IGF expression in HPV-related and HPV-unrelated human cancer cells.
IGF expression in HPV-related and HPV-unrelated human cancer cells.
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DOI：
10.3892/or.2014.3329
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发表时间：
2014-09
期刊：
影响因子：
4.2
通讯作者：
Barton E
中科院分区：
文献类型：
--
作者：
Durzyńska J;Barton E
人类IGF-1基因不仅产生胰岛素样生长因子-I(IGF-I)，还通过选择性剪接产生不同的羧基末端延伸，即E肽。我们和其他人已经证明了来源于IGF-1的人EB多肽(Heb)具有内在的生物学活性，并定位于转基因细胞的细胞核。由于Heb的作用可以补充IGF-I本身的活性，本研究的目的是在内源性蛋白和转录水平比较IGF-I在癌细胞系中的异构体，包括HeLa，U2OS，HepG2和K562细胞。以β-肌动蛋白为参照基因，用胰岛素样生长因子-1亚型特异性引物进行实时定量聚合酶链式反应。不同细胞系的IGF1转录水平不同，K562细胞表达水平(130.2±31.2)是U2OS细胞(1.7±1.1)的80倍左右。IGF-1b的相对拷贝数在HepG2(69.9±28.6)和K562细胞(28.3±6.7)中最高，而IGF-1a和IGF-1c的相对拷贝数在K562细胞中显著高于其他所有细胞系。用细胞总裂解物、胞浆和胞核部分进行免疫印迹，以确定IGF-I蛋白的水平和分布。K562细胞的总裂解物和核组分中的HEb含量最高，而细胞质组分中未见HEB。相反，IGF-IA在HeLa细胞中含量最高，且仅在胞浆部分富含。由于相对较低的IGF-1A转录水平和相对较高的原-IGF-1A蛋白水平是可能的，我们假设这些转录本可以更高的效率处理和/或蛋白产物可能是由病毒HPV癌基因在HeLa细胞中稳定的。我们认为，虽然分析IGF-1转录水平很重要，但在蛋白质水平上确定IGF亚型可能更有意义。
The human Igf-1 gene not only produces insulin-like growth factor-I (IGF-I), but also different carboxy-terminal extensions, known as E peptides, through alternative splicing. We and others have shown that human Eb peptide (hEb) derived from Igf-1 has intrinsic biological activity and is localized to nuclei of transfected cells. Since hEb actions can complement the activity of IGF-I itself, the aim of the present study was to compare IGF-I isoforms at the endogenous protein and transcript level in cancer cell lines, including HeLa, U2OS, HepG2 and K562 cells. Quantitative real-time PCR (qRT-PCR) using Igf-1 isoform specific primers was performed to determine expression patterns, using β-actin as a reference gene. The overall relative Igf-1 transcript level was different across the cell lines, with ~80-fold higher expression in K562 (130.2±31.2) than in U2OS cells (1.7±1.1). The relative copy number of Igf-1b was the highest in HepG2 (69.9±28.6) and K562 cells (28.3±6.7), whereas the relative copy numbers of Igf-1a and Igf-1c were significantly higher in K562 cells compared to all other cell lines. Immunoblotting using total cell lysates, cytoplasmic and nuclear fractions were carried out to determine the level and distribution of IGF-I proteins. K562 cells exhibited the highest level of hEb in total cell lysates and nuclear fractions and no cell lines displayed hEb in the cytoplasmic fractions. In contrast, IGF-IA was the highest in HeLa cells and was enriched only in the cytoplasmic fraction. Since relatively low IGF-1A transcript level but relatively high pro-IGF-1A protein level is plausible, we hypothesized that these transcripts could be processed with higher efficiency and/or the protein product may be stabilized by viral HPV oncogenes in HeLa cells. We assert that while it is important to analyze Igf-1 transcript level, it may be more relevant to determine the IGF isoforms at the protein level.
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影响因子：
11.2
作者：
Wahner Hendrickson AE;Haluska P;Schneider PA;Loegering DA;Peterson KL;Attar R;Smith BD;Erlichman C;Gottardis M;Karp JE;Carboni JM;Kaufmann SH
通讯作者：
Kaufmann SH
DOI：
10.1002/path.2608
发表时间：
2009-11
期刊：
The Journal of pathology
影响因子：
--
作者：
Mole S;McFarlane M;Chuen-Im T;Milligan SG;Millan D;Graham SV
通讯作者：
Graham SV
DOI：
10.1055/s-0032-1331699
发表时间：
2013-06
期刊：
Hormone and metabolic research = Hormon- und Stoffwechselforschung = Hormones et metabolisme
影响因子：
--
作者：
Durzyńska J;Wardziński A;Koczorowska M;Goździcka-Józefiak A;Barton ER
通讯作者：
Barton ER
DOI：
10.5114/wo.2012.28794
发表时间：
2012
期刊：
Contemporary oncology (Poznan, Poland)
影响因子：
--
作者：
Kasprzak A;Szaflarski W;Szmeja J;Andrzejewska M;Przybyszewska W;Koczorowska M;Drews M;Kaczmarek E
通讯作者：
Kaczmarek E
影响因子：
4.4
作者：
Klevebring, Daniel;Fagerberg, Linn;Lundeberg, Joakim
通讯作者：
Lundeberg, Joakim