Dube Syamalima, Yalamanchili Santhi, Lachant Joseph, Abbott Lynn, Benz Patricia, Mitschow Charles, Dube Dipak K, Poiesz Bernard J
Division of Hematology/Oncology, Department of Medicine, SUNY Upstate Medical University, Syracuse, NY 13210, USA.
Int J Breast Cancer. 2015;2015:859427. doi: 10.1155/2015/859427. Epub 2015 Jun 11.
Nine malignant breast epithelial cell lines and 3 normal breast cell lines were examined for stress fiber formation and expression of TPM1 isoform-specific RNAs and proteins. Stress fiber formation was strong (++++) in the normal cell lines and varied among the malignant cell lines (negative to +++). Although TPM1γ and TPM1δ were the dominant transcripts of TPM1, there was no clear evidence for TPM1δ protein expression. Four novel human TPM1 gene RNA isoforms were discovered (λ, μ, ν, and ξ), which were not identified in adult and fetal human cardiac tissues. TPM1λ was the most frequent isoform expressed in the malignant breast cell lines, and it was absent in normal breast epithelial cell lines. By western blotting, we were unable to distinguish between TPM1γ, λ, and ν protein expression, which were the only TPM1 gene protein isoforms potentially expressed. Some malignant cell lines demonstrated increased or decreased expression of these isoforms relative to the normal breast cell lines. Stress fiber formation did not correlate with TPM1γ RNA expression but significantly and inversely correlated with TPM1δ and TPM1λ expression, respectively. The exact differences in expression of these novel isoforms and their functional properties in breast epithelial cells will require further study.
对9种恶性乳腺上皮细胞系和3种正常乳腺细胞系进行了应激纤维形成以及TPM1亚型特异性RNA和蛋白质表达的检测。应激纤维形成在正常细胞系中较强(++++),在恶性细胞系中则有所不同(阴性至+++)。虽然TPM1γ和TPM1δ是TPM1的主要转录本,但没有明确证据表明存在TPM1δ蛋白表达。发现了4种新的人类TPM1基因RNA亚型(λ、μ、ν和ξ),在成人和胎儿的心脏组织中未鉴定到这些亚型。TPM1λ是在恶性乳腺细胞系中表达最频繁的亚型,在正常乳腺上皮细胞系中不存在。通过蛋白质免疫印迹法,我们无法区分TPM1γ、λ和ν的蛋白表达,这是仅有的可能表达的TPM1基因蛋白亚型。相对于正常乳腺细胞系,一些恶性细胞系中这些亚型的表达有所增加或减少。应激纤维形成与TPM1γ RNA表达无关,但分别与TPM1δ和TPM1λ表达显著负相关。这些新亚型在乳腺上皮细胞中的表达确切差异及其功能特性有待进一步研究。