PRKCSH Alternative Splicing Involves in Silica-Induced Expression of Epithelial–Mesenchymal Transition Markers and Cell Proliferation

Abstract

Background: Mounting evidence suggests that alternative splicing is one of the ways for cells to adapt to environmental stress insults. The aim of this study was firstly to examine the effect of silica on the alternative splicing of lung fibrosis–associated genes. Methods: Microarray analysis was used to construct the alternative splicing profile. Functional experiments were conducted using Cell Counting Kit-8, cell cycle, apoptosis, and epithelial–mesenchymal transition (EMT) analyses. Alternative splicing variants were verified by quantitative real-time polymerase chain reaction (qRT-PCR) polymerase chain reaction method. Results: A total of 1850 genes that have alternative splices in response to silica insult were identified. PCDHB11, MALAT1, MT2A, RP11-126D17.1, and RP11-415I12.2 are the top 5 upregulated genes with occurrence of alternative splice, whereas NDE1, RNPEPL1, TREML2, CSF2RB, and PRKCSH are the top 5 downregulated genes with occurrence of alternative splice. Bioinformatic analysis showed these genes with the occurrence of alternative splice mainly are associated with EMT pathway, N-Glycan biosynthesis, and leukocyte transendothelial migration. Further study indicated that PRKCSH-2 knockdown promotes A549 cell proliferation potential by partially promoting EMT signals. Conclusions: Significant changes in alternative splicing of silicosis-associated genes occur in patients with silicosis in silica conditions. Our study provides basic founding for further investigation into the detail molecular mechanisms underlying silica-induced silicosis.

Transcriptomic and metabolomic profiling reveal the p53-dependent benzeneacetic acid attenuation of silica‐induced epithelial–mesenchymal transition in human bronchial epithelial cells

“…1 a). We harvested cells for transcriptomic detection based on our findings from our pilot study that suggested that cell damage at this timepoint could significantly induce EMT [ 29 – 31 ]. The details of the changes in mRNA expression are listed in Table 1 .…”

“…WB detection was conducted as previously reported [ 22 , 28 , 29 ]. In this study, antibodies against p53, E-cadherin, N-cadherin, and Vimentin were purchased from Cell Signaling Technology, Inc.(Danvers, MA, USA) and Abcam (Cambridge, UK).…”

Transcriptomic and Metabolomic Profiling Reveal the p53-Dependent Benzeneacetic Acid Attenuation of Silica-Induced Epithelial–Mesenchymal Transition in Human Bronchial Epithelial Cells

“…WB detection was conducted as previously reported [22,28,29]. In this study, antibodies against p53, E-cadherin, N-cadherin, and vimentin were purchased from Cell Signaling Technology, Inc.(Danvers, MA, USA) and Abcam (Cambridge, UK).Images were captured and assessed using the ChemiDocXRS + system (Bio-Rad Laboratories).…”

“…1A). We harvested cells for transcriptomic detection based on our ndings from our pilot study that suggested that cell damage at this timepoint could signi cantly induce EMT [29][30][31]. The details of the changes in mRNA expression are listed in Table 1.…”

Potential role of PRKCSH in lung cancer: bioinformatics analysis and a case study of Nano ZnO

“…13 We previously reported that PRKCSH expression increased in response to silica-induced epithelial–mesenchymal transition and that the knockdown (KD) of a PRKCSH-2 splice variant stimulated A549 cell proliferation by promoting epithelial–mesenchymal transition. 14 It remains unclear whether PRKCSH interacts with lung cancer immune infiltration-related signalling pathways and/or whether this affects lung cancer proliferation or prognosis.…”