Choline Kinase α Mediates Interactions Between the Epidermal Growth Factor Receptor and Mechanistic Target of Rapamycin Complex 2 in Hepatocellular Carcinoma Cells to Promote Drug Resistance and Xenograft Tumor Progression.
Choline Kinase α Mediates Interactions Between the Epidermal Growth Factor Receptor and Mechanistic Target of Rapamycin Complex 2 in Hepatocellular Carcinoma Cells to Promote Drug Resistance and Xenograft Tumor Progression.
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胆碱激酶 α 介导肝细胞癌细胞中表皮生长因子受体与雷帕霉素复合物 2 机制靶点之间的相互作用，促进耐药性和异种移植肿瘤进展
DOI：
10.1053/j.gastro.2016.12.033
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发表时间：
2017-04
期刊：
影响因子：
29.4
通讯作者：
Chen L
中科院分区：
文献类型：
--
作者：
Lin XM;Hu L;Gu J;Wang RY;Li L;Tang J;Zhang BH;Yan XZ;Zhu YJ;Hu CL;Zhou WP;Li S;Liu JF;Gonzalez FJ;Wu MC;Wang HY;Chen L
胆碱激酶α（CHKA）催化胆碱转化为磷酸胆碱，并可促进致癌作用。CHKA在肝细胞癌（HCC）发病机制中的作用知之甚少。我们对9对HCC和非肿瘤相邻组织进行了全外显子组和转录组序列分析。我们对2006年1月至2009年12月在中国上海接受手术的120例原发性肝癌和非肿瘤相邻组织进行了组织芯片分析;还分析了48组标本（肝癌和非肿瘤相邻组织）。CHKA基因拷贝数进行定量，并通过定量逆转录聚合酶链反应分析验证结果。CHKA信使RNA和蛋白质水平通过聚合酶链反应、荧光化学和免疫印迹分析测定。CHKA在2个肝细胞系和7个肝癌细胞系中进行了检测，并在3个肝癌细胞系中用小干扰RNA敲低。在增殖、伤口愈合、迁移和侵袭测定中分析细胞。将细胞注射到小鼠的尾静脉中，并定量肿瘤生长和转移。进行免疫沉淀和免疫荧光测定以确定CHKA与表皮生长因子受体（EGFR）和雷帕霉素复合物2的机制靶标之间的相互作用。与非肿瘤组织相比，CHKA信使RNA的水平在HCC组织中经常增加;增加的表达与CHKA位点的扩增相关。表达高水平CHKA的肿瘤具有更具侵袭性的表型，与肿瘤表达低水平CHKA的患者相比，患有这些肿瘤的患者术后生存时间较短。稳定过表达CHKA的HCC细胞系比对照HCC细胞具有更高水平的迁移和侵袭，并且与不过表达CHKA的HCC细胞相比，在小鼠中形成具有更多转移的更大的异种移植肿瘤。CHKA是EGFR与雷帕霉素复合物2的机制靶点之间的物理相互作用所必需的。这种复合物是HCC细胞在小鼠中形成转移性异种移植肿瘤和对EGFR抑制剂产生耐药性所必需的。我们发现，与非肿瘤组织相比，CHKA在人类HCC中的水平增加，并且增加的表达与肿瘤侵袭性和患者生存时间缩短相关。CHKA在HCC细胞系中的过表达通过促进EGFR与雷帕霉素复合物2的机制靶点的相互作用而增加了它们的侵袭性、对EGFR抑制剂的抗性以及在小鼠中形成转移性肿瘤的能力。
Choline kinase α (CHKA) catalyzes conversion of choline to phosphocholine and can contribute to carcinogenesis. Little is known about the role of CHKA in the pathogenesis of hepatocellular carcinoma (HCC). We performed whole-exome and transcriptome sequence analyses of 9 paired HCC and non—tumor-adjacent tissues. We performed tissue chip analyses of 120 primary HCC and non—tumor-adjacent tissues from patients who received surgery in Shanghai, China from January 2006 through December 2009; 48 sets specimens (HCC and non—tumor-adjacent tissues) were also analyzed. CHKA gene copy number was quantified and findings were validated by quantitative reverse transcription polymerase chain reaction analysis. CHKA messenger RNA and protein levels were determined by polymerase chain reaction, immunohisto-chemical, and immunoblot analyses. CHKA was examined in 2 hepatocyte cell lines and 7 HCC-derived cell lines, and knocked down with small interfering RNAs in 3 HCC cell lines. Cells were analyzed in proliferation, wound healing, migration, and invasion assays. Cells were injected into tail veins of mice and tumor growth and metastasis were quantified. Immunoprecipitation and immunofluorescence assays were conducted to determine interactions between CHKA and the epidermal growth factor receptor (EGFR) and the mechanistic target of rapamycin complex 2. Levels of CHKA messenger RNA were frequently increased in HCC tissues compared with nontumtissues; increased expression was associated with amplification at the CHKA loci. Tumors that expressed high levels of CHKA had more aggressive phenotypes, and patients with these tumors had shorter survival times after surgery compared to patients whose tumors expressed low levels of CHKA. HCC cell lines that stably overexpressed CHKA had higher levels of migration and invasion than control HCC cells, and formed larger xenograft tumors with more metastases in mice compared to HCC cells that did not overexpress CHKA. CHKA was required for physical interaction between EGFR and mechanistic target of rapamycin complex 2. This complex was required for HCC cells to form metastatic xenograft tumors in mice and to become resistant to EGFR inhibitors. We found levels of CHKA to be increased in human HCCs compared to nontumor tissues, and increased expression to be associated with tumor aggressiveness and reduced survival times of patients. Overexpression of CHKA in HCC cell lines increased their invasiveness, resistance to EGFR inhibitors, and ability to form metastatic tumors in mice by promoting interaction of EGFR with mechanistic target of rapamycin complex 2.
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影响因子：
8
作者：
Hernando, E.;Sarmentero-Estrada, J.;Koppie, T.;Belda-Iniesta, C.;de Molina, V. Ramirez;Cejas, P.;Ozu, C.;Le, C.;Sanchez, J. J.;Gonzalez-Baron, M.;Koutcher, J.;Cordon-Cardo, C.;Bochner, B. H.;Lacal, J. C.;de Molina, A. Ramirez
通讯作者：
de Molina, A. Ramirez
影响因子：
37.3
作者：
Chua BT;Gallego-Ortega D;Ramirez de Molina A;Ullrich A;Lacal JC;Downward J
通讯作者：
Downward J
DOI：
10.1016/j.bbalip.2014.01.007
发表时间：
2014-04-01
影响因子：
4.8
作者：
Domizi, Pablo;Aoyama, Chieko;Banchio, Claudia
通讯作者：
Banchio, Claudia
DOI：
10.1016/s0006-291x(02)00920-8
发表时间：
2002-08-23
影响因子：
3.1
作者：
Ramírez de Molina, A;Rodríguez-González, A;Lacal, JC
通讯作者：
Lacal, JC
影响因子：
51.1
作者：
Ramirez de Molina, Ana;Sormentero-Estrada, Jacinto;Lacal, Juan Carlos
通讯作者：
Lacal, Juan Carlos