牙龈卟啉单胞菌上调Src蛋白表达对食管鳞癌细胞增殖、迁移和侵袭的影响  

Effect of Porphyromonas gingivalis up-regulating Src protein expression on proliferation,migration and invasion of esophageal squamous cell carcinoma

作　　者：郭一君 高社干[1] 石林林 GUO Yijun;GAO Shegan;SHI Linlin(Department of Oncology,The First Affiliated Hospital of Henan University of Science and Technology,Luoyang,Henan 471000,China)

机构地区：[1]河南科技大学第一附属医院肿瘤科,河南洛阳471000

出　　处：《中国临床研究》2025年第1期77-81,共5页Chinese Journal of Clinical Research

基　　金：国家自然科学基金资助项目(81972571)。

摘　　要：目的 探讨牙龈卟啉单胞菌(P.gingivalis,Pg)可否通过上调原癌基因酪氨酸蛋白激酶(Src)影响食管鳞状细胞癌(鳞癌)细胞KYSE30的增殖和迁移。方法 将KYSE30细胞分为四组:未感染组(Pg^(-)组,不加Pg菌液感染)、抑制剂组(不加Pg菌液感染,加Src抑制剂PP2 5 nM作用24 h)、感染组(Pg^(+)组,仅加Pg菌液50μL感染细胞)和感染+抑制剂组(Pg^(+)抑制剂组,先加PP2 5 nM作用24 h后,再加Pg菌液50μL感染细胞)。Pg^(-)组及Pg^(+)组感染24 h后,免疫荧光法检测Src和Pg的表达;Pg^(+)组和Pg^(+)抑制剂组细胞分别在感染0、12、24和48 h后,蛋白印迹法检测Src和STAT3蛋白表达量;Pg感染24 h后,Transwell实验、CCK-8法、流式细胞仪分别检测四组细胞迁移和侵袭数、增殖活性及凋亡率。结果 与Pg^(-)组比较,Pg^(+)组Src和Pg荧光强度均显著升高(P<0.05)。Pg^(+)组感染Pg 12、24、48 h时细胞中Src和STAT3蛋白表达量均较感染0 h升高(P<0.05);Pg^(+)抑制剂组感染12、24和48 h细胞中Src和STAT3蛋白表达量均较感染0 h时升高(P<0.05);感染后同一时点,Pg^(+)抑制剂组细胞中Src和STAT3蛋白表达量均较Pg^(+)组降低(P<0.05)。与Pg^(-)组比较,抑制剂组迁移和侵袭细胞数减少,细胞增殖活性降低,细胞凋亡率升高(P<0.05);与Pg^(-)组比较,Pg^(+)组迁移和侵袭细胞数增多,细胞增殖活性升高,细胞凋亡率降低(P<0.05);与Pg^(+)组比较,Pg^(+)抑制剂组迁移和侵袭细胞数减少,细胞增殖活性降低,细胞凋亡率升高(P<0.05)。结论 Pg感染可促进食管鳞癌细胞中Src和STAT3表达,抑制Src可逆转Pg对食管鳞癌细胞迁移和侵袭的促进作用,进而降低细胞增殖活性,诱导细胞凋亡。Objective To investigate the effect of Porphyromonas gingivalis(P.gingivalis,Pg)on the proliferation and migration of esophageal squamous cell(KYSE30)through up-regulating proto-oncogene tyrosine protein kinase(Src).Methods KYSE30 cells were divided into four groups:no infection group(Pg^(-)group,without Pg bacterial solution),inhibitor group(without Pg bacterial solution,with Src inhibitor PP25 nM for 24 h),infection group(Pg^(+)group,with only 50μL of Pg bacterial solution to infect cells)and infection+inhibitor group(Pg^(+)inhibitor group,PP25 nM for 24 h,then 50μL of Pg solution for infection).Twenty-four hours after infection in the Pg^(-)group and Pg^(+)group,the expressions of Src and Pg were detected by immunofluorescence.The expressions of Src and STAT3 proteins in the Pg^(+)group and Pg^(+)inhibitor group were detected by Western blot at 0,12,24 and 48 h,respectively.Twenty-four hours after Pg infection,the migration and invasion number,proliferative activity and apoptosis rate of cells in the four groups were detected by Transwell assay,CCK-8 method and flow cytometry,respectively.Results Compared with Pg^(-)group,the fluorescence intensity of Src and Pg in Pg^(+)group significantly increased(P<0.05).The expression levels of Src and STAT3 protein in Pg^(+)group increased at 12,24 and 48 h of Pg infection compared with at 0 h(P<0.05).The expression levels of Src and STAT3 protein in Pg^(+)inhibitor group at 12 and 24 h of Pg infection were higher than those at 0 h(P<0.05).At the same time point after infection,the expression levels of Src and STAT3 protein in Pg^(+)inhibitor group were lower than those in Pg^(+)group(P<0.05).Compared with Pg^(-)group,the number of migrating and invading cells in inhibitor group decreased,cell proliferation activity decreased,and cell apoptosis rate increased(P<0.05).Compared with Pg^(-)group,the number of migratory and invasive cells in Pg^(+)group increased,cell proliferation activity increased,and cell apoptosis rate decreased(P<0.05).Compared with Pg^(+)group,the n

关 键 词：食管鳞状细胞癌 原癌基因酪氨酸蛋白激酶 增殖 迁移 侵袭 牙龈卟啉单胞菌 

分 类 号：R735.1[医药卫生—肿瘤]