Repression of Transforming Growth Factor 1 Protein by Antisense Oligonucleotide-induced Increase of Adrenal Cell Differentiated Functions (*)
Repression of Transforming Growth Factor 1 Protein by Antisense Oligonucleotide-induced Increase of Adrenal Cell Differentiated Functions (*)
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通过反义寡核苷酸抑制转化生长因子 1 蛋白诱导的肾上腺细胞分化功能增加 (*)
DOI：
10.1074/jbc.271.18.11027
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发表时间：
1996
期刊：
影响因子：
--
通讯作者：
D. Langlois
中科院分区：
文献类型：
--
作者：
C. Le Roy;P. Leduque;P. Dubois;J. Saez;D. Langlois
转化生长因子β1 （tgf - β1）是牛肾上腺束状细胞（BAC）几种分化功能的有效抑制剂。此外，这些细胞表达和分泌这种因子。为了确定该肽是否在BAC中起自分泌作用，我们在细胞中转染了tgf - β1 mRNA翻译起始区互补的10 μM的未修饰义（SON）或反义（AON）寡核苷酸，试图抑制tgf - β1蛋白的合成。我们首先研究了p标记的tgf - β1 AON和SON的细胞摄取、稳定性和细胞内分布；第二，两种寡核苷酸对BAC特异性功能的影响。我们已经证明，在BAC中，tgf - β1 AON的摄取在转染8小时后达到平台（添加16%的放射性），并保持相当稳定至少24小时。相反，tgf - β1 SON的摄取在转染2小时后达到平台（添加8%的放射性），仅保持稳定3小时，然后下降。转染8 h后，再无寡核苷酸培养44 h，细胞内tgf - β1 AON水平仍然很高，约占放射性的8%，而tgf - β1 SON仅占1.2%。此外，AON存在于细胞质和细胞核部分，并在两个室中杂交。然而，tgf - β1 SON主要存在于未杂交的细胞质部分。tgf - β1 AON和SON均未改变tgf - β1 mRNA水平；然而，tgf - β1 AON引起细胞内tgf - β1免疫反应性消失，而SON不引起。最后，转染tgf - β1 AON的BAC的类固醇生成反应性增加了约2倍，这与细胞色素P450 17α-羟化酶和3β-羟基类固醇脱氢酶的mRNA水平增加了2倍有关。tgf - β1 SON和含有与tgf - β1 AON相同数量G核苷酸的打乱寡核苷酸对这些参数都没有任何影响。因此，这些研究表明tgf - β1对BAC分化功能具有自分泌抑制作用，而tgf - β1 AON可以克服这一作用。
Transforming growth factor β1 (TGFβ1) is a potent inhibitor of several differentiated functions in bovine adrenal fasciculata cells (BAC). In addition, these cells express and secrete this factor. To determine whether this peptide plays an autocrine role in BAC, cells were transfected with 10 μM unmodified sense (SON) or antisense (AON) oligonucleotide complementary to the translation initiation region of the TGFβ1 mRNA in an attempt to inhibit TGFβ1 protein synthesis. We investigated first, the cellular uptake, the stability, and the intracellular distribution of P-labeled TGFβ1 AON and SON; and second, the effects of both oligonucleotides on BAC specific functions. We have demonstrated that in BAC, the TGFβ1 AON uptake reached a plateau after 8 h of transfection (16% of the radioactivity added) and remained fairly constant for at least 24 h. In contrast, the uptake of TGFβ1 SON reached a plateau after 2 h of transfection (8% of the radioactivity added), remained stable for only 3 h, and then declined. After 8 h of transfection, followed by 44 h of culture without oligonucleotides, the intracellular level of TGFβ1 AON was still high with about 8% of the radioactivity added, whereas that of TGFβ1 SON represented only 1.2%. Moreover, AON was present in the cytoplasmic and nuclear fractions, and it was hybridized in both compartments. However, TGFβ1 SON was present mainly in the cytoplasmic fraction where it was not hybridized. Neither TGFβ1 AON nor SON modified TGFβ1 mRNA levels; however, TGFβ1 AON, but not SON, caused the disappearance of TGFβ1 immunoreactivity inside the cells. Finally, the steroidogenic responsiveness of BAC transfected with TGFβ1 AON increased about 2-fold, and this was associated with a 2-fold increase of the mRNA levels of both cytochrome P450 17α-hydroxylase and 3β-hydroxysteroid dehydrogenase. Neither TGFβ1 SON nor a scrambled oligonucleotide containing the same number of G nucleotides as TGFβ1 AON had any effect on these parameters. Thus, these studies demonstrate that TGFβ1 has an autocrine inhibitory effect on BAC differentiated functions, an effect that can be overcome by TGFβ1 AON.
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DOI：
10.1016/s0021-9258(18)98496-6
发表时间：
1991-08
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
J. Florini;K. Magri;D. Ewton;P. James;K. Grindstaff;P. Rotwein
通讯作者：
J. Florini;K. Magri;D. Ewton;P. James;K. Grindstaff;P. Rotwein
人体细胞中硫代磷酸同寡脱氧核苷酸的细胞药理学。
影响因子：
3.6
作者：
Gao,WY;Storm,C;Egan,W;Cheng,YC
通讯作者：
Cheng,YC
牛肾上腺皮质细胞色素 P-450(17 α)。
DOI：
--
发表时间：
1986
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Zuber,MX;John,ME;Okamura,T;Simpson,ER;Waterman,MR
通讯作者：
Waterman,MR
肾上腺皮质细胞中 3 β-羟基类固醇脱氢酶的调节：血管紧张素-II 和转化生长因子 β 的作用。
影响因子：
2.1
作者：
Rainey,WE;Naville,D;Mason,JI
通讯作者：
Mason,JI
转化生长因子β对绵羊肾上腺皮质细胞的影响。
影响因子：
4.1
作者：
Rainey,WE;Viard,I;Mason,JI;Cochet,C;Chambaz,EM;Saez,JM
通讯作者：
Saez,JM