首页|肿瘤相关巨噬细胞PKM2表达对食管癌细胞恶性表型的影响

肿瘤相关巨噬细胞PKM2表达对食管癌细胞恶性表型的影响

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目的 探讨肿瘤相关巨噬细胞(Tumor associated macrophages,TAMs)中M2型丙酮酸激酶(Pyruvate kinase M2,PKM2)表达对食管癌细胞迁移、侵袭恶性表型的影响.方法 慢病毒转染PKM2敲低/过表达质粒至THP-1细胞并诱导分化为TAMs,随机分为PKM2敲低组(PKM2 KD组)、PKM2敲低对照组(PKM2 KD NC组)、PKM2过表达组(PKM2 OE组)和PKM2过表达对照组(PKM2 OE NC组).qRT-PCR检测PKM2及巨噬细胞表型标志物的表达;进一步与食管癌KYSE150细胞共培养后,Transwell实验检测食管癌细胞迁移和侵袭能力变化.将共培养细胞混合接种裸鼠皮下构建裸鼠荷瘤模型,免疫组化检测PKM2和CD163的表达,ELISA检测葡萄糖和乳酸水平.结果 与PKM2 KD NC组相比,PKM2 KD组TAMs细胞内PKM2表达水平显著降低,NOS2表达上调(P均<0.05),CD163轻微下调(P>0.05),CCL5表达下调(P<0.05);与PKM2 OE NC组相比,PKM2 OE组TAMs细胞内PKM2表达水平显著升高,NOS2表达下调(P均<0.05),CD163轻微上调(P>0.05),CCL5表达上调(P<0.05).共培养后,与PKM2 KD NC组相比,PKM2 KD组KYSE150细胞迁移和侵袭能力均显著下降(P均<0.001);与PKM2 OE NC组相比,PKM2 OE组KYSE150细胞迁移和侵袭能力均显著增强(P均<0.01).细胞混合接种后,与CO-PKM2 KD NC组相比,CO-PKM2 KD组裸鼠皮下移植瘤体积和重量均减小(P>0.05),瘤体内PKM2和CD163表达下降(P均>0.05),血清葡萄糖和乳酸水平显著下降(P均<0.05);与CO-PKM2 OE NC组相比,CO-PKM2 OE组裸鼠皮下移植瘤体积和重量均增大(P>0.05),瘤体内PKM2和CD163表达上调(P均>0.05),血清葡萄糖和乳酸水平显著升高(P均<0.05).结论 PKM2表达变化影响TAMs细胞表型分化,而TAMs细胞中PKM2的表达可能通过无氧糖酵解作用,影响食管癌细胞的恶性表型.
Effect of M2 pyruvate kinase(PKM2)expression in tumor associated macrophages(TAMs)on malignant phenotype of esophageal squamous cell carcinoma(ESCC)
Objective To investigate the effect of M2 pyruvate kinase(PKM2)expression in tumor associ-ated macrophages(TAMs)on the malignant phenotype of esophageal squamous cell carcinoma(ESCC)cell migration and invasion.Methods Transfection of PKM2 knockdown/overexpression plasmid with lentivirus into THP-1 cells,inducing THP-1 cells to differentiate into TAMs,which were randomly divided into PKM2 knockdown group(PKM2 KD group),PKM2 knockdown control group(PKM2 KD NC group),PKM2 overexpression group(PKM2 OE group)and PKM2 overexpression control group(PKM2 OE NC group).The expression of PKM2 and macrophage phenotype markers were detected by qRT-PCR.After cocultured with KYSE150 cells,transwell assay was used to detect the migration and invasion ability of ESCC cells.Cocultured cells were inoculated subcutaneously into nude mice to construct a nude mouse tumor bearing model.IHC was used to detect the expression of PKM2 and CD163,and ELISA was used to detect glucose and lactate levels.Results Compared with PKM2 KD NC group,the expression level of PKM2 was significantly reduced in TAMs cells in PKM2 KD group,while the expression of NOS2 was up-regulated(all P<0.05),the expression of CD163 was slightly downregulated(P>0.05),and the expres-sion of CCL5 was downregulated(P<0.05);Compared with PKM2 OE NC group,the expression level of PKM2 was significantly increased in TAMs cells in PKM2 OE group,while the expression of NOS2 was downregulated(all P<0.05),the expression of CD163 was slightly upregulated(P>0.05),and the ex-pression of CCL5 was upregulated(P<0.05);Compared with PKM2 KD NC group,the ability of migra-tion and invasion of KYSE150 cells after co-culture were significantly reduced in PKM2 KD group(all P<0.001).Compared with PKM2 OE NC group,the ability of migration and invasion of KYSE150 cells after co-culture were significantly enhanced in PKM2 OE group(all P<0.01).Compared with CO-PKM2 KD NC group,CO-PKM2 KD group showed a decrease in the volume and weight of subcutaneous transplanted tumors in nude mice with mixed cell inoculation(P>0.05).Meanwhile,the expression of PKM2 and CD163 was decreased in tumor tissue(P>0.05),and the concentration level of glucose and lactate in ser-um was significant decreased(all P<0.05).Compared with CO-PKM2 OE NC group,CO-PKM2 OE group showed an increase in the volume and weight of subcutaneous transplanted tumors in nude mice(P>0.05).The expression of PKM2 and CD163 was upregulated in tumor tissue(all P>0.05),and serum glucose and lactate levels were significantly elevated(all P<0.05).Conclusion Variation in PKM2 ex-pression affect the phenotypic differentiation of TAMs cells,and PKM2 expression in TAMs cells may af-fect the malignant phenotype of ESCC cells through anaerobic glycolysis.

esophageal squamous cell carcinoma(ESCC)tumor associated macrophages(TAMs)pyru-vate kinase M2(PKM2)malignant phenotype

刘清、谭依依、陈娇、彭天元、王薇、卢晓梅

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新疆医科大学第一附属医院临床医学研究院,省部共建中亚高发病成因与防治国家重点实验室,乌鲁木齐 830011

食管癌 肿瘤相关巨噬细胞 M2型丙酮酸激酶 恶性表型

新疆维吾尔自治区自然科学基金杰出青年科学基金项目省部共建中亚高发病成因与防治国家重点实验室开放课题项目

2021D01E31SKL-HIDCA-2020-SG2

2024

新疆医科大学学报
新疆医科大学

新疆医科大学学报

CSTPCD
影响因子:0.76
ISSN:1009-5551
年,卷(期):2024.47(2)
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