ABCA1与PAK4相互作用调节胃肠道间质瘤细胞生物学功能研究  

ABCA1 interacts with PAK4:implications for biological function of gastrointestinal stromal tumors cells

作　　者：高福来[1] 李莉[1] 谢长顺[1] 刘烨 张晓宇 邹春燕[1] GAO Fulai;LI Li;XIE Changshun;LIU Ye;ZHANG Xiaoyu;ZOU Chunyan(Department of Gastroenterology,The First Hospital of Qinhuangdao,Qinhuangdao,Hebei 066000,China)

机构地区：[1]秦皇岛市第一医院消化内科,河北秦皇岛066000

出　　处：《安徽医药》2025年第4期759-763,I0004,共6页Anhui Medical and Pharmaceutical Journal

基　　金：河北省医学科学研究课题项目(20221609)。

摘　　要：目的探讨ATP结合盒转运体A1(ABCA1)和p21激活激酶4(PAK4)的相互作用及其对胃肠道间质瘤细胞系(GIST-T1)生物学的影响。方法选取秦皇岛市第一医院在2021年3月至2022年11月手术切除的胃肠道间质瘤病人的肿瘤组织和正常胃黏膜组织,免疫组化和免疫荧光检测ABCA1和PAK4在胃肠道间质瘤病人肿瘤组织和正常胃黏膜组织表达;实时荧光定量逆转录聚合酶链反应(qRT-PCR)检测ABCA1和PAK4在胃肠道间质瘤细胞系GIST-T1和人正常肠上皮细胞NCM-460中表达;qRT-PCR确定ABCA1和PAK4在GIST-T1的相互作用;免疫荧光确定ABCA1和PAK4在GIST-T1内的表达;CCK-8研究ABCA1和PAK4对GISTT1细胞增殖能力影响;Trans-well小室探究过表达ABCA1和PAK4对GIST-T1细胞迁移和侵袭能力影响;划痕愈合观察ABCA1和PAK4对GIST-T1细胞划痕愈合能力影响。结果ABCA1在肿瘤组织(29.15±3.98)中染色阳性面积显著高于正常胃黏膜组织(3.89±0.87);PAK4在肿瘤组织(16.90±2.85)中染色阳性面积显著高于正常胃黏膜组织(3.95±0.73);ABCA1在GIST-T1的阳性细胞数(48.00±6.35)显著高于NCM-460(13.070±5.38);PAK4在GIST-T1的阳性细胞数(42.00±5.03)显著高于NCM-460(13.67±3.74);ABCA1过表达细胞株细胞48h后增殖能力(1.09±0.03)显著高于空白对照组细胞增殖能力(0.75±0.01);PAK4过表达细胞株细胞48h后增殖能力(1.03±0.06)显著高于空白对照组细胞增殖能力(0.62±0.04);PAK4随着ABCA1过表达而增加(pcDNA3.1-NC比pcDNA3.1-ABCA1为1.03±0.01比3.63±0.74);ABCA1随着PAK4过表达而增加(pcDNA3.1-NC比pcDNA3.1-PAK4为0.98±0.01比2.52±0.03)。结论ABCA1和PAK4相互作用促进胃肠道间质瘤细胞的恶性生物学行为。Objective To explore the interaction between ATP binding cassette transporter A1(ABCA1)and p21 protein activated kinase 4(PAK4)and its implications for the biology of the gastrointestinal stromal tumor cell line GIST-T1.Methods Tumor tissues from patients with gastrointestinal stromal tumors and normal gastric mucosa tissues,which were surgically resected in The First Hospital of Qinhuangdao between March 2021 and November 2022,were selected for the study.Immunohistochemical and immunofluorescent analyses were conducted to assess the expressions of ABCA1 and PAK4 in tumor tissues from patients with gastric stromal tumors,as well as in normal gastric mucosal tissues.Real-time fluorescent quantitative reverse transcription polymerase chain reaction(qRTPCR)was performed to assess the expressions of ABCA1 and PAK4 in GIST-T1 derived from gastric intestinal stromal tumor and the NCM-460 cell line derived from normal human colon epithelial cells.qRT-PCR confirmed the interaction between ABCA1 and PAK4 in GIST-T1.The expressions of ABCA1 and PAK4 within GIST-T1 was assessed using immunofluorescence analysis.The impact of ABCA1 and PAK4 on the proliferative capacity of GIST-T1 was studied using Cell Counting Kit-8(CCK-8)assay.The Trans-well chambers were utilized to explore the impact of ABCA1 and PAK4 overexpressions on the migratory and invasive capabilities of GIST-T1 cells.Enhanced wound healing capacity of GIST-T1 cells was observed in response to the influence of ABCA1 and PAK4.Results The staining positive area of ABCA1 in tumor tissues was significantly bigger than that in normal gastric mucosa tissues(29.15±3.98 vs.3.89±0.87).Similarly,the staining positive area of PAK4 in tumor tissues was significantly bigger than that in normal gastric mucosa tissues(16.90±2.85 vs.3.95±0.73).The positive cell count of ABCA1 in GIST-T1 was notably higher than that in NCM-460(48.00±6.35 vs.13.070±5.38),and the positive cell count of PAK4 in GIST-T1 was significantly greater than that in NCM-460(42.00±5.03 vs.13.67±3.74)

关 键 词：胃肠道间质肿瘤 HDL脂质转运蛋白ATP结合盒转运体A1 p21激活激酶4 增殖 迁移 侵袭 

分 类 号：R73[医药卫生—肿瘤]