PKM2促进肿瘤发展的作用机制研究

doi:10.19586/j.2095-2341.2021.0181

生物技术进展 ›› 2022, Vol. 12 ›› Issue (3): 411-418.DOI: 10.19586/j.2095-2341.2021.0181

PKM2促进肿瘤发展的作用机制研究

王彩虹¹(), 陈剑飞², 鞠久君², 朱雪洁², 王彩霞²()

^1.首都医科大学附属北京友谊医院，北京热带医学研究所，北京 100050
^2.滨州医学院中西医结合学院，烟台市肿瘤代谢中药药理重点实验室，山东烟台 264003

收稿日期:2021-11-12 接受日期:2021-12-03 出版日期:2022-05-25 发布日期:2022-05-26
通讯作者: 王彩霞
作者简介:王彩虹 E-mail：13520137104@163.com；
基金资助:
国家自然科学基金项目(82103071)

The Mechanisms of PKM2 Promoting Tumor Progression

Caihong WANG¹(), Jianfei CHEN², Jiujun JU², Xuejie ZHU², Caixia WANG²()

^1.Beijing Friendship Hospital，Capital Medical University，Beijing Institute of Tropical Medicine，Beijing 100050，China
^2.Yantai Key Laboratory of Pharmacology of Traditional Chinese Medicine in Tumor Metabolism，School of Integrated Traditional Chinese and Western Medicine，Binzhou Medical University，Shandong Yantai 264003，China

Received:2021-11-12 Accepted:2021-12-03 Online:2022-05-25 Published:2022-05-26
Contact: Caixia WANG

摘要/Abstract

摘要：

为了分析丙酮酸激酶M2型（pyruvate kinase M2，PKM2）在不同肿瘤中的表达情况及其与肿瘤患者临床预后的关系，并探索PKM2对肿瘤细胞增殖和迁移的影响及其作用机制，用TCGA数据库和免疫印迹分析了33种肿瘤中PKM2的表达情况，探索了PKM2与不同肿瘤患者预后的关系。在肺癌细胞系中过表达PKM2，利用CCK8和Transwell方法分析PKM2对肺癌细胞增殖和迁移能力的影响。利用免疫印迹检测不同肿瘤细胞中过表达和敲低PKM2对热休克蛋白90α（Hsp90α）分泌的影响以及上皮-间质转化（epithelial-mesenchgmal transition，EMT）相关蛋白的变化。TCGA数据分析显示，PKM2在包括乳腺癌、肺癌等15种肿瘤中高表达，且9种肿瘤中PKM2的高表达与肿瘤的预后具有显著相关性。在肺癌细胞中过表达PKM2后，肺癌细胞的增殖和迁移能力显著增强。过表达PKM2能够显著增加乳腺癌和肺癌中Hsp90α的分泌。敲低PKM2能够抑制N-钙黏蛋白（N-cadhesion）和波形蛋白（Vimentin）的表达，促进E-钙黏蛋白（E-cadhesion）的表达。研究结果表明，PKM2在多种肿瘤中高表达且与肿瘤预后显著相关，能够通过影响Hsp90α的分泌以及上皮-间质转化相关蛋白的表达从而促进肿瘤的进展。PKM2有望成为潜在的广谱肿瘤标志物和治疗靶点。

关键词: 丙酮酸激酶M2型, 肿瘤预后, 热休克蛋白90α, N-钙黏蛋白, E-钙黏蛋白

Abstract:

