ENHANCEMENT OF PULMONARY METASTASIS FORMATION AND GAMMA-GLUTAMYL-TRANSPEPTIDASE ACTIVITY IN B16 MELANOMA INDUCED BY DIFFERENTIATION INVITRO
ENHANCEMENT OF PULMONARY METASTASIS FORMATION AND GAMMA-GLUTAMYL-TRANSPEPTIDASE ACTIVITY IN B16 MELANOMA INDUCED BY DIFFERENTIATION INVITRO
复制标题
DOI：
10.1007/bf00121169
复制
发表时间：
1993-05-01
影响因子：
4
通讯作者：
GORELIK, E
中科院分区：
文献类型：
--
作者：
PREZIOSO, JA;WANG, N;GORELIK, E
与RPMI-1640培养基中的细胞相比，B16黑素瘤亚系（B16-F10-BL 6和B16-F1）在Dulbecco改良Eagle培养基（DMEM）中培养时表现出升高的腺苷3 '，5'-环一磷酸（cAMP）水平。与此同时，在DMEM中培养的细胞具有增加的酪氨酸酶活性、黑化和树突形成，这些都是黑素瘤分化的标志物。此外，当在DMEM中培养时，两种细胞系的增殖率相对于维持在RPMI-1640培养基中的细胞降低80-85%。在这些研究中，在DMEM中发现的黑色素前体酪氨酸（Tyr）和苯丙氨酸（Phe）的水平升高被证明不是用DMEM观察到的表型变化的唯一原因。BL 6和B16-F1细胞系在同系C57 BL/6小鼠中培养时，在DMEM培养基中比在RPMI-1640培养基中形成更多的实验性肺肿瘤转移。在具有正常和耗尽的自然杀伤（NK）细胞活性的裸鼠中的转移形成的分析显示，在DMEM中维持的BL 6细胞的增强的肺定殖能力独立于T细胞或NK细胞介导的免疫的功能。观察到测试品系的转移能力与膜相关酶γ-谷氨酰转肽酶（γ-GTdR，EC 2.3.2.2）的表达之间的密切关联。高转移性BL 6细胞系的γ-GT α活性水平比弱转移性B16-F1细胞系高20倍。当在DMEM中生长时，两种细胞系都具有升高的γ-GT α活性，这增加了转移能力。与在RPMI-1640培养基中生长的细胞相比，在DMEM中维持的变体B16黑色素瘤细胞的肺定殖能力的显著变化可以作为理解触发细胞分化以及转移发展所涉及的某些步骤的有用模型。
B16 melanoma sublines (B16-F10-BL6 and B16-F1) exhibited elevated adenosine 3',5'-cyclic monophosphate (cAMP) levels when cultured in Dulbecco's modified Eagle's medium (DMEM) in comparison to cells in RPMI-1640 medium. In parallel, cells cultured in DMEM had increased tyrosinase activity, melanization and dendrite formation, all markers of melanoma differentiation. Also, the proliferative rates of both cell lines were decreased by 80-85% when cultured in DMEM relative to cells maintained in RPMI-1640 medium. In these studies, elevated levels of the melanin precursors tyrosine (Tyr) and phenylalanine (Phe) found in DMEM were shown not to be solely responsible for the phenotypic changes observed with DMEM. Both BL6 and B16-F1 cell lines formed more experimental pulmonary tumor metastasis in syngeneic C57BL/6 mice when maintained in DMEM vs RPMI-1640 medium. Analysis of metastasis formation in nude mice with normal and depleted natural killer (NK) cell activity revealed that the enhanced lung colonizing capacity of the BL6 cells maintained in DMEM was independent of the function of T-cell or NK-cell-mediated immunity. A close association between metastatic ability of tested lines and the expression of the membrane-associated enzyme gamma-glutamyltranspeptidase (gamma-GTPase, EC 2.3.2.2) was observed. The highly metastatic BL6 cell line had 20-fold higher levels of gamma-GTPase activity than the weakly metastatic B16-F1 cell line. Both cell lines, when grown in DMEM, had elevated gamma-GTPase activity that paralleled augmentation of metastatic ability. The dramatic changes in lung-colonizing capacity of the variant B16 melanoma celts maintained in DMEM in contrast to those grown in RPMI-1640 medium may serve as a useful model in understanding certain steps involved in triggering cell differentiation as well as metastasis development.