MAN2A1-FER Fusion Gene Is Expressed by Human Liver and Other Tumor Types and Has Oncogenic Activity in Mice.
MAN2A1-FER Fusion Gene Is Expressed by Human Liver and Other Tumor Types and Has Oncogenic Activity in Mice.
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MAN2A1-FER融合基因由人肝脏和其他肿瘤类型表达，并且在小鼠中具有致癌活性。
DOI：
10.1053/j.gastro.2016.12.036
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发表时间：
2017-10
期刊：
影响因子：
29.4
通讯作者：
Luo JH
中科院分区：
文献类型：
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作者：
Chen ZH;Yu YP;Tao J;Liu S;Tseng G;Nalesnik M;Hamilton R;Bhargava R;Nelson JB;Pennathur A;Monga SP;Luketich JD;Michalopoulos GK;Luo JH
人类肿瘤和肝癌细胞系表达甘露糖苷酶α 2A类成员1基因（MAN 2A 1）中的前13个外显子与FER酪氨酸激酶基因（FER）中的后6个外显子之间的融合产物，称为MAN 2A 1-FER。我们研究了MAN 2A 1-FER是否在人类肝肿瘤中表达及其在肝癌发生中的作用。我们对匹兹堡大学收集的102例非小细胞肺肿瘤、61例卵巢肿瘤、70例肝肿瘤、156例多形性胶质母细胞瘤样本、27例食管腺癌和269例前列腺癌样本以及10例非肿瘤肝组织和20例非肿瘤前列腺组织进行了逆转录PCR分析。我们还测量了15种人类癌细胞系的表达。我们在NIH 3 T3和HEP 3B（肝癌）细胞中表达了标记形式的MAN 2A 1-FER;分离高尔基体进行分析。MAN 2A 1-FER也在PC 3或DU 145（前列腺癌）、NIH 3 T3（成纤维细胞）、H23（肺癌）和A-172（多形性胶质母细胞瘤）细胞系中过表达，并在HUH 7（肝癌）细胞中敲除。分析细胞的增殖和侵袭测定，和/或注射到SCID小鼠的侧腹;评估异种移植肿瘤生长和转移。对肝缺失PTEN的小鼠给予尾静脉注射MAN 2A 1-FER。我们检测到MAN 2A 1-FER mRNA和融合蛋白（114 kD）在肝细胞癌细胞系HUH 7，以及在肝肿瘤，食管腺癌，多形性胶质母细胞瘤，前列腺肿瘤，非小细胞肺肿瘤，卵巢肿瘤，但不是非肿瘤前列腺或肝组织。MAN 2A 1-FER蛋白保留了MAN 2A 1的高尔基体定位信号肽，并在癌细胞系中从细胞质易位到高尔基体。MAN 2A 1-FER的酪氨酸激酶活性几乎是野生型FER的4倍，并且在其N-末端的酪氨酸88处磷酸化表皮生长因子受体（EGFR）。MAN 2A 1-FER在4种细胞系中的表达导致BRAF、MEK和AKT的EGFR活化; MAN 2A 1-FER敲除的HUH 7细胞中这些蛋白的磷酸化显著降低。与不表达融合蛋白的细胞相比，表达MAN 2A 1-FER的细胞系的增殖、集落形成和侵袭力增加，并在小鼠中形成更大（2倍以上）的异种移植肿瘤，转移灶更多。与对照HUH 7细胞相比，MAN 2A 1-FER敲除的HUH 7细胞形成更小的异种移植肿瘤，具有更少的转移。表达MAN 2A 1-FER的HUH 7、A-172和PC 3细胞对FER激酶抑制剂克唑替尼和EGFR激酶抑制剂canertinib的敏感性约为2倍;这些药物减缓了MAN 2A 1-FER细胞的异种移植肿瘤的生长，并阻止了它们在小鼠中的转移。水动力尾静脉注射MAN 2A 1-FER导致Pten肝破坏的小鼠中肝癌的快速发展。许多人类肿瘤类型和癌细胞系表达MAN 2A 1-FER融合体，其增加癌细胞系的增殖和侵袭力，并且在小鼠中具有肝致癌活性。
Human tumors and liver cancer cell lines express the product of a fusion between the first 13 exons in the mannosidase alpha class 2A member 1 gene (MAN2A1) and the last 6 exons in the FER tyrosine kinase gene (FER), called MAN2A1–FER. We investigated whether MAN2A1–FER is expressed by human liver tumors and its role in liver carcinogenesis. We performed reverse transcription PCR analyses of 102 non-small cell lung tumors, 61 ovarian tumors, 70 liver tumors, 156 glioblastoma multiform samples, 27 esophageal adenocarcinomas, and 269 prostate cancer samples, as well as 10 non-tumor liver tissues and 20 non-tumor prostate tissues, collected at the University of Pittsburgh. We also measured expression by 15 human cancer cell lines. We expressed a tagged form of MAN2A1–FER in NIH3T3 and HEP3B (liver cancer) cells; Golgi were isolated for analysis. MAN2A1–FER was also overexpressed