应用免疫细胞化学和原位杂交技术光镜下检测培养的肝癌Hep G2细胞中α -胰蛋白酶间链抑制剂及其不同mrna。

The Histochemical Journal Pub Date : 1994-03-01
H Borghi, A Callé, R Sesboüé, J Bourguignon, M Diarra-Mehrpour, J P Martin
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引用次数: 0

摘要

hepg2肝癌细胞系合成α -胰蛋白酶抑制剂(ITI)。该蛋白酶抑制剂和该家族的其他蛋白包括四条多肽链:三条重链(HC1, HC2, HC3)和一条轻链(bikunin)。在本研究中,我们已经通过免疫荧光证明ITI主要在核周细胞质区检测到,与白蛋白或α -1-抗胰蛋白酶类似。在非同步培养的所有Hep G2细胞中都存在这四个多肽链的mrna,已通过原位杂交证明。通过对标记的分析,对四个ITI基因的转录进行了评估,结果表明,来自轻链的mrna明显高于来自重链的mrna。HC2链对应的mrna比HC1和HC3链对应的mrna更有代表性。在培养的Hep G2细胞中,基于原位杂交技术的mrna定量显示,它们的相对数量从大到小依次为L、HC2、HC3和HC1。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Light microscopical detection of inter-alpha-trypsin inhibitor and its different mRNAs in cultured hepatoma Hep G2 cells using immunocytochemical and in situ hybridization techniques.

The Hep G2 hepatoma cell line synthesizes the inter-alpha-trypsin inhibitor (ITI). This protease inhibitor and the other proteins of this family include four polypeptides chains: three heavy chains (HC1, HC2, HC3) and one light chain (bikunin). In the present study, we have demonstrated by immunofluorescence that ITI is detected mainly in perinuclear cytoplasmic zones comparable to those of albumin or alpha-1-antitrypsin. The presence of the mRNAs of the four polypeptide chains in all Hep G2 cells of a non-synchronized culture have been demonstrated by in situ hybridization. An evaluation of the transcription of the four ITI genes through an analysis of markings brings to the fore a clearly much higher rate of mRNAs from the light chain than from the heavy chains. The mRNAs corresponding to the HC2 chains are more heavily represented than are those corresponding to the HC1 and HC3 chains. In Hep G2 cells in culture, a quantification of mRNAs based on the in situ hybridization technique shows that their relative quantities, in decreasing order, are those of L, HC2, HC3 and HC1.

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