Cell lines containing and expressing the human herpesvirus 6A ts gene are protected from both H-ras and BPV-1 transformation
Cell lines containing and expressing the human herpesvirus 6A ts gene are protected from both H-ras and BPV-1 transformation
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DOI：
10.1038/sj.onc.1200899
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发表时间：
1997-02-27
期刊：
影响因子：
8
通讯作者：
Rosenthal, LJ
中科院分区：
文献类型：
--
作者：
Araujo, JC;Doniger, J;Rosenthal, LJ
人疱疹病毒6A（HHV-6A）株U1102先前显示含有1473 bp的转化抑制基因（ts）（Araujo等人，1995），Ts在瞬时转染实验中抑制H-vas对NIH 3 T3细胞的转化以及H-uas和人免疫缺陷1型（HIV-1）启动子的转录。在本研究中，稳定表达ts蛋白的NIH 3 T3细胞系通过转染pRc-ts含有的TS基因的劳斯肉瘤病毒（RSV）的长末端重复序列（LTR）和新霉素选择标记的控制下，建立。选定的细胞系含有约1至2个拷贝/细胞的完整ts序列，表达ts蛋白，并以与亲本NIH 3 T3细胞大致相同的速率生长。这些细胞系被保护免于H-ras转化，而亲本和含有以反义方向克隆的ts基因的NIH 3 T3细胞则没有。氯霉素乙酰转移酶（CAT）基因的表达下的EJ-H-ras启动子的控制下，也被抑制在TS细胞系，但不是当CAT基因的小鼠骨肉瘤病毒LTR或人巨细胞病毒立即早期启动子的控制下。用逆转录病毒LNCts感染NIH 3 T3细胞，建立表达ts蛋白的细胞系，该细胞表达ts蛋白，并能保护细胞不受H-ras转化。此外，牛乳头瘤病毒1型（BPV-1）与ts共转染的细胞和转染BPV-1的表达ts的细胞系也能抑制BPV-1的转化。BPV-1 p89和p2443启动子在3 T3-ts系中下调。由于人乳头瘤病毒16型（HPV-16）p97启动子与BPV-1 p89启动子相似，因此还测试了ts抑制p97的能力。与H-ras和BPV-1启动子一样，HPV-16 p97在3 T3-ts系中下调。数据表明ts对H-ras、BPV-1和HPV-16启动子及其各自的癌基因的效用。
Human herpesvirus 6A (HHV-6A) strain U1102 was previously shown to contain a 1473 bp transformation suppressor gene (ts) (Araujo et al,, 1995), Ts inhibited transformation of NIH3T3 cells by H-vas and transcription of the H-uas and human immunodeficiency type 1 (HIV-1) promoters in transient transfection experiments. In the current study, stable NIH3T3 cell lines expressing ts protein were established by transfection with pRc-ts containing the ts gene under the control of the Rous sarcoma virus (RSV) long terminal repeat (LTR) and a neomycin selectable marker. Selected cell lines contained approximately one to two copies per cell of intact ts sequences, expressed ts protein and grew at approximately the same rate as parental NIH3T3 cells. These cell lines were protected from H-ras transformation while parental and NIH3T3 cells containing the ts gene cloned in the antisense orientation were not. Expression of the chloramphenicol acetyl transferase (CAT) gene under the control of the EJ-H-ras promoter was also suppressed in the ts cell lines but not when the CAT gene was under the control of the murine osteosarcoma virus LTR or human cytomegalovirus immediate early promoter. When NIH3T3 cell lines expressing ts protein were established by infection with the retrovirus, LNCts, the cells expressed ts protein and were protected from H-ras transformation, Furthermore, bovine papillomavirus type 1 (BPV-1) transformation was also suppressed in cells cotransfected with BPV-1 plus ts and in ts expressing cell lines transfected with BPV-1. The BPV-1 p89 and p2443 promoters were down-regulated in 3T3-ts lines. Because the human papillomavirus type 16 (HPV-16) p97 promoter has similarity to the BPV-1 p89 promoter, the ability of ts to suppress p97 was also tested. Like the H-ras and BPV-1 promoters, HPV-16 p97 was downregulated in 3T3-ts lines. The data indicate the utility of ts against H-ras, BPV-1 and HPV-16 promoters and their respective oncogenes.