RNPS1通过Notch信号通路参与调控胰腺癌细胞的生存及进展

RNA-binding protein with serine-rich domain 1 regulates the survival and progression of pancreatic cancer cells through the Notch signaling pathway

摘要

目的 探讨富含丝氨酸结构域1的RNA结合蛋白（RNA-binding protein with serine-rich domain 1，RNPS1）在胰腺癌进展中的作用及可能分子机制。 方法 免疫组织化学与免疫荧光检测RNPS1与Notch3在胰腺癌组织及癌旁组织的表达；RT-qPCR、免疫荧光检测RNPS1与Notch3在胰腺癌细胞中的表达情况；Hoechst与CCK-8实验检测胰腺癌细胞凋亡与增殖；划痕实验与transwell实验检测胰腺癌细胞迁移与侵袭能力；Western blot实验检测胰腺癌细胞中N-Cadherin和E-Cadherin的表达；Western blot与RT-qPCR实验检测胰腺癌细胞中Notch3与HEY1的表达。 结果 与癌旁组织与正常细胞系相比较，RNPS1与Notch3在胰腺癌组织中及胰腺癌细胞的表达均增高（F=121.612、34.649，均P<0.05）；与对照组相比较，敲低RNPS1抑制生物标志物N-Cadherin的表达（t=39.922，P<0.05），促进E-Cadherin的表达（t=8.281，P<0.05），敲低RNPS1可减弱癌细胞的生存、迁移侵袭的能力（t=2.017、4.874、19.747，均P<0.05，），促进了细胞凋亡（t=33.673，P<0.05）；敲低RNPS1降低了癌细胞中Notch3与HEY1的表达（t=17.546、6.258，均P<0.05）。 结论 RNPS1的表达与胰腺癌细胞生存、恶性表型有关，RNPS1可能通过调控Notch3/HEY1信号通路促进胰腺癌细胞的生存及肿瘤进展。

Abstract

Objective To investigate the role and possible molecular mechanism of RNA-binding protein with serine-rich domain 1 （RNPS1） in the progression of pancreatic cancer. Methods Immunohistochemistry and immunofluorescence assay were used to measure the expression of RNPS1 and Notch3 in pancreatic cancer tissue and paracancerous tissue，and RT-qPCR and immunofluorescence assay were used to measure the expression of RNPS1 and Notch3 in pancreatic cancer cells. Hoechst staining and CCK-8 assay were used to measure the apoptosis and proliferation of pancreatic cancer cells； Wound healing assay and Transwell assay were used to observe the migration and invasion abilities of pancreatic cancer cells； Western blot was used to measure the expression of N-Cadherin and E-Cadherin in pancreatic cancer cells，and Western blot and RT-qPCR were used to measure the expression of Notch3 and HEY1 in pancreatic cancer cells. Results There were significant increases in the expression levels of RNPS1 and Notch3 in pancreatic cancer tissue and pancreatic cancer cells compared with the paracancerous tissue and normal cell lines（F=121.612 and 34.649，all P<0.05）. Comparing with the control group，RNPS1 knockdown inhibited the expression of the biomarker N-Cadherin（t=39.922，P<0.05），promoted the expression of E-Cadherin（t=8.281，P<0.05），reduced the survival，migration，and invasion abilities of cancer cells（t=2.017，4.874，and 19.747，all P<0.05），and promoted cancer cell apoptosis（t=33.673，P<0.05）. In addition，RNPS1 knockdown reduced the expression levels of Notch3 and HEY1 in cancer cells（t=17.546 and 6.258，all P<0.05）. Conclusion The expression of RNPS1 is associated with the survival and malignant phenotype of pancreatic cancer cells，and RNPS1 may promote the survival of pancreatic cancer cells and the progression of pancreatic cancer by regulating the Notch3/HEY1 signaling pathway.

关键词

选择性剪切 / 胰腺癌 / 富含丝氨酸结构域1的RNA结合蛋白 / 生存 / 进展

Key words

alternative splicing / pancreatic cancer / RNA-binding protein with serine-rich domain 1 / survival / progression

基金资助

新乡医学院博士科研启动基金资助项目(ZZ201815)