LncRNA-CR848007.6 regulates the translatome of Cadmium malignant transformed 16HBE cells from a biomechanical perspective
Abstract
Background: Previous studies found that cadmium (Cd) was an environmental toxicant that not only induces toxicological effects but also disrupts cellular biomechanics, affecting cell stiffness, motility, and mechanotransduction pathways. LncRNA-CR848007.6 played an important regulatory role in cadmium toxicology. However, whether its regulatory mechanism involves the biomechanics and changes of the translatome remains to be elucidated. Objective: This study aimed to explore the function and mechanism of LncRNA-CR848007.6 in translation regulation and biomechanical properties during cadmium malignant transformed human bronchial epithelial cells (16HBE cells) through translatome sequencing techniques and bioinformatics methods. Methods: RNA interference technology was applied to silence the expression of LncRNA-CR848007.6 in cadmium malignant transformed 16HBE cells. The libraries of mRNA and RNC were constructed, and Ion Proton™ Sequencer was used for transcriptome and translatome sequencing. Mechanical properties of cells, including stiffness and traction forces, were measured using atomic force microscopy and traction force microscopy. Transcriptome and translatome differential expressions were analyzed using R software. The cell cycle and apoptosis were detected by flow cytometry to verify the functions of LncRNA-CR848007.6 regulating the translatome of cadmium malignant transformed 16HBE cells. Results: Four libraries were obtained after sequencing, including 24062 genes from the transcriptome of the siR mR group cell and NC mR group cell, the translatome of siR RNC group cell and the NC-RNC group cell. It was found that there was little change in the number of transcriptome genes between the siR RNC group cell and NC-RNC group cell, with 19 differentially expressed genes downregulated and 0 differentially expressed genes upregulated. There were 114 genes in the translatome with a ration < −2 and 65 genes in the translatome with a ratio > 2. There was no intersection between the differential TR expression genes and differential mRNA expression genes. The GO analysis results showed significant changes in the translation ratio of cell cycle and mitotic-related pathways, but no enriched KEGG pathway appeared. The cell cycle progression was regulated and cell apoptosis was signficantly inhibited (P < 0.05) after silencing lncRNA-CR848007.6 by siRNA in CdCl2 malignant transformed 16HBE cells. Transcriptome and translatome analyses revealed differential expression of genes involved in cytoskeletal organization and mechanosensitive signaling. Conclusion: LncRNA-CR848007.6 plays a critical role in modulating the biomechanical properties of cadmium-malignant transformed 16HBE cells, influencing cell stiffness and motility through translational regulation. This study provides insights into the biomechanical mechanisms underlying lncRNA-mediated cellular responses to cadmium toxicity.
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