LAPTM4B-35蛋白作为肝癌血清学诊断新标志物的探讨

doi:10.19723/j.issn.1671-167X.2021.04.015

摘要/Abstract

摘要：

目的: LAPTM4B-35是北京大学基础医学院发现和鉴定的一个肿瘤驱动基因LAPTM4B所编码的蛋白质同型分子(isoform)之一,其在多种恶性肿瘤中[如肝癌、肺癌(包括非小细胞肺癌和小细胞肺癌)、胃癌、结直肠癌、胰腺癌、胆囊癌、胆管癌、乳腺癌、前列腺癌、宫颈癌、子宫内膜癌等]均超高表达。实验室和临床资料均证明,LAPTM4B-35蛋白的过表达促进肿瘤生长、转移和多药耐药,LAPTM4B-35的蛋白表达水平与肝癌的复发相关。本文的目的在于鉴定患者体内的肝癌细胞和体外培养的肝癌细胞系是否释放出LAPTM4B-35蛋白到血液和细胞培养液中,以及其可能的存在形式,为建立肝细胞癌等肿瘤的血清学诊断新方法奠定基础。方法: 采用免疫印迹分析(Western blot)、酶联免疫吸附实验(enzyme-linked immunosorbent assay,ELISA)鉴定LAPTM4B-35蛋白,应用超滤和超离心方法从肝癌细胞培养上清液中分离、纯化外排体(exosome)。结果: 特异性抗体的ELISA结果表明,肝细胞癌患者的血液中存在LAPTM4B-35蛋白,体外培养的肝癌细胞以外排体形式释放LAPTM4B-35蛋白到培养基的上清液中。ELISA夹心法检测表明,肝细胞癌患者(n=43)血淸中LAPTM4B-35蛋白的平均水平和中位值均显著高于正常人(n=33)。结论: 肝癌细胞以外排体形式释放到其外环境中的LAPTM4B-35蛋白有望成为肝细胞癌血清学诊断的新标志物。

关键词: 肝细胞癌, 外排体, LAPTM4B-35, 生物标记, 肿瘤

Abstract:

Objective: LAPTM4B-35 protein is one of the isoforms that are encoded by a cancer driver gene, LAPTM4B. This gene was primarily found and identified in our lab of Peking University School of Basic Medical Sciences. The LAPTM4B-35 protein and its encoded mRNA are significantly over-expressed in a variety of cancers, such as hepatocellular carcinoma (HCC), lung cancers (including non small-cell lung cancer and small-cell lung cancer), stomach cancer, colorectal carcinoma, pancreatic cancer, gallbladder cancer, cholangiocarcinoma, breast cancer, prostate cancer, ovarian cancer, cervical cancer, endometrial cancer, and so on. It has firmly demonstrated through lab experiments either in vivo or in vitro, as well as clinical studies that the over-expression of LAPTM4B-35 can promote cancer growth, metastasis, and multidrug resistance. Specially, the expressive level of LAPTM4B-35 is associa-ted with recurrence of HCC. The aim of this study is to identify the release of LAPTM4B-35 protein from hepatocellular carcinoma into blood of HCC patients and into the medium of cultured HCC cells, and to identify its possible form of LAPTM4B-35 protein existed in blood and cell culture medium, as well as to explore the possibility of LAPTM4B-35 protein as a novel HCC biomarker for diagnosis of HCC and prognosis of HCC patients. Methods: Immunobloting (Western blot) and enzyme-linked immunosorbent assay (ELISA) were used for identification of LAPTM4B-35 protein in the blood of HCC patients and normal individuals. Ultrafiltration and ultracentrifugation were used to isolate and purify exosomes from the culture medium of HCC cells. Results: LAPTM4B-35 protein existed in the blood from HCC patients and normal donors that were demonstrated through Western blot and ELISA. LAPTM4B-35 was also released into the culture medium of HCC cells in the form of exosomes. Preliminary experiments showed that the average and the median of LAPTM4B-35 protein level in the blood of HCC patients (n=43) were both significantly higher than that in the blood of normal donors (n=33) through sandwich ELISA. Conclusion: It is promising that the LAPTM4B-35 protein which is released from HCC cells in the form of exosomes into their extraenvironment may be exploited as a novel cancer biomarker for HCC serological diagnosis.

Key words: Hepatocellular carcinoma, Exosomes, LAPTM4B-35, Biomarkers, tumor

中图分类号:

R735.7

庞泳,张沙,杨华,周柔丽. LAPTM4B-35蛋白作为肝癌血清学诊断新标志物的探讨[J]. 北京大学学报（医学版）, 2021, 53(4): 710-715.

PANG Yong,ZHANG Sha,YANG Hua,ZHOU Rou-li. Serum LAPTM4B-35 protein as a novel diagnostic marker for hepatocellular carcinoma[J]. Journal of Peking University (Health Sciences), 2021, 53(4): 710-715.

