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首页> 外文期刊>The Journal of Steroid Biochemistry and Molecular Biology >Identification of twenty alternatively spliced estrogen receptor alpha mRNAs in breast cancer cell lines and tumors using splice targeted primer approach.
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Identification of twenty alternatively spliced estrogen receptor alpha mRNAs in breast cancer cell lines and tumors using splice targeted primer approach.

机译：使用剪接靶向引物方法鉴定乳腺癌细胞系和肿瘤中的二十种选择性剪接的雌激素受体αmRNA。

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摘要

Estrogen receptor (ER) alpha splice variant transcript profiles were analyzed by RT PCR in six ER positive breast cancer cell lines, MCF-7, T47D, ZR-75, LCC1, LCC2 and LCC9, three ER negative cell lines, MDA-MB-435, MDA-MB-235 and LCC6, and three ER positive malignant breast tumors using targeted primers which specifically anneal to the splice junctions of exon 2Delta, exon 3Delta, exons 2-3Delta, exon 4Delta, exon 5Delta, exon 6Delta and exon 7Delta. The partner primers were chosen such that largest possible transcripts were amplified between exons 1 and 8. The results described here show that each splice specific primer amplified not only the single exon deleted transcript but also a number of related transcripts that have deletions in various combinations of exons. The exon 2Delta specific primer amplified five transcripts that have deletions in exon 2, exons 2 and 7, exons 2, 5, and 7, exons 2 and 4-5, and exons 2 and 4-6. The exon 3Delta specific primer amplified two transcripts that have deletions in exon 3, and exons 3 and 7. The exon 2-3Delta specific primer amplified three products that have deletions in exons 2-3, exons 2-3 and 7 and exons 2-3, 5 and 7. The exon 4Delta specific primer amplified two products that have deletions in exon 4, and exons 4 and 7. The exon 5Delta specific primer amplified three transcripts, that have deletions in exon 5, exons 5 and 2, and exons 5, and 2-3. The 6Delta specific primer amplified only one transcript that has a deletion in exon 6. The 7Delta specific primer amplified four transcripts, that have deletions in exon 7, exons 7 and 4, exons 7 and 3-4, and exons 7 and 3-5. None of the above splice specific primers amplified the wild type ER sequences. The six ER positive cell lines differed in the patterns of the variant transcripts and among the three ER negative cell lines analyzed, only MDA-MB-435 showed the presence of exon 2Delta and exon 4Delta transcripts. Analyses in the tumor samples indicated that the above transcripts are extensively modified.
机译：通过RT PCR在6种ER阳性乳腺癌细胞系MCF-7，T47D，ZR-75，LCC1，LCC2和LCC9、3种ER阴性细胞系，MDA-MB-中对雌激素受体（ER）α剪接变体转录谱进行了分析。 435，MDA-MB-235和LCC6以及三种ER阳性恶性乳腺肿瘤，使用靶向引物进行特异性退火，该引物与外显子2Delta，外显子3Delta，外显子2-3Delta，外显子4Delta，外显子5Delta，外显子6Delta和外显子7Delta的剪接点退火。选择伴侣引物以使最大可能的转录物在外显子1和8之间扩增。此处描述的结果表明，每个剪接特异性引物不仅扩增单个外显子缺失的转录物，而且扩增了许多相关转录物，这些转录物具有多种组合形式的缺失。外显子。外显子2Delta特异性引物扩增了在外显子2，外显子2和7，外显子2、5和7，外显子2和4-5以及外显子2和4-6具有缺失的五个转录物。外显子3Delta特异性引物扩增了两个在外显子3以及外显子3和7缺失的转录物。外显子2-3Delta特异性引物扩增了在外显子2-3，外显子2-3和7以及外显子2-缺失的三种产物。 3、5和7。外显子4Delta特异性引物扩增了两个在外显子4和7外显子上都缺失的产物。外显子5Delta特异性引物扩增了三个在外显子5，外显子5和2，外显子上有缺失的转录本。 5和2-3。 6Delta特异性引物仅扩增了一个外显子6缺失的转录物。7Delta特异性引物扩增了四个在外显子7，外显子7和4，外显子7和3-4，外显子7和3-5缺失的转录物。 。上述剪接特异性引物均未扩增野生型ER序列。六个ER阳性细胞系的变异转录本模式有所不同，在所分析的三个ER阴性细胞系中，只有MDA-MB-435显示存在外显子2Delta和外显子4Delta转录本。肿瘤样品中的分析表明上述转录物被广泛修饰。

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