Transfection with the inducible nitric oxide synthase gene suppresses tumorigenicity and abrogates metastasis by K-1735 murine melanoma cells.
Transfection with the inducible nitric oxide synthase gene suppresses tumorigenicity and abrogates metastasis by K-1735 murine melanoma cells.
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诱导型一氧化氮合酶基因转染可抑制 K-1735 鼠黑色素瘤细胞的致瘤性并消除转移。
DOI：
10.1084/jem.181.4.1333
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发表时间：
1995-04-01
影响因子：
15.3
通讯作者：
Fidler, Isaiah J.
中科院分区：
文献类型：
--
作者：
Xie, Keping;Huang, Suyun;Dong, Zhongyun;Juang, Shin-Hun;Gutman, Mordechai;Xie, Qiao-Wen;Nathan, Carl;Fidler, Isaiah J.
我们实验室以前的研究表明，诱导型一氧化氮合酶（iNOS）的表达水平和小鼠K-1735黑色素瘤细胞的转移潜力之间呈反比关系。本研究的目的是提供直接的证据表明，iNOS的表达抑制黑色素瘤细胞的转移潜力。在含有3 mM NG-甲基-L-精氨酸（NMA）的培养基中，用功能性iNOS（C4.L8）、失活突变的iNOS（C4.S2）或新霉素抗性（C4.Neo）基因转染表达低水平iNOS的高转移性K-1735克隆4细胞（C4.P）。通过Southern和北方印迹分析以及用特异性抗iNOS单克隆抗体的均匀染色鉴定阳性转染子。收获C4.P（亲本）、C4.Neo.3（对照转染）、C4.S2.3（失活iNOS）和C4.L8.5（功能性iNOS）的半流培养物，并将活细胞静脉内注射到同基因C3 H/HeN小鼠和同种异体BALB/c裸鼠中。C4.P、C4.Neo.3和C4.S2.3细胞具有高度转移性，而C4.L8.5细胞无转移性。用[125 I] dUrd标记的肿瘤细胞进行的实验表明，肺微血管系统中的初始停滞在各细胞系之间没有差异，但C4.L8.5细胞在注射后48-72 h死亡。在每天两次注射20 mg NMA的小鼠中发现所有K-1735 C4细胞（包括C4.L8.5）的存活率增加。C4.L8.5细胞在裸鼠中产生缓慢生长的皮下肿瘤，而其他三个细胞系产生快速生长的肿瘤。体外研究证实，在没有NMA的情况下，C4.L8.5细胞中iNOS的表达诱导凋亡。总的来说，这些数据表明，在黑色素瘤细胞中的重组iNOS的表达与细胞凋亡，抑制肿瘤发生，并废除转移。
Previous studies from our laboratory demonstrated an inverse relationship between the expression level of inducible nitric oxide synthase (iNOS) and the metastatic potential of murine K-1735 melanoma cells. The purpose of this study was to provide direct evidence that the expression of iNOS suppresses metastatic potential of melanoma cells. Highly metastatic K-1735 clone 4 cells (C4.P), which express low levels of iNOS, were transfected with a functional iNOS (C4.L8), inactive-mutated iNOS (C4.S2), or neomycin-resistance (C4.Neo) genes in medium containing 3 mM NG-methyl-L-arginine (NMA). Positive transfectants were identified by Southern and Northern blot analyses and homogeneous staining with a specific anti-iNOS monoclonal antibody. Semiconfluent cultures of C4.P (parental), C4.Neo.3 (control transfection), C4.S2.3 (inactive iNOS), and C4.L8.5 (functional iNOS) were harvested, and viable cells were injected intravenously into syngeneic C3H/HeN mice and allogeneic BALB/c nude mice. C4.P, C4.Neo.3, and C4.S2.3 cells were highly metastatic whereas C4.L8.5 cells were not metastatic. Experiments with [125I]dUrd-labeled tumor cells demonstrated that the initial arrest in the lung microvasculature did not differ among the lines, but that C4.L8.5 cells died by 48-72 h after injection. Enhanced survival of all K-1735 C4 cells (including C4.L8.5) was found in mice given twice daily injections of 20 mg NMA. The C4.L8.5 cells produced slow growing subcutaneous tumors in nude mice, whereas the other three lines produced fast growing tumors. In vitro studies confirmed that in the absence of NMA the expression of iNOS in C4.L8.5 cells induced apoptosis. Collectively, these data demonstrate that the expression of recombinant iNOS in melanoma cells is associated with apoptosis, suppression of tumorigenicity, and abrogation of metastasis.
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影响因子：
56.9
作者：
KARUPIAH, G;XIE, QW;MACMICKING, JD
通讯作者：
MACMICKING, JD
影响因子：
64.8
作者：
PALMER, RMJ;ASHTON, DS;MONCADA, S
通讯作者：
MONCADA, S
影响因子：
56.9
作者：
HIBBS, JB;TAINTOR, RR;VAVRIN, Z
通讯作者：
VAVRIN, Z
DOI：
10.1073/pnas.88.19.8485
发表时间：
1991-10-01
影响因子：
11.1
作者：
HOGQUIST, KA;NETT, MA;CHAPLIN, DD
通讯作者：
CHAPLIN, DD
DOI：
10.1073/pnas.84.24.9265
发表时间：
1987-12-01
影响因子：
11.1
作者：
IGNARRO, LJ;BUGA, GM;CHAUDHURI, G
通讯作者：
CHAUDHURI, G