摘要
目的探究缝隙连接结合蛋白2(GJB2)在胰腺癌相关成纤维细胞(CAF)的表达和对胰腺癌细胞侵袭和迁移的影响。方法收集河南省人民医院39例胰腺癌标本和其对应癌旁组织,培养原代CAF和正常胰腺成纤维细胞(NF)以及胰腺癌类器官。测序分析CAF与NF基因表达差异。免疫组化染色进行GJB2表达评分,根据病理结果将标本分为T分期>2组和T≤2组,N>0组和N=0组,有神经浸润组和无神经浸润组。采用t检验评估不同分组GJB2表达评分差异,免疫组化染色进行GJB2表达评分,根据病理结果将标本分为T分期>2组和T≤2组,N>0组和N=0组,有神经浸润组和无神经浸润组。使用t检验评估不同分组GJB2表达评分差异,分析GJB2在CAF中表达量与患者预后相关性。设计GJB2过表达序列并构建载体,转染NF,蛋白质印迹法(Western blot)检测转染后NF中GJB2表达量。将GJB2过表达组设为实验组(OE-NF),转染空载质粒NF设为对照组(OE-NC),分别与胰腺癌细胞、胰腺癌类器官共培养,Transwell实验和侵袭胶实验,采用t检验评估两组细胞影响下胰腺癌细胞的转移能力和侵袭能力差异。结果测序结果显示GJB2在CAF表达量高于NF(log2FoldChange:-5.37 pval:1.34e-04<0.05),免疫组织化学结果显示GJB2在T分期>2组表达量(11.310±0.221)较T≤2组(9.261±0.542)明显升高,差异有统计学意义(t=3.99,P<0.01)。N>0组表达量(10.950±0.249)较N=0的组(9.313±0.669)升高,差异有统计学意义(t=2.857,P<0.05)。神经浸润组(11.600±0.193)较无神经浸润组(9.625±0.442)升高,差异有统计学意义(t=4.563,P<0.01)。Kaplan-Meier生存分析结果显示GJB2高表达组患者3年总生存期明显低于低表达组[风险比(HR)=2.50(1.26~4.96),P<0.01]。Transwell迁移实验中OE-NF组穿出数量(20.200±2.417)多于OE-NC组(6.600±1.288,t=4.966,P<0.01),且Transwell侵袭实验中OE-NF组(343.80±19.440)引导肿瘤细胞穿出数量多于OE-NC组(66.380±3.914,t=13.99,P<0.01)。结论GJB2在CAF�
Objective To study the expression of gap junction binding protein 2(GJB2)in pancreatic cancer-associated fibroblasts(CAFs)and its effect on the invasion and metastasis of pancreatic cancer cells.Methods The primary CAFs,normal fibroblasts(NFs)and pancreatic organoids were cultured from 39 pancreatic cancer specimens and corresponding paracancerous tissues collected from Henan Provincial People’s Hospital.The gene expression differences in CAFs and NFs was analyzed by RNA Sequencing.Immunohistochemical staining was used to score GJB2 expression.According to the pathological results,the specimens were divided into T stage>2 group and T≤2 group,N>0 group and N=0 group,with and without nerve infiltration group.The difference in GJB2 expression scores in different groups was evaluated by t-test,and the correlation between GJB2 expression in CAFs and prognosis was analyzed.The overexpression sequence of GJB2 was designed and the vector was constructed.After NFs were transfected,the expression of GJB2 in OE-NF was detected by Western blotting.GJB2 overexpression group was set as the experimental group(OE-NF)and NFs as the control group(OE-NC).Results RNA Sequencing results showed that the expression of GJB2 in CAFs was higher than that in normal pancreatic fibroblasts(log2FoldChange:-5.37 pval:1.34e-04<0.05).Immunohistochemical results showed that the expression of GJB2 in pancreatic cancer tissue of the T stage>2 group(11.310±0.221)was significantly higher than that of the T≤2 group(9.261±0.542),with a statistically significant difference(t=3.99,P<0.01).The expression level in N>0 group(10.95±0.249)was significantly higher than that in N=0 group(9.313±0.669,t=2.857,P<0.05).The expression in the group with nerve infiltration(11.600±0.193)was significantly higher than that in the group without nerve infiltration(9.625±0.442,t=4.563,P<0.01).Kaplan-Meier survival analysis showed that the 3-year total survival time in patients with high GJB2 expression was significantly lower than that in patients with low GJB2
作者
刘佳音
刘传江
付强
罗乾坤
刘攀
张宏伟
Liu Jiayin;Liu Chuanjiang;Fu Qiang;Luo Qiankun;Liu Pan;Zhang Hongwei(Department of Hepatobiliary and Pancreatic Surgery,People’s Hospital of Zhengzhou University,Zhengzhou 450003,China;Department of Hepatobiliary and Pancreatic Surgery,Henan Provincial People’s Hospital,Zhengzhou 450003,China)
出处
《中华实验外科杂志》
CAS
北大核心
2023年第2期266-270,共5页
Chinese Journal of Experimental Surgery
基金
河南省科技攻关项目(SBGJ202102027)。
关键词
胰腺癌
癌症相关成纤维细胞
缝隙连接结合蛋白2
侵袭
Pancreatic cancer
Cancer-associated fibroblast
Gap junction binding protein 2
Cancer invasion
