PKM1调控小细胞肺癌自噬与神经内分泌标志物表达  

PKM1 Regulates the Expression of Autophagy and Neuroendocrine Markers in Small Cell Lung Cancer

作　　者：唐辰晨 金禹龙 赵沛妍 田琳 李慧[2] 杨长良 钟睿 柳菁菁[3] 马丽霞[3] 程颖[2,3] Chenchen TANG;Yulong JIN;Peiyan ZHAO;Lin TIAN;Hui LI;Changliang YANG;Rui ZHONG;Jingjing LIU;Lixia MA;Ying CHENG(Biobank,Jilin Cancer Hospital,Changchun 130012,China;Medical Oncology Translational Research Lab,Jilin Cancer Hospital,Changchun 130012,China;Department of Medical Oncology,Jilin Cancer Hospital,Changchun 130012,China)

机构地区：[1]吉林省肿瘤医院生物样本库,长春130012 [2]吉林省肿瘤医院肿瘤转化医学实验室,长春130012 [3]吉林省肿瘤医院肿瘤内科,长春130012

出　　处：《中国肺癌杂志》2024年第9期645-653,共9页Chinese Journal of Lung Cancer

基　　金：国家癌症中心攀登基金项目(No.NCC201907B01);吉林省卫生健康科技能力提升项目(No.2021JC094);吉林省中医药科技项目(No.2023073)资助。

摘　　要：背景与目的小细胞肺癌(small cell lung cancer,SCLC)恶性程度高,异质性强,被称为难治性肿瘤。免疫治疗改变了广泛期SCLC(extensive-disease SCLC,ED-SCLC)的治疗格局,但受益人群有限,因此,寻找新的治疗措施是目前SCLC亟待解决的临床问题。SCLC具有高度活跃的糖酵解代谢特征,而丙酮酸激酶M1(pyruvate kinase M1,PKM1)是糖酵解途径中重要的限速酶PK的同工酶之一。研究表明PKM1与自噬及药物敏感性有关,但PKM1如何调控SCLC药物敏感性及其作用机制尚不清楚。本研究旨在探讨PKM1在SCLC中的生物学功能,包括PKM1对SCLC的增殖、迁移、自噬、药物敏感性及神经内分泌(neuroendocrine,NE)相关标志物表达的影响。方法应用Western blot检测SCLC细胞中PKM1的表达水平;通过慢病毒稳定转染构建PKM1基因过表达的SCLC细胞株,MTT法检测细胞增殖能力及药物敏感性,Transwell实验测定细胞迁移能力,流式细胞术检测细胞自噬水平,Western blot检测NE相关蛋白的表达水平。结果不同的SCLC细胞系PKM1表达具有差异性,H1092中PKM1表达较低(P<0.01)。与对照组相比,PKM1过表达的H1092细胞虽然增殖水平无明显差异,但迁移能力增高(P<0.001),药物敏感性降低,并且自噬水平受到抑制(P<0.001)。此外过表达PKM1可上调非神经内分泌(non-neuroendocrine,non-NE)相关蛋白表达(P<0.01),降低NE相关蛋白表达(P<0.01)。结论PKM1在SCLC细胞系中具有差异表达,PKM1高表达不影响SCLC的细胞增殖,但影响其迁移。PKM1可能通过抑制自噬,调节NE标志物的表达,从而影响药物敏感性,研究结果为探索PKM1在SCLC中的作用提供了理论依据。Background and objective Small cell lung cancer(SCLC)is known as recalcitrant cancer with high malignancy and heterogeneity.Immunotherapy has changed the treatment pattern of extensive-disease SCLC(ED-SCLC),but the beneficiary population is limited.Therefore,exploring new therapeutic strategies is an urgent clinical problem to be solved for SCLC.SCLC is characterized by highly active glycolytic metabolism and pyruvate kinase M1(PKM1)is one of the isozymes of PK,an important rate-limiting enzyme in glycolysis pathway.Previous studies have shown that PKM1 is related to autophagy and drug sensitivity,however,how PKM1 regulates drug sensitivity in SCLC and its mechanism remain unclear.The aim of this study was to investigate the biological functions of PKM1 in SCLC,including its effects on proliferation,migration,autophagy,drug sensitivity,and expression of neuroendocrine(NE)-related markers in SCLC.Methods Western blotwas used to detect the expression level of PKM1 in SCLC cells.PKM1 gene-overexpressed SCLC cell lines were constructed by stable lentivirus transfection.Proliferation of cells and drug sensitivity were detected by MTT,and migration ability of cells was determined by Transwell.The level of autophagy was detected by flow cytometry.Western blot was used to determine the expression levels of NE-related proteins.Results PKM1 was differentially expressed among various SCLC cell lines,and was lower in H1092 cells(P<0.01).Compared with the control group,there was no significant difference in proliferation level of PKM1 overexpressing H1092 cell,but the migration ability was significantly increased(P<0.001),the drug sensitivity was re-duced,and the level of autophagy was inhibited(P<0.001).Additionally,overexpression of PKM1 could upregulate the expres-sion of non-neuroendocrine(non-NE)-related proteins(P<0.01)and decrease the expression of NE-related proteins(P<0.01).Conclusion PKM1 was differentially expressed in SCLC cell lines,and high expression of PKM1 did not affect the prolifera-tion,but affected the mi

关 键 词：肺肿瘤 PKM1 自噬 神经内分泌 药物敏感性 

分 类 号：R734.2[医药卫生—肿瘤]