in PC3 or DU145 (prostate cancer), NIH3T3 (fibroblast), H23 (lung cancer) and A-172 (glioblastoma multiforme) cell lines and knocked out in HUH7 (liver cancer) cells. Cells were analyzed for proliferation and in invasion assays, and/or injected into flanks of SCID mice; xenograft tumor growth and metastasis were assessed. Mice with hepatic deletion of PTEN were given tail-vein injections of MAN2A1–FER. We detected MAN2A1–FER mRNA and fusion protein (114 kD) in the hepatocellular carcinoma cell line HUH7, as well as in liver tumors, esophageal adenocarcinoma, glioblastoma multiforme, prostate tumors, non-small cell lung tumors, and ovarian tumors, but not non-tumor prostate or liver tissues. MAN2A1–FER protein retained the signal peptide for Golgi localization from MAN2A1 and translocated from the cytoplasm to Golgi in cancer cell lines. MAN2A1–FER had tyrosine kinase activity almost 4-fold higher than that of wild-type FER, and phosphorylated the epidermal growth factor receptor (EGFR) at tyrosine 88 in its N-terminus. Expression of MAN2A1–FER in 4 cell lines led to EGFR activation of BRAF, MEK, and AKT; HUH7 cells with MAN2A1–FER knockout had significant decreases in phosphorylation of these proteins. Cell lines that expressed MAN2A1–FER had increased proliferation, colony formation, and invasiveness and formed larger (more than 2-fold) xenograft tumors in mice, with more metastases, than cells not expressing the fusion protein. HUH7 cells with MAN2A1–FER knockout formed smaller xenograft tumors, with fewer metastases, than control HUH7 cells. HUH7, A-172, and PC3 cells that expressed MAN2A1–FER were about 2-fold more sensitive to the FER kinase inhibitor crizotinib and the EGFR kinase inhibitor canertinib; these drugs slowed growth of xenograft tumors from MAN2A1–FER cells and prevented their metastasis in mice. Hydrodynamic tail-vein injection of MAN2A1–FER resulted in rapid development of liver cancer in mice with hepatic disruption of Pten. Many human tumor types and cancer cell lines express the MAN2A1–FER fusion, which increases proliferation and invasiveness of cancer cell lines and has liver oncogenic activity in mice.
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影响因子：
8
作者：
Ren, B;Yu, G;Luo, JH
通讯作者：
Luo, JH
影响因子：
4.8
作者：
Chen, Zhang-Hui;Yu, Yan P.;Luo, Jian-Hua
通讯作者：
Luo, Jian-Hua
DOI：
10.1073/pnas.92.25.11766
发表时间：
1995-12-05
影响因子：
11.1
作者：
MISAGO, M;LIAO, YF;FUKUDA, MN
通讯作者：
FUKUDA, MN
影响因子：
7.3
作者：
Han, Yu-Chen;Zheng, Zhong-Liang;Zuo, Ze-Hua;Yu, Yan P.;Chen, Rui;Tseng, George C.;Nelson, Joel B.;Luo, Jian-Hua
通讯作者：
Luo, Jian-Hua
影响因子：
6.4
作者：
通讯作者：
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