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《天津医科大学学报》[ISSN:1006-8147/CN:12-1259/R]

卷:

31卷

期数:

2025年06期

页码:

505-513

栏目:

肿瘤疾病专题

出版日期:

2025-11-20

文章信息/Info

Title:

The role of RAI14 in malignant biological behaviors of pancreatic ductal adenocarcinoma cells and underlying molecular mechanisms

文章编号:

1006-8147(2025)06-0505-09

作者:

孙振兴; 赵 婷; 赵 樑

（天津医科大学肿瘤医院胰腺中心，国家恶性肿瘤临床医学研究中心，天津市恶性肿瘤临床医学研究中心，天津市消化系统肿瘤重点实验室，天津 300060）

Author(s):

SUN Zhenxing;  ZHAO Ting;  ZHAO Liang

（Pancreas Center， Tianjin Medical University Cancer Institute and Hospital， National Clinical Research Center for Cancer， Tianjin Key Laboratory of Digestive Cancer， Tianjin′s Clinical Research Center for Cancer， Tianjin 300060， China）

关键词:

胰腺癌; RAI14; 细胞增殖; PD-L1

Keywords:

pancreatic cancer;  RAI14;  cell proliferation;  PD-L1

分类号:

R735

DOI:

10.20135/j.issn.1006-8147.2025.06.0505

文献标志码:

A

摘要:

目的：探讨视黄酸诱导蛋白14（RAI14）对胰腺导管腺癌（PAAD）细胞恶性生物学行为的影响及其潜在分子机制。方法：利用基于基因表达水平值的交互分析平台（GEPIA）数据库分析RAI14在PAAD组织与正常胰腺组织中的mRNA差异表达；构建RAI14稳定敲低的人胰腺癌细胞系SW1990和PANC-1；利用Cell Counting Kit-8（CCK-8）实验、5-乙炔基-2′-脱氧尿苷（EDU）实验检测RAI14对细胞增殖的影响；通过Transwell迁移和侵袭实验、划痕实验评估细胞的迁移及侵袭能力；利用生物信息学分析和免疫组化实验检测RAI14与免疫检查点细胞程序性死亡-配体1（PD-L1）的相关性调控关系；Western印迹实验评估敲低RAI14组，Janus激酶/信号转导与转录激活子（JAK-STAT）通路与PD-L1的表达水平及调控关系。结果：生物信息学分析显示，PAAD组织中RAI14的mRNA水平显著高于正常组织（P<0.05）；与对照组相比，RAI14敲低的SW1990和PANC-1细胞增殖显著受抑：两种敲低RAI14细胞系CCK-8检测均显示在2～5 d细胞活力持续降低（FSW1990 =27.07、41.2、526.9、164.2，均P≤0.001；FPANC-1 =19.27、43.53、259.4、171.9，均P<0.005）；两种敲低RAI14细胞系EDU阳性细胞比例均显著降低（tSW1990 =30.41、21.32，均P<0.001；tPANC-1 =60.84、25.50，均P<0.001）；同时，与对照组相比，敲低RAI14的SW1990和PANC-1细胞迁移能力降低（tSW1990=7.555、7.816，均P<0.01；tPANC-1 =32.34、43.89，均P<0.01）、侵袭能力减弱（tSW1990 =22.5、16.34，均P<0.001；tPANC-1 =34.84、24.55，均P<0.001），24及48 h 细胞迁移率均显著下降（FSW1990 =112.9、261.0，均P<0.01；FPANC-1=285.8、184.3，均P<0.01）。免疫组化染色结果显示，高表达RAI14的胰腺癌组织呈现PD-L1高表达；低表达RAI14的胰腺癌组织则表现出PD-L1低表达。同时，RAI14和PD-L1在胰腺癌中存在很强的相关性（r=0.56，P=0.000）。Western印迹结果显示，与对照组相比，敲低RAI14的SW1990和PANC-1细胞中PD-L1、JAK1和STAT1的磷酸化水平显著降低。结论：RAI14促进胰腺癌细胞的增殖、迁移及侵袭。同时，RAI14通过JAK-STAT通路正向调控PD-L1表达，促进肿瘤的恶性行为。

