Fasco M J
Wadsworth Center, New York State Department of Health, Albany 12201-0509, USA.
Mol Cell Endocrinol. 1998 Mar 16;138(1-2):51-9. doi: 10.1016/s0303-7207(98)00048-3.
Relative proportions of the estrogen receptor (ER) alternatively spliced mRNA variants from the proximal (A) and distal (B) promoter pre-mRNA transcripts were measured in normal human uterus, an endometrial tumor, and in T47D, MCF-7, and BT-20 breast tumor cell lines. A single tube RNA-PCR method was developed to determine the proportions of the individual transcripts and a nested, competitive RNA-PCR method to determine the proportions of the alternatively spliced variants. Except for the BT-20 cells, the patterns of splice variants produced from each transcript were very similar. In BT-20 cells no splice variants were detected for the minor (< or = 1%) A promoter transcript, although the B promoter transcript was alternatively spliced similarly to the other samples, with the exon 7 variant as the major mRNA form. These results indicate that the mRNA spliced variant patterns in most tissues and tumors will be essentially unaffected by any changes in the A and B promoter ER mRNA transcript ratios that may occur. At least one exception does exist, however, and only more comprehensive studies can determine whether the BT-20 cells are unique or part of a larger subgroup.
在正常人体子宫、一种子宫内膜肿瘤以及T47D、MCF - 7和BT - 20乳腺癌细胞系中,测量了来自近端(A)和远端(B)启动子前体mRNA转录本的雌激素受体(ER)可变剪接mRNA变体的相对比例。开发了一种单管RNA - PCR方法来确定各个转录本的比例,并开发了一种巢式竞争性RNA - PCR方法来确定可变剪接变体的比例。除了BT - 20细胞外,每个转录本产生的剪接变体模式非常相似。在BT - 20细胞中,未检测到少量(≤1%)A启动子转录本的剪接变体,尽管B启动子转录本的可变剪接与其他样品相似,外显子7变体是主要的mRNA形式。这些结果表明,大多数组织和肿瘤中的mRNA剪接变体模式基本上不会受到A和B启动子ER mRNA转录本比例可能发生的任何变化的影响。然而,至少存在一个例外,只有更全面的研究才能确定BT - 20细胞是独特的还是更大亚组的一部分。