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.

中文翻译：

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PRKCSH 可变剪接涉及二氧化硅诱导的上皮-间质转化标志物的表达和细胞增殖。

背景：

越来越多的证据表明，选择性剪接是细胞适应环境压力损伤的方式之一。本研究的目的首先是检查二氧化硅对肺纤维化相关基因可变剪接的影响。

方法：

微阵列分析用于构建可变剪接谱。使用 Cell Counting Kit-8、细胞周期、细胞凋亡和上皮-间质转化 (EMT) 分析进行功能实验。通过定量实时聚合酶链反应 (qRT-PCR) 聚合酶链反应方法验证可变剪接变体。

结果：

共鉴定了 1850 个响应二氧化硅损伤而具有选择性剪接的基因。PCDHB11、MALAT1、MT2A、RP11-126D17.1 和 RP11-415I12.2 是发生选择性剪接的前 5 个上调基因，而 NDE1、RNPEPL1、TREML2、CSF2RB 和 PRKCSH 是发生选择性剪接的前 5 个下调基因替代拼接。生物信息学分析表明，这些可变剪接发生的基因主要与EMT途径、N-聚糖生物合成和白细胞跨内皮迁移有关。进一步的研究表明，PRKCSH-2 敲低通过部分促进 EMT 信号来促进 A549 细胞的增殖潜力。

结论：

矽肺相关基因选择性剪接的显着变化发生在二氧化硅条件下的矽肺患者身上。我们的研究为进一步研究二氧化硅诱导矽肺的详细分子机制提供了基础。