Abstract

BackgroundOur previous study described the crucial role of Rho-associated coiled-coil containing-kinases (ROCK) in hepatocellular carcinoma (HCC). However, the potential significance of long noncoding RNA downstream of ROCK is largely unknown. Here, a comprehensive comparative bioinformatics analysis of a microarray of an MHCC-97H cell line overexpressing ROCK1 or ROCK2 was performed.ResultsNumerous lncRNAs and mRNAs were deregulated by Rho-associated coiled-coil containing kinases 1 and 2. These results were consistent with the qRT-PCR results. Compared with MHCC-97H-Con, which was transfected with a null vector, the GO analysis revealed differentially expressed mRNAs (DEmRNAs) in MHCC-97H-ROCK1 (ROCK1 was overexpressed) enriched in apoptotic cell clearance, the cyclooxygenase pathway and bone trabecula morphogenesis; the DEmRNAs in MHCC-97H-ROCK2 (ROCK2 was overexpressed) were enriched in VEGF production, chemokine-associated signaling pathways, acute inflammatory response and vasoconstriction. Compared with MHCC-97H-ROCK2, the DEmRNAs in MHCC-97H-ROCK1 were involved in the JAK-STAT cascade, the Akt signaling pathway and the activity of several different peptidases. The pathway analysis of ROCK1 and ROCK2 revealed an overlap in the VEGF signaling pathway, ECM-receptor interaction, and adhesion and differences in the PPAR signaling pathway and mismatch repair. The predicted targets of the differentially expressed lncRNA (DElncRNAs) were enriched in the p53 signaling pathway, Jak-STAT signaling pathway, etc.Several hub DElncRNAs were identified.ConclusionsROCK1 and 2 modulate the expression of numerous mRNAs and lncRNAs and may participate in several signaling pathways in HCC. Several hub molecules were identified in the lncRNA-mRNA networks. Our results provide baseline data for ROCK1 and 2 regulation in HCC that might have implications for further research.

Highlights

  • Our previous study described the crucial role of Rho-associated coiled-coil containing-kinases (ROCK) in hepatocellular carcinoma (HCC)

  • ROCK isoforms were overexpressed in MHCC-97H ROCK1 and ROCK2 stably expressing cells were established

  • Using a method of Limma and level of significance of p < 0.05 and a fold change (FC) > 2, 840 differentially expressed messenger RNA (mRNA) (DEmRNAs) were identified in the MHCC-97H-ROCK1 cells, including 183 (21.79%) downregulated and 657 (78.21%) upregulated DEmRNAs, and 1022 DElncRNAs were identified in the ROCK1 upregulated samples, including 303 (29.65%) upregulated and 719 (70.35%) downregulated DElncRNAs

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Summary

Introduction

Our previous study described the crucial role of Rho-associated coiled-coil containing-kinases (ROCK) in hepatocellular carcinoma (HCC). Several studies have reported that long non coding RNAs (lncRNAs) may regulate the cytoskeleton by ROCK signaling during tumor migration and metastasis, indicating that ROCK are associated with lncRNA during key steps in carcinoma [11,12,13]. Similar to its broad effects on global miRNA transcriptional expression, these studies have suggested that the activity of ROCK1 and 2 may affect the level of several coding and long noncoding RNAs. To further understand the functional significance of mRNAs and lncRNAs in ROCK-mediated pathological processes in HCC, RNA sequencing technology was utilized to profile the mRNA and lncRNA expressional signatures in three pairs of ROCK1 overexpression, ROCK2 overexpression and normal control MHCC-97H cell lines, which have a high metastatic ability [14]. Our study focused on differentially expressed mRNAs (DEmRNAs) and lncRNAs (DElncRNAs) located downstream of Rho/ ROCK pathways and formulated network charts to identify crucial nodal molecules

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