Domenzain Clelia, Docampo María José, Serra Montserrat, Miquel Laia, Bassols Anna
Departament de Bioqui;mica i Biologia Molecular, Facultat de Veterinària, Universitat Autònoma de Barcelona, Bellaterra 081893, Spain
Biochim Biophys Acta. 2003 Sep 23;1642(1-2):107-14. doi: 10.1016/s0167-4889(03)00104-6.
Versican is a large chondroitin sulfate proteoglycan produced by human melanoma cell lines and malignant melanocytic lesions. In the present work, we have analyzed the expression of versican spliced variants V0, V1, V2 and V3 in human melanoma cell lines at several differentiation degrees. The isoform expression pattern depends on the degree of cell differentiation. Differentiated cell lines do not produce any of the versican isoforms as analyzed by Western blot, Northern blot and RT-PCR. All cell lines with an early or intermediate degree of differentiation (AX3, SK-mel-37, Rider, SK-mel-1.36-1-5 and SK-mel-3.44) expressed V0 and V1 transcripts, whereas V2 and V3 expression was shown only by the undifferentiated cell lines SK-mel-1.36-1-5 and Rider. Furthermore, we have analyzed the expression of versican isoforms in SK-mel-3.44 and SK-mel-1.36-1-5 cells induced to differentiate by TPA treatment. The expression of the large V0, V1 and V2 isoforms practically disappears in differentiated cells, whereas V3 remains detectable by RT-PCR analysis.
多功能蛋白聚糖是一种由人黑色素瘤细胞系和恶性黑素细胞病变产生的大型硫酸软骨素蛋白聚糖。在本研究中,我们分析了多功能蛋白聚糖剪接变体V0、V1、V2和V3在不同分化程度的人黑色素瘤细胞系中的表达情况。异构体表达模式取决于细胞分化程度。通过蛋白质印迹法、Northern印迹法和逆转录聚合酶链反应分析,分化的细胞系不产生任何多功能蛋白聚糖异构体。所有早期或中期分化程度的细胞系(AX3、SK-mel-37、Rider、SK-mel-1.36-1-5和SK-mel-3.44)均表达V0和V1转录本,而只有未分化的细胞系SK-mel-1.36-1-5和Rider显示出V2和V3表达。此外,我们分析了经佛波酯处理诱导分化的SK-mel-3.44和SK-mel-1.36-1-5细胞中多功能蛋白聚糖异构体的表达。在分化细胞中,大型V0、V1和V2异构体的表达几乎消失,而通过逆转录聚合酶链反应分析仍可检测到V3。