Liming Jin1,2,3, Zhaoxia Zhang1,2,3, Zhang Wang2,3, Xiaojun Tan2,3, Zhaoying Wang2,3, Lianju Shen2,3, Chunlan Long2,3, Guanghui Wei1,2,3, Dawei He4,5,6. 1. Department of Urology, Children's Hospital of Chongqing Medical University, Chongqing, 400014, China. 2. Chongqing Key Laboratory of Children Urogenital Development and Tissue Engineering, Chongqing, 400014, China. 3. China International Science and Technology Cooperation Base of Child Development and Critical, Ministry of Education Key Laboratory of Child Development and Disorders; Chongqing Key Laboratory of Pediatrics, National Clinical Research Center for Child Health and Disorders, ChongqingChongqing, 400014, China. 4. Department of Urology, Children's Hospital of Chongqing Medical University, Chongqing, 400014, China. hedawei@hospital.cqmu.edu.cn. 5. Chongqing Key Laboratory of Children Urogenital Development and Tissue Engineering, Chongqing, 400014, China. hedawei@hospital.cqmu.edu.cn. 6. China International Science and Technology Cooperation Base of Child Development and Critical, Ministry of Education Key Laboratory of Child Development and Disorders; Chongqing Key Laboratory of Pediatrics, National Clinical Research Center for Child Health and Disorders, ChongqingChongqing, 400014, China. hedawei@hospital.cqmu.edu.cn.
Abstract
BACKGROUND: CSCs play an important role in tumor development. Some studies have demonstrated that piRNAs participate in the progression of various cancers. However, the detailed function of piRNAs in CSCs requires further investigation. This study aimed to investigate the significance of novel piRNA MW557525, one of the top five up-regulated piRNAs screened by gene chip and it has been verified by RT-q-PCR that it is indeed the most obvious up-regulated expression in Piwil2-iCSCs. METHODS AND RESULTS: Differentially expressed piRNAs in Piwil2-iCSCs were screened by gene chip. Target genes were predicted by the miRanda algorithm and subjected to GO and KEGG analysis. One of the differential piRNAs, novel piRNA MW557525, was transfected and its target gene NOP56 was silenced in Piwil2-iCSCs, respectively. RT-qPCR, western blot (WB) and dual luciferase reporter assay were used to investigate the interaction of piRNA MW557525 and NOP56. We identified the effect of piRNA MW557525 and NOP56 knockdown on cell proliferation, migration, invasion, and apoptosis via CCK-8, transwell assay, and flow cytometry. The expressions of CD24, CD133, KLF4, and SOX2 were detected via WB. The results showed that piRNA MW557525 was negatively correlated with NOP56, and it promoted the proliferation, migration, invasion, and inhibited apoptosis in Piwil2-iCSCs, and it also promoted the expressions of CD24, CD133, KLF4, and SOX2, while NOP56 showed the opposite effect. CONCLUSIONS: These findings suggested that novel piRNA MW557525 might be a novel therapeutic target in Piwil2-iCSCs.
BACKGROUND: CSCs play an important role in tumor development. Some studies have demonstrated that piRNAs participate in the progression of various cancers. However, the detailed function of piRNAs in CSCs requires further investigation. This study aimed to investigate the significance of novel piRNA MW557525, one of the top five up-regulated piRNAs screened by gene chip and it has been verified by RT-q-PCR that it is indeed the most obvious up-regulated expression in Piwil2-iCSCs. METHODS AND RESULTS: Differentially expressed piRNAs in Piwil2-iCSCs were screened by gene chip. Target genes were predicted by the miRanda algorithm and subjected to GO and KEGG analysis. One of the differential piRNAs, novel piRNA MW557525, was transfected and its target gene NOP56 was silenced in Piwil2-iCSCs, respectively. RT-qPCR, western blot (WB) and dual luciferase reporter assay were used to investigate the interaction of piRNA MW557525 and NOP56. We identified the effect of piRNA MW557525 and NOP56 knockdown on cell proliferation, migration, invasion, and apoptosis via CCK-8, transwell assay, and flow cytometry. The expressions of CD24, CD133, KLF4, and SOX2 were detected via WB. The results showed that piRNA MW557525 was negatively correlated with NOP56, and it promoted the proliferation, migration, invasion, and inhibited apoptosis in Piwil2-iCSCs, and it also promoted the expressions of CD24, CD133, KLF4, and SOX2, while NOP56 showed the opposite effect. CONCLUSIONS: These findings suggested that novel piRNA MW557525 might be a novel therapeutic target in Piwil2-iCSCs.
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