The aberrant upregulation of exon 10-inclusive SREK1 through SRSF10 acts as an oncogenic driver in human hepatocellular carcinoma. The aberrant upregulation of exon 10-inclusive SREK1 through SRSF10 acts as an oncogenic driver in human hepatocellular carcinoma

The deregulation of alternative splicing (AS) has recently been implicated as a relevant source of molecular heterogeneity in cancer. However, the targets and intrinsic mechanisms on regulatory network of splicing in hepatocarcinogenesis are largely unknown. Here, we report a functional impact of the splice variant of Splicing Regulatory Glutamic Acid and Lysine Rich Protein 1 (SREK1) and its upstream regulator, Serine/arginine-rich splicing factor 10 (SRSF10) on sustaining the oncogenic signal. Overall design: To identify the alternative spliced targets of SRSF10 in HCC cells, we did the stable knockdown for the SRSF10 (3B-2, LM3-2) and Scramble control (3B-1, LM3-1) in Hep3B and HCCLM3 cells by lentivirus-mediated shRNA delivery. The RNA was extracted and sent for mRNA sequencing to identify the splicing targets regulated by SRSF10.

可变剪接（alternative splicing, AS）的失调近来被证实为癌症分子异质性的重要来源之一。然而，肝癌发生过程中剪接调控网络的作用靶点与内在机制仍尚不明确。本研究阐明了剪接调控富谷氨酸富赖氨酸蛋白1（Splicing Regulatory Glutamic Acid and Lysine Rich Protein 1, SREK1）的剪接变体及其上游调控因子丝氨酸/精氨酸富集剪接因子10（Serine/arginine-rich splicing factor 10, SRSF10）在维持致癌信号中的功能影响。
总体实验设计：为鉴定SRSF10在肝细胞癌（hepatocellular carcinoma, HCC）细胞中的可变剪接靶点，我们通过慢病毒介导的短发夹RNA（short hairpin RNA, shRNA）递送系统，在Hep3B与HCCLM3细胞中分别构建了SRSF10稳定敲低株（3B-2、LM3-2）及阴性对照株（3B-1、LM3-1）。随后提取细胞总RNA并进行mRNA测序，以鉴定受SRSF10调控的剪接靶点。