in ESCC cell functions in vitro and xenograft tumor growth and lung metastasis in vivo were analyzed. The bioinformatics prediction and sequencing results screened that circSSPO, miR-6820-5p, KLK8, and PKD1 were associated with ESCC development. In ESCC, miR-6820-5p was expressed at very low levels, while circSSPO, KLK8, and PKD1 were highly expressed. In vitro cell experiments further proved that circSSPO competitively inhibited miR-6820-5p to induce ESCC cell malignant properties. Moreover, knockdown of KLK8 or PKD1 inhibited ESCC cell malignant properties. circSSPO also promoted the tumorigenic and metastasis of ESCC through the upregulation of KLK8 and PKD1 expression in vivo. We found that circSSPO was an oncogenic circRNA that was significantly abundant in ESCC tissues and circSSPO exhibited an oncogenic activity in ESCC by elevating expression of KLK8 and PKD1 through suppressing miR-6820-5p expression.
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Funding
1. Primary Health Care Foundation of China-Clinical Application Research and Medical Training Fund Project Key Project (No: 2021120002). 2. 2022 Guangzhou Basic Research Plan, basic and applied basic research projects jointly funded by the city and schools (college) (No: 202201020078). 3. Guangdong Science and Technology special fund in 2020 (Shan Fu Ke letter (2020) 53; Project ID: 200113165875501 and 200115105879476). 4. 2022 Guangdong Science and Technology Innovation Strategic Special project (“ Major projects + Task List “) [Shan Fu Ke letter (2022) 124, Project ID: STKJ202209072]. 5. Research-oriented Hospital Program of Guangzhou [RHPG05].
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Conceived and designed the experiments: Qianhua Luo, Junzheng Li, Haixiong Miao, Siman Su, Chengcheng Xu. Performed experimental: Yun Chen, Chengkuan Zhao, Jianxiang Huang. Performed experimental validation: Kai Ling. Analyzed the data and conceived figures and tables: Yun Chen, Chengkuan Zhao, Jianxiang Huang. Wrote the paper: Qianhua Luo, Junzheng Li, Haixiong Miao, Siman Su, Chengcheng Xu. Contributed to the writing of the manuscript: Chaoxian Lin, Hongfei Yan, Shuyao Zhang. All authors revised and approved the final version of the manuscript.
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This study complied with the ethical guidelines of the Declaration of Helsinki and was approved by the Clinical Ethics Committee of Guangzhou Red Cross Hospital of Jinan University (No. 2021-083-02). All enrolled patients provided written informed consent. All animal experiments were approved by the ethics committee of Guangzhou Red Cross Hospital of Jinan University.
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Simple summary: This study was performed to analyze the role of circSSPO in esophageal squamous cell (ESCC).
Supplementary Information
ESM 1
Supplementary Figure 1. Identification of circSSPO. A, The chromosomal positioning and Sanger sequencing results of circSSPO. The black arrow marks the reverse splice point of the circSSPO. B, Agarose gel electrophoresis results of the cDNA and gDNA amplified with divergent and convergent primers in TE-1 and EC9706 cells. C, After the treatment of TE-1 and EC9706 cells with RNase R, levels of linear SSPO mRNA and circSSPO were determined by RT-qPCR. * p < 0.05, compared with the Mock group. D, After the treatment of TE-1 and EC9706 cells with actinomycin D, the abundance of linear SSPO mRNA and circSSPO was measured by RT-qPCR. * p < 0.05, compared with 0 h. Cell experiments were repeated three times independently. (JPG 766 kb)
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Luo, Q., Li, J., Miao, H. et al. circSSPO boosts growth of esophageal squamous cell carcinoma through upregulation of micrRNA-6820-5p-mediated KLK8 and PKD1 expression. Cell Biol Toxicol 39, 3219–3234 (2023). https://doi.org/10.1007/s10565-023-09828-3
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DOI: https://doi.org/10.1007/s10565-023-09828-3