lncCASC2通过miR-222-3p/CDK19轴调控胃癌NK细胞杀伤敏感性  

lncCASC2 regulates NK cell killing sensitivity in gastric cancer by targeting miR-222-3p/CDK19

作　　者：郭伟胜 付利然[2] 赵林 魏光亚 张娟 GUO Weisheng;FU Liran;ZHAO Lin;WEI Guangya;ZHANG Juan(Henan Provincial Hospital of Traditional Chinese Medicine,the Second Affiliated Hospital of Henan University of Traditional Chinese Medicine,General Surgery Second District,Zhengzhou 450000,China)

机构地区：[1]河南省中医院,河南中医药大学第二附属医院普外二区,郑州450000 [2]郑州人民医院中医科,郑州450000 [3]河南省中医院,河南中医药大学第二附属医院肿瘤内科,郑州450002

出　　处：《中国免疫学杂志》2022年第19期2352-2357,共6页Chinese Journal of Immunology

基　　金：国家自然科学基金项目(81303096)。

摘　　要：目的:探究CDK19对胃癌NK细胞杀伤敏感性的影响及作用机制。方法:TCGA数据库分析胃癌患者及细胞系中CDK19表达及其与生存期的关联。CCK-8法检测敲低CDK19后MGC-803、NCI-N87细胞增殖能力,流式细胞术检测细胞凋亡。LDH法检测sh-CDK19、sh-CASC2、miR-222-3p inhibitor、IL-6、IL-8、anti-IL-6、anti-IL-8对胃癌NK细胞杀伤敏感性的影响。Antibody Chip检测敲低CDK19后NCI-N87细胞中细胞因子表达。qPCR检测CDK19、IL-6、IL-8 mRNA表达,Western blot检测CDK19蛋白表达,双荧光素酶报告基因检测miR-222-3p和CDK19的靶向关系。结果:CDK19在胃癌患者及细胞系中高表达。低表达CDK19的胃癌患者总生存期更长,在MGC-803、NCI-N87细胞中敲低CDK19,可抑制细胞增殖,促进细胞凋亡,抑制IL-6、IL-8表达,增强NK细胞杀伤敏感性。NCI-N87细胞中lncCASC2通过miR-222-3p正向调控CDK19表达,进而抑制NK细胞杀伤敏感性。结论:胃癌细胞中CDK19呈高表达,受lncCASC2/miR-222-3p调控,促进细胞增殖,抑制细胞凋亡,且促进IL-6、IL-8表达,抑制胃癌NK细胞杀伤敏感性。Objective:To explore effect and mechanism of CDK19 on NK cell killing sensitivity in gastric cancer.Methods:TCGA database was used to analyze expression of CDK19 in gastric cancer patients and cell lines and its association with overall survival rate.CCK-8 assay was carried out to detect proliferation of MGC-803 and NCI-N87 cells after CDK19 knockdown,and flow cytometry was used to detect cells apoptosis.LDH method was used to detect effects of sh-CDK19,sh-CASC2,miR-222-3p inhibitor,IL-6,IL-8,anti-IL-6 and anti-IL-8 on NK cell killing sensitivity of gastric cancer.Cytokines expressions in NCI-N87 cells after CDK19 knockdown were detected by Antibody Chip.qPCR was used to detect mRNA expressions of CDK19,IL-6 and IL-8.CDK19protein expression was detected by Western blot,and dual luciferase reporter gene was used to detect target relationship between miR-222-3p and CDK19.Results:CDK19 was highly expressed in gastric cancer patients and cell lines.Knockdown of CDK19 in MGC-803 and NCI-N87 cells could inhibit cell proliferation,promote cell apoptosis,inhibit expressions of IL-6 and IL-8,and enhance killing sensitivity of NK cells.In NCI-N87 cells,lncCASC2 positively regulated CDK19 expression through miR-222-3p,then inhibited NK cell killing sensitivity.Conclusion:CDK19 is highly expressed in gastric cancer cells,which is regulated by lncCASC2/miR-222-3p,promotes cell proliferation,inhibits cell apoptosis,and promotes expressions of IL-6 and IL-8,thereby inhibiting cytotoxicity of NK cells.

关 键 词：胃癌 lncCASC2 miR-222-3p CDK19 NK细胞 

分 类 号：R735.2[医药卫生—肿瘤]