图/表 6

图1

图2

图3

图4

图5

图6

参考文献 21

[1]	Shao GZ, Zhou RL, Zhang QY, et al. Molecular cloning and characterization of LAPTM4B, a novel gene upregulated in hepatocellular carcinoma [J]. Oncogene, 2003, 22(32):5060-5069. doi: 10.1038/sj.onc.1206832
[2]	Yang H, Xiong FX, Lin M, et al. LAPTM4B-35 is a novel diagnostic marker and a prognostic factor of hepatocellular carcinoma [J]. J Surgical Oncology, 2010, 101(5):363-369. doi: 10.1002/jso.v101:5
[3]	Yang H, Xiong FX, Lin M, et al. LAPTM4B-35 overexpression is a risk factor for tumor recurrence and poor prognosis in hepatocellular carcinoma [J]. J Cancer Res Clin Oncol, 2010, 136(2):275-281. doi: 10.1007/s00432-009-0659-4 pmid: 19690886
[4]	何静, 邵根泽, 周柔丽, 等. 肝癌中高表达的新基因LAPTM4B对细胞增殖及成瘤性的影响 [J]. 北京大学学报(医学版), 2003, 35(4):348-352.
[5]	Li L, Shan Y, Yang H, et al. Upregulation of LAPTM4B-35 promotes malignant transformation and tumorigenesis in L02 human liver cell line [J]. Anat Rec (Hoboken), 2011, 294(7):1135-1142. doi: 10.1002/ar.v294.7
[6]	Liu XR, Xiong FX, Wei XH, et al. LAPTM4B-35, a novel tetratransmembrane protein and its PPRP motif play critical roles in proliferation and metastatic potential of HCC cells [J]. Cancer Sci, 2009, 100(12):2335-2340. doi: 10.1111/cas.2009.100.issue-12
[7]	Li L, Wei XH, Pan YP, et al. LAPTM4B: A novel cancer-asso-ciated gene motivates multi-drug resistance through efflux and activating PI3K/Akt signaling [J]. Oncogene, 2010, 29(43):5785-5795. doi: 10.1038/onc.2010.303 pmid: 20711237
[8]	Yang H, Xiong FX, Wei XH, et al. LAPTM4B-35 promotes growth and metastasis of hepatocelluar carcinoma [J]. Cancer Lett, 2010, 264(2):209-217. doi: 10.1016/j.canlet.2008.01.025
[9]	刘歆荣, 周柔丽, 张青云, 等. 人肝癌相关新基因编码产物LAPTM4B的鉴定及其生物学特性 [J]. 北京大学学报(医学版), 2003, 35(4):340-347.
[10]	Keller S, Sanderson MP, Stoeck A, et al. Exosomes: from biogenesis and secretion to biological function [J]. Immunol Lett, 2006, 107(2):102-108. doi: 10.1016/j.imlet.2006.09.005
[11]	Simons M, Raposo G. Exosomes: vesicular carriers for intercellular communication [J]. Curr Opin Cell Biol, 2009, 21(4):575-581. doi: 10.1016/j.ceb.2009.03.007
[12]	Simpson RJ. Exosomes: proteomic insights and diagnostic potential [J]. Expert Rev Proteomics, 2009, 6(3):267-283. doi: 10.1586/epr.09.17
[13]	Mitchell PJ, Welton J, Staffurth J. Can urinary exosomes act as treatment response markers in prostate cancer? [J]. J Transl Med, 2009, 7:4. doi: 10.1186/1479-5876-7-4 pmid: 19138409
[14]	Nilsson J, Skog J, Nordstrand A, et al. Prostate cancer-derived urine exosomes: a novel approach to biomarkers for prostate cancer [J]. Br J Cancer, 2009, 100(10):1603-1607. doi: 10.1038/sj.bjc.6605058
[15]	Théry C, Amigorena S, Raposo G, et al. Isolation and characte-rization of exosomes from cell culture supernatants and biological fluids [J]. Curr Protoc Cell Biol, 2006, Chapter 3: Unit 3.22.
[16]	Navabi H, Croston D, Hobot J, et al. Preperation of human ova-rian cancer ascites-derived exosomes for a clinical trial [J]. Blood Cells Mol Dis, 2005, 35(2):149-152. doi: 10.1016/j.bcmd.2005.06.008
[17]	Houali K, Wang X, Shimizu Y, et al. A new diagnostic marker for secreted Epstein-Barr virus encoded LMP1 and BARF1, oncoproteins in the serum and saliva of patients with nasopharyn-geal carcinoma [J]. Clin Cancer Res, 2007, 13(17):4993-5000. pmid: 17785549
[18]	Viaud S, Théry C, Ploix S, et al. Dendritic cell-derived exosomes for cancer immunotherapy: what’s next? [J]. Cancer Res, 2010, 70(4):1281-1285. doi: 10.1158/0008-5472.CAN-09-3276
[19]	Koga K, Matsumoto K, Akiyoshi T, et al. Purification, characte-rization and biological significance of tumor-derived exosomes [J]. Anticancer Res, 2005, 25(6A):3703-3707.
[20]	Zhou RL. LAPTM4B: a novel diagnostic biomarker and therapeutic target for hepatocellular carcinoma [M]// Wan-Yee Lau. Hepatocellular carcinoma: Basic research. Philippines: InTech Press, 2012: 1-34.
[21]	Yang Z, Senninger N, Flammang I, et al. Clinical impact of circulating LAPTM4B-35 in pancreatic ductal adenocarcinoma [J]. J Cancer Res Clin Oncol, 2019, 145(5):1165-1178. doi: 10.1007/s00432-019-02863-w

