Wu Jing, Liu Gaifang, An Kang, Shi Linping
Department of Digestion, Hebei General Hospital, Shijiazhuang, Hebei 050051, P.R. China.
Oncol Lett. 2022 May;23(5):154. doi: 10.3892/ol.2022.13275. Epub 2022 Mar 16.
Pancreatic cancer (PC), one of the deadliest diseases worldwide, has exhibited an increasing incidence rate in recent years. The present study aimed to explore the biological mechanism of PC. Therefore, the expression levels of neuronal pentraxin 1 (NPTX1) and RNA-binding protein 10 (RBM10) were detected in PC cell lines using reverse transcription-quantitative PCR (RT-qPCR) and western blot analyses prior to or following NPTX1 and RBM10 overexpression. Additionally, the proliferative ability of PANC-1 and BxPC-3 cells treated with or without gemcitabine (GEM) and cisplatin (DDP) was evaluated using Cell Counting Kit-8 assay. Cell apoptosis and the expression levels of apoptosis-related proteins were determined by TUNEL assay and western blot analysis, respectively. Furthermore, wound healing and Transwell assays were performed to measure the migration and invasion abilities of PANC-1 and BxPC-3 cells. The interaction between RBM10 and NPTX1 mRNA was detected by RNA binding protein immunoprecipitation (RIP) assay. Additionally, cells were treated with actinomycin D to verify the regulatory effect of RBM10 on NPTX1 expression. This effect was further confirmed by RT-qPCR analysis. The results showed that NPTX1 was downregulated in PC cell lines. In addition, NPTX1 overexpression inhibited the proliferation and promoted apoptosis in PC cells. The results from the wound healing and Transwell assays revealed that the migration and invasion abilities of PANC-1 and BxPC-3 cells were reduced following NPTX1 overexpression. However, treatment of NPTX1-overexpressing cells with GEM or DDP attenuated PC cell viability. In addition, the results of the RIP assay revealed that RBM10 could bind with NPTX1. Furthermore, RBM10 overexpression could regulate NPTX1 expression, as evidenced by actinomycin D experiments. Overall, the results of the present study suggested that NPTX1 could inhibit PC and enhance the sensitivity of PC cells to chemotherapy. Additionally, NPTX1 was found to interact with RBM10, indicating that NPTX1 could inhibit PC via targeting RBM10.
胰腺癌（PC）是全球最致命的疾病之一，近年来其发病率呈上升趋势。本研究旨在探索胰腺癌的生物学机制。因此，在过表达神经元五聚体蛋白1（NPTX1）和RNA结合蛋白10（RBM10）之前或之后，使用逆转录定量PCR（RT-qPCR）和蛋白质免疫印迹分析检测PC细胞系中NPTX1和RBM10的表达水平。此外，使用细胞计数试剂盒-8检测法评估吉西他滨（GEM）和顺铂（DDP）处理或未处理的PANC-1和BxPC-3细胞的增殖能力。分别通过TUNEL检测法和蛋白质免疫印迹分析确定细胞凋亡及凋亡相关蛋白的表达水平。此外，进行伤口愈合实验和Transwell实验以检测PANC-1和BxPC-3细胞的迁移和侵袭能力。通过RNA结合蛋白免疫沉淀（RIP）实验检测RBM10与NPTX1 mRNA之间的相互作用。此外，用放线菌素D处理细胞以验证RBM10对NPTX1表达的调节作用。RT-qPCR分析进一步证实了这种作用。结果显示，PC细胞系中NPTX1表达下调。此外，NPTX1过表达抑制PC细胞增殖并促进其凋亡。伤口愈合实验和Transwell实验结果显示，NPTX1过表达后，PANC-1和BxPC-3细胞的迁移和侵袭能力降低。然而，用GEM或DDP处理过表达NPTX1的细胞会减弱PC细胞活力。此外，RIP实验结果显示RBM10可与NPTX1结合。此外，放线菌素D实验证明，RBM10过表达可调节NPTX1表达。总体而言，本研究结果表明NPTX1可抑制胰腺癌并增强PC细胞对化疗的敏感性。此外，发现NPTX1与RBM10相互作用，表明NPTX1可通过靶向RBM10抑制胰腺癌。