To investigate the expression of pyruvate kinase M2 （PKM2） in different tumor tissue and its relationship with the clinical prognosis of cancer patients， and to explore the effects of PKM2 on tumor cell proliferation and migration and its mechanism of action， TCGA database and immunoblotting were used to analyze the expression of PKM2 in 33 tumors. The relationship between PKM2 and the prognosis of different tumor patients was explored. PKM2 was overexpressed in lung cancer cell lines， and the effect of PKM2 on lung cancer cell proliferation and migration were explored by CCK8 and Transwell assay. The effect of PKM2 overexpression on heat shock protein 90 alpha （Hsp90α） secretion in different tumor cells was detected by immunoblotting. Key proteins in the EMT process were detected after PKM2 knockdown. PKM2 was found highly expressed in 15 tumors， including breast cancer and lung cancer， and the high expression of PKM2 in 9 tumors was significantly correlated with the prognosis of tumors. After the expression of PKM2， the proliferation and migration ability of lung cancer cells was significantly enhanced. Overexpression of PKM2 could significantly increase Hsp90α secretion in breast cancer and lung cancer. Knocking down PKM2 could inhibit the expression of N-cadherin and Vimentin and increase the expression of E-cadherin. Results showed that， PKM2 was highly expressed in a variety of tumors and was significantly correlated with tumor prognosis. PKM2 could promote tumor progression by affecting the secretion of Hsp90α and the levels of epithelial-mesenchymal transition （EMT） related proteins. PKM2 may be a potential broad-spectrum tumor marker and therapeutic target.

Key words: pyruvate kinase M2, tumor prognosis, heat shock protein 90 alpha, N-cadherin, E-cadherin

中图分类号:

R730.2

王彩虹, 陈剑飞, 鞠久君, 朱雪洁, 王彩霞. PKM2促进肿瘤发展的作用机制研究[J]. 生物技术进展, 2022, 12(3): 411-418.

Caihong WANG, Jianfei CHEN, Jiujun JU, Xuejie ZHU, Caixia WANG. The Mechanisms of PKM2 Promoting Tumor Progression[J]. Current Biotechnology, 2022, 12(3): 411-418.

图/表 6

表1 TCGA数据库中肿瘤中英文名称及缩写

Table 1 The name and abbreviation of tumor in TCGA database

编号	名称	编号	名称
ACC	肾上腺皮质癌（adrenocortical carcinoma）	LUSC	肺鳞癌（lung squamous cell carcinoma）
BLCA	膀胱尿路上皮癌（bladder urothelial carcinoma）	MESO	间皮瘤（mesothelioma）
BRCA	乳腺浸润癌（breast invasive carcinoma）	OV	卵巢浆液性囊腺癌（ovarian serous cystadenocarcinoma）
CESC	宫颈鳞癌和腺癌（cervical squamous cell carcinoma and endocervical adenocarcinoma）	PAAD	胰腺癌（pancreatic adenocarcinoma）
CHOL	胆管癌（cholangio carcinoma）	PCPG	嗜铬细胞瘤和副神经节瘤（pheochromocytoma and paraganglioma）
COAD	结肠癌（colon adenocarcinoma）	PRAD	前列腺癌（prostate adenocarcinoma）
DLBC	弥漫性大B细胞淋巴瘤（lymphoid neoplasm diffuse large B-cell lymphoma）	READ	直肠腺癌（rectum adenocarcinoma）
ESCA	食管癌（esophageal carcinoma）	SARC	肉瘤（sarcoma）
GBM	多形成性胶质细胞瘤（glioblastoma multiforme）	SKCM	皮肤黑色素瘤（skin cutaneous melanoma）
HNSC	头颈鳞状细胞癌（head and neck squamous cell carcinoma）	STAD	胃癌（stomach adenocarcinoma）
KICH	肾嫌色细胞癌（kidney chromophobe）	TGCT	睾丸癌（testicular germ cell tumors）
KIRC	肾透明细胞癌（kidney renal clear cell carcinoma）	THCA	甲状腺癌（thyroid carcinoma）
KIRP	肾乳头状细胞癌（idney renal papillary cell carcinoma）	THYM	胸腺癌（thymoma）
LAML	急性髓细胞样白血病（acute myeloid leukemia）	UCEC	子宫内膜癌（uterine corpus endometrial carcinoma）
LGG	脑低级别胶质瘤（brain lower grade glioma）	UCS	子宫肉瘤（uterine carcinosarcoma）
LIHC	肝细胞肝癌（liver hepatocellular carcinoma）	UVM	葡萄膜黑色素瘤（uveal melanoma）
LUAD	肺腺癌（lung adenocarcinoma）