Abstract:

Objective： To investigate the effect of retinoic acid-induced protein 14 （RAI14） on the malignant biological behaviors of pancreatic ductal adenocarcinoma （PAAD） cells and its underlying molecular mechanisms. Methods： Gene Expression Profiling Interactive Analysis （GEPIA） database was used to analyze the differential mRNA expression of RAI14 between PAAD tissues and normal pancreatic tissues. Stable RAI14 knockdown human pancreatic carcinoma lines SW1990 and PANC-1 were established. The effect of RAI14 on cell proliferation was assessed via cell counting kit-8 （CCK-8） and 5-ethynyl-2′-deoxyuridine （EDU） assays. The migration and invasion ability were evaluated through Transwell and scratch healing assays. The correlation and regulatory interactions between RAI14 and the immune checkpoint protein of programmed cell death ligand （PD-L1） were analyzed using bioinformatics tools and immunohistochemistry. Western blotting analysis was performed to evaluate the expression levels of the Janus kinase-signal transducer and activator of transcription（JAK-STAT） signaling pathway components and PD-L1， as well as their regulatory relationship， in RAI14 knockdown cells. Results： Compared with normal pancreatic tissue， bioinformatics analysis showed that RAI14 mRNA levels are significantly up-regulated in PAAD tissue （P<0.05）. Compared with the control group， RAI14 knockdown significantly inhibited the proliferation of SW1990 and PANC-1 cells. CCK-8 assays showed a continuous decrease in cell viability over 2-5 days in both RAI14 knockdown cell lines （FSW1990=27.07， 241.2， 526.9， 164.2， all P≤0.001； FPANC-1=19.27， 43.53， 259.4， 171.9， all P<0.005）. EDU assays demonstrated a significant reduction in the percentage of positive cells in two RAI14 knockdown cell lines （tSW1990=30.41， 21.32， both P<0.001； tPANC-1=60.84， 25.50， both P<0.001）； compared with the control group， RAI14 knockdown significantly decr-eased the migratory abilities of SW1990 and PANC-1 cells （tSW1990=7.555， 7.816， both P<0.01； tPANC-1=32.34， 43.89， both P<0.01）， as well as their invasive capabilities （tSW1990=22.5， 16.34， both P<0.001； tPANC-1=34.84， 24.55， both P<0.001）， and their migration rates at 24 and 48 hours （FSW1990=112.9， 261.0， both P<0.01； FPANC-1=285.8， 184.3， both P<0.01）. Additionally， immunohistoche-mical staining revealed that pancreatic cancer tissues with high RAI14 expression exhibited elevated PD-L1 levels， whereas tissues with low RAI14 expression showed reduced PD-L1 expression. Furthermore， a strong positive correlation was observed between RAI14 and PD-L1 expression in pancreatic cancer （r=0.56， P=0.000）. Western blotting indicated that silencing RAI14 in SW1990 and PANC-1 cells led to a marked decrease in the expression levels of PD-L1， as well as the phosphorylation of Janus kinase （JAK1） and signal transducer and activator of transcription （STAT1）， compared with control cells. Conclusion： RAI14 promotes the proliferation， migration， and invasion of pancreatic cancer cells. Moreover， RAI14 positively regulates PD-L1 expression via the JAK-STAT signaling pathway， thereby contributing to the malignant behavior of tumor cells.

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备注/Memo

备注/Memo:

基金项目 天津市医学重点学科（专科）建设项目（TJYXZDXK-009A）
作者简介 孙振兴（1998-），男，硕士在读，研究方向：蛋白质稳态与肿瘤代谢；通信作者：赵樑，E-mail：zhaoliangzl@tmu.edu.cn。

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更新日期/Last Update: 2025-11-20