相关文章 15

[1]	丁婷婷,曾楚雄,胡丽娜,余明华. 基于癌症基因组图谱数据库结直肠癌免疫细胞浸润预测模型的建立[J]. 北京大学学报（医学版）, 2022, 54(2): 203-208.
[2]	王书磊,高阳旭,张宏武,杨海波,李辉,李宇,沈笠雪,姚红新. 儿童基底节区生殖细胞瘤30例临床分析[J]. 北京大学学报（医学版）, 2022, 54(2): 222-226.
[3]	林国中,马长城,吴超,司雨. 经锁孔入路显微外科切除腰椎管肿瘤54例临床研究[J]. 北京大学学报（医学版）, 2022, 54(2): 315-319.
[4]	李炳雨,唐祖南,胡耒豪,章文博,于尧,俞光岩,彭歆. 腮腺微小肿瘤的临床病理研究[J]. 北京大学学报（医学版）, 2022, 54(2): 335-339.
[5]	顾阳春,刘颖,谢超,曹宝山. 程序性死亡蛋白-1抑制剂治疗晚期肺癌出现垂体免疫不良反应3例[J]. 北京大学学报（医学版）, 2022, 54(2): 369-375.
[6]	李君,刘旭红,王工,程程,庄洪卿,杨瑞杰. 手臂位置对射波刀放射治疗脊柱肿瘤患者的剂量学影响[J]. 北京大学学报（医学版）, 2022, 54(1): 182-186.
[7]	杨朵,周心娜,王硕,王小利,袁艳华,杨化兵,耿会珍,彭兵,李子博,李彬,任军. 树突状细胞疫苗特异肿瘤多肽联合树突状细胞体外刺激淋巴细胞功能评估[J]. 北京大学学报（医学版）, 2021, 53(6): 1094-1098.
[8]	娄雪,廖莉,李兴珺,王楠,刘爽,崔若玫,徐健. 类风湿关节炎患者外周血TWEAK基因启动子区甲基化状态及其表达[J]. 北京大学学报（医学版）, 2021, 53(6): 1020-1025.
[9]	牛占岳,薛艳,张静,张贺军,丁士刚. 胃腺瘤性息肉的内镜和病理特点及癌变的危险因素分析[J]. 北京大学学报（医学版）, 2021, 53(6): 1122-1127.
[10]	叶剑飞,赵磊,王国良,洪锴,马潞林. 睾丸横纹肌肉瘤的诊治分析[J]. 北京大学学报（医学版）, 2021, 53(6): 1178-1182.
[11]	田雨,程晓悦,贺慧颖,王国良,马潞林. 肾细胞癌合并尿路瘤栓的临床病理特征: 6例报道及文献回顾[J]. 北京大学学报（医学版）, 2021, 53(5): 928-932.
[12]	孙争辉,黄晓娟,董靖晗,刘茁,颜野,刘承,马潞林. 临床T1期肾细胞癌肾窦侵犯的危险因素[J]. 北京大学学报（医学版）, 2021, 53(4): 659-664.
[13]	康文玉,王璐,邱敏,张帆,郭巍,强亚勇,拓鹏飞,宗有龙,刘磊磊,王帅帅. 肾上腺海绵状血管瘤1例及文献回顾[J]. 北京大学学报（医学版）, 2021, 53(4): 808-810.
[14]	林国中, 马长城, 王振宇, 谢京城, 刘彬, 陈晓东. 颈_1~2硬膜外神经鞘瘤的显微微创治疗[J]. 北京大学学报（医学版）, 2021, 53(3): 586-589.
[15]	杨榕,李庆祥,王逸飞,周闻,王雯,郭传瑸,刘浩,郭玉兴. 碘液浸染在Micro-CT下识别小鼠颅底-颞下区肿瘤组织中的应用[J]. 北京大学学报（医学版）, 2021, 53(3): 598-601.

Metrics

Viewed

Full text

Abstract

Cited

Shared

Discussed