图1 PKM2在不同肿瘤中的表达情况A:TCGA数据库分析PKM基因在33种肿瘤及癌旁正常组织的表达情况，红色标记的瘤种差异具有统计学意义（P<0.05）；T—肿瘤组织，N—正常组织。TPM（transcript per million）表示样本中平均每一百万个转录本中对应于特定基因或转录本的数量；ACC—肾上腺皮质癌；BLCA—尿路上皮癌；BRCA—乳腺浸润癌；CESC—宫颈鳞癌和腺癌；CHOL—胆管癌；COAD—结肠癌；DLBC—弥漫性大B细胞淋巴瘤；ESCA—食管癌；GBM—多形成性胶质细胞瘤；HNSC—头颈鳞状细胞癌；KICH—肾嫌色细胞癌；KIRC—肾透明细胞癌；KIRP—肾乳头状细胞癌；LAML—急性髓细胞样白血病；LGG—脑低级别胶质瘤；LIHC—肝细胞肝癌；LUAD—肺腺癌；LUSC—肺鳞癌；MESO—间皮瘤；OV—卵巢浆液性囊腺癌；PAAD—胰腺癌；PCPG—嗜铬细胞瘤和副神经节瘤；PRAD—前列腺癌；READ—直肠腺癌；SARC—肉瘤；SKCM—皮肤黑色素瘤；STAD—胃癌；TGCT—睾丸癌；THCA—甲状腺癌；THYM—胸腺癌；UCEC—子宫内膜癌；UCS—子宫肉瘤；UVM—葡萄膜黑色素瘤。B：Western blotting检测PKM2在不同恶性程度的小鼠黑色素瘤细胞F0和F10中的表达，Actin为内参。

Fig. 1 The expression of PKM2 in different tumors

图2 PKM2的表达与不同肿瘤患者预后的关系注：利用TCGA数据库分析不同肿瘤中PKM2表达与患者预后的关系，CESC—宫颈鳞癌和腺癌，HNSC—头颈鳞状细胞癌，KIRC—肾透明细胞癌，LAML—急性髓细胞样白血病，LIHC—肝细胞肝癌，LUAD—肺腺癌，MESO—间皮瘤，PAAD—胰腺癌，UVM—葡萄膜黑色素瘤。

Fig. 2 The relationship of expression of PKM2 and the prognosis of different tumor patients

图3 过表达PKM2对肺癌细胞增殖和迁移的影响A：Western blotting检测PKM2在A549细胞中过表达效率；B：过表达PKM2对肺癌细胞增殖能力的影响；C：过表达PKM2对肺癌迁移能力影响的代表图像；D：过表达PKM2对肺癌迁移能力影响的定量结果。***表示与CT组相比，PKM2组的差异具有统计学意义（P<0.000 1）。

Fig. 3 The effect of PKM2 overexpression on the proliferation and migration of lung cancer cells

图4 过表达PKM2对Hsp90α分泌的影响A: Western blotting 检测过表达PKM2后，乳腺癌MCF-7细胞胞内和胞外Hsp90α的变化；B:Western blotting 检测过表达PKM2后，肺癌A549细胞胞内和胞外Hsp90α的变化。TCL表示全细胞裂解液，CM表示细胞培养上清。Co.St.表示考马斯亮蓝染色，作为细胞培养上清的上样对照。

Fig. 4 The effect of PKM2 overexpression on the secretion of Hsp90α

图5 敲低PKM2对EMT的影响

Fig. 5 Effect of PKM2 knockdown on EMT

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PKM2促进肿瘤发展的作用机制研究

The Mechanisms of PKM2 Promoting Tumor Progression

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