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Abstract：
OBJECTIVE: Ribonuclease P RNA component H1 (RPPH1) is a long non-coding RNA (lncRNA) associated with cancer progression. Higher RPPH1 expression in breast and cervical cancer samples than that in normal tissues were observed through the lncRNASNP2 database; therefore, silencing RPPH1 expression might be a potential strategy for cancer treatments, even though RPPH1 is also an RNA subunit of ribonuclease P involved in processing transfer RNA (tRNA) precursors and the effect of RPPH1 knockdown is not yet fully understood.
METHODS: Differentially expressed genes (DEGs) were identified through RNA sequencing in each shRNA-transfected RPPH1 knockdown MDA-MB-231, RPPH1 knockdown HeLa cell, and respective control cells, then the gene ontology enrichment analysis was performed by IPA and MetaCore database according to these DEGs, with further in vitro experiments validating the effect of RPPH1 silencing in MDA-MB-231 and HeLa cells.
RESULTS: Hundreds of down-regulated DEGs were identified in RPPH1 knockdown MDA-MB-231 and HeLa cells while bioinformatics analysis revealed that these genes were involved in pathways related to immune response and cancerogenesis. Compared to mock- and vector-transfected cells, the production of mature tRNAs, cell proliferation and migration capacity were inhibited in RPPH1-silenced HeLa and MDA-MB-231 cells. Additionally, RPPH1 knockdown promoted G1 cell cycle arrest mainly through the down-regulation of cyclin D1, although glycolytic pathways were only affected in RPPH1 knockdown HeLa cells but not MDA-MB-231 cells.
CONCLUSIONS: This study demonstrated that knockdown RPPH1 affected tRNA production, cell proliferation and metabolism. Our findings might provide insight into the role of RPPH1 in tumor development.
METHODS: Differentially expressed genes (DEGs) were identified through RNA sequencing in each shRNA-transfected RPPH1 knockdown MDA-MB-231, RPPH1 knockdown HeLa cell, and respective control cells, then the gene ontology enrichment analysis was performed by IPA and MetaCore database according to these DEGs, with further in vitro experiments validating the effect of RPPH1 silencing in MDA-MB-231 and HeLa cells.
RESULTS: Hundreds of down-regulated DEGs were identified in RPPH1 knockdown MDA-MB-231 and HeLa cells while bioinformatics analysis revealed that these genes were involved in pathways related to immune response and cancerogenesis. Compared to mock- and vector-transfected cells, the production of mature tRNAs, cell proliferation and migration capacity were inhibited in RPPH1-silenced HeLa and MDA-MB-231 cells. Additionally, RPPH1 knockdown promoted G1 cell cycle arrest mainly through the down-regulation of cyclin D1, although glycolytic pathways were only affected in RPPH1 knockdown HeLa cells but not MDA-MB-231 cells.
CONCLUSIONS: This study demonstrated that knockdown RPPH1 affected tRNA production, cell proliferation and metabolism. Our findings might provide insight into the role of RPPH1 in tumor development.
摘要：
目的：核糖核酸酶PRNA组分H1（RPPH1）是一种与癌症进展相关的长链非编码RNA（lncRNA）。通过lncRNASNP2数据库观察到乳腺癌和宫颈癌样本中的RPPH1表达高于正常组织;因此,沉默RPPH1表达可能是癌症治疗的潜在策略，尽管RPPH1也是核糖核酸酶P的RNA亚基，参与加工转移RNA（tRNA）前体，但RPPH1敲低的作用尚未完全了解。
方法：通过RNA测序在每个shRNA转染的RPPH1敲低MDA-MB-231，RPPH1敲低HeLa细胞中鉴定差异表达基因（DEGs），和各自的对照细胞，然后根据这些DEG通过IPA和MetaCore数据库进行基因本体富集分析，进一步的体外实验验证了RPPH1沉默在MDA-MB-231和HeLa细胞中的作用。
结果：在RPPH1敲低MDA-MB-231和HeLa细胞中鉴定出数百个下调的DEGs，而生物信息学分析显示这些基因参与与免疫应答和癌症发生相关的通路。与模拟和载体转染的细胞相比，成熟tRNA的产生，在RPPH1沉默的HeLa和MDA-MB-231细胞中，细胞增殖和迁移能力受到抑制。此外,RPPH1敲低主要通过下调细胞周期蛋白D1促进G1细胞周期阻滞，尽管糖酵解途径仅在RPPH1敲低HeLa细胞中受影响，而在MDA-MB-231细胞中不受影响。
结论：这项研究表明敲低RPPH1影响tRNA的产生，细胞增殖和代谢。我们的发现可能为了解RPPH1在肿瘤发展中的作用提供了见解。
方法：通过RNA测序在每个shRNA转染的RPPH1敲低MDA-MB-231，RPPH1敲低HeLa细胞中鉴定差异表达基因（DEGs），和各自的对照细胞，然后根据这些DEG通过IPA和MetaCore数据库进行基因本体富集分析，进一步的体外实验验证了RPPH1沉默在MDA-MB-231和HeLa细胞中的作用。
结果：在RPPH1敲低MDA-MB-231和HeLa细胞中鉴定出数百个下调的DEGs，而生物信息学分析显示这些基因参与与免疫应答和癌症发生相关的通路。与模拟和载体转染的细胞相比，成熟tRNA的产生，在RPPH1沉默的HeLa和MDA-MB-231细胞中，细胞增殖和迁移能力受到抑制。此外,RPPH1敲低主要通过下调细胞周期蛋白D1促进G1细胞周期阻滞，尽管糖酵解途径仅在RPPH1敲低HeLa细胞中受影响，而在MDA-MB-231细胞中不受影响。
结论：这项研究表明敲低RPPH1影响tRNA的产生，细胞增殖和代谢。我们的发现可能为了解RPPH1在肿瘤发展中的作用提供了见解。
