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在转染了I类主要组织相容性复合体基因的BL6黑色素瘤细胞中,脑池内A型逆转录病毒颗粒的丢失。

Loss of intracisternal A-type retroviral particles in BL6 melanoma cells transfected with MHC class I genes.

作者信息

Li M, Muller J, Rao V, Hearing V, Lueders K, Gorelik E

机构信息

Pittsburgh Cancer Institute and Department of Pathology, University of Pittsburgh, PA 15213, USA.

出版信息

J Gen Virol. 1996 Nov;77 ( Pt 11):2757-65. doi: 10.1099/0022-1317-77-11-2757.

Abstract

Electron microscopy of B16 melanoma and its sublines revealed that these cells produce numerous intracisternal A-type retroviral particles (IAPs). To identify and sequence the melanoma-associated IAPs of C57BL/6 mice, a cDNA library was constructed from IAP-producing BL6.8 cell RNA and screened using MIA14 IAP DNA as a probe. A 6-8 kb mRNA was identified that represents the full-length message for a new subfamily of IAP, termed MeIAP. The melanoma-derived IAP cDNA showed high similarity to MIA14 with major differences in the LTR. A nine base motif of the R region showed that this IAP differs from other previously sequenced IAPs. Analysis of the individual clones from BL6 melanoma revealed that IAPs were produced only in the clones that failed to express H-2Kb molecules. No IAPs were found in the melanoma clones that expressed endogenous H-2Kb. To analyse further the association between MHC class I genes and IAP production, the H-2Kb-negative clones of BL6 melanoma were transfected with various H-2 genes. Transfection of the H-2Kb or H-2Kd, but not H-2Dd or H-2Ld genes resulted in the elimination of IAPs. Northern blot analysis revealed that loss of IAPs in the H-2K gene-transfected BL6 melanoma cells was due to lack of IAP transcripts. Elimination of IAPs in the H-2Kb-positive BL6 melanoma cells was also accompanied by alterations in expression of various cellular genes and changes of their phenotypic properties.

摘要

对B16黑色素瘤及其亚系进行电子显微镜检查发现,这些细胞产生大量A型核内逆转录病毒颗粒(IAP)。为了鉴定和测序C57BL/6小鼠的黑色素瘤相关IAP,从产生IAP的BL6.8细胞RNA构建了一个cDNA文库,并使用MIA14 IAP DNA作为探针进行筛选。鉴定出一个6 - 8 kb的mRNA,它代表了一个新的IAP亚家族的全长信息,称为MeIAP。黑色素瘤来源的IAP cDNA与MIA14高度相似,但在长末端重复序列(LTR)上有主要差异。R区域的一个九碱基基序表明,这种IAP与其他先前测序的IAP不同。对BL6黑色素瘤的单个克隆进行分析发现,IAP仅在不表达H - 2Kb分子的克隆中产生。在表达内源性H - 2Kb的黑色素瘤克隆中未发现IAP。为了进一步分析MHC I类基因与IAP产生之间的关联,用各种H - 2基因转染BL6黑色素瘤的H - 2Kb阴性克隆。转染H - 2Kb或H - 2Kd基因,但不是H - 2Dd或H - 2Ld基因,导致IAP消除。Northern印迹分析表明,H - 2K基因转染的BL6黑色素瘤细胞中IAP的缺失是由于缺乏IAP转录本。H - 2Kb阳性的BL6黑色素瘤细胞中IAP的消除还伴随着各种细胞基因表达的改变及其表型特性的变化。

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