沅錞 asked in 健康其他:保健 · 1 decade ago

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Identification of signaling pathways of IGF-I mediated COX-2 expression in pancreatic cancer cells. (a) Western blot analysis for COX-2. Cells were pretreated with signaling inhibitors to MAPK/Erk-1/2 (UO126), or PI-3K/Akt (wortmannin, WT), as described in Section 2. Cells were subsequently stimulated for 4 h by adding rhIGF-I. IGF-I treatment resulted in a 5-fold increase in COX-2 protein expression, which was selectively mediated by the MAPK (Erk-1/2) signaling pathway. Inhibition of PI-3K/Akt signaling did not reduce COX-2 expression.

(b) COX-2 expression was investigated following treatment with inhibitors to PI-3K (LY), P38 (SB) and SAPK (SP). IGF-I mediated COX-2 induction was not blunted by pretreatment with these inhibitors prior to IGF-I exposure.

(c) Northern blot analysis for COX-2 mRNA expression. Cells were pretreated with inhibitors as described above, except that cells were treated for 1 h with rhIGF-I. Stimulation with rhIGF-I increased COX-2 mRNA in control cells, whereas this response was blunted after inhibition of MAPK/Erk-1/2 by UO126. In addition, constitutive COX-2 mRNA expression was lowered by 70% upon treatment with UO126. In contrast, inhibition of PI-3K/Akt by LY did not reduce constitutive or inducible COX-2 mRNA expression.

(d) Effect of HIF-1a inhibition on COX-2 expression. L3.6pl cells were transiently transfected with RNAi for HIF-1a. After 24 h, cells were exposed to rhIGFI. RNAi to HIF-1a markedly reduced constitutive COX-2 expression without altering the inducible response to IGF-I. b-Actin served as a loading control.

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  • 小君
    Lv 6
    1 decade ago
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    鑑定信號轉導途徑中IGF - I介導的COX - 2的表達在胰腺癌細胞。 (一)免疫印跡分析為環氧合酶-2 。細胞預處理信號抑製劑,以mapk/erk-1/2 ( uo126 ) ,或pi-3k/akt ( Wortmannin對,九倉) ,描述在第2條。細胞隨後刺激4 h的加入rhigf我。胰島素樣生長因子I治療,導致1 5倍增加,環氧合酶-2蛋白的表達,這是有選擇性地介導由蛋白激酶( ERK的- 1 / 2 )信號轉導通路。抑制pi-3k/akt信號並未減少COX - 2的表達。

    (二) COX - 2的表達,考察了以下治療與抑製劑,以丕的3 K ( ly ) , P38的(銻)和sapk ( SP )的。中IGF - I介導的環氧合酶-2的誘導是不會減弱預處理與這些抑製劑之前, IGF - I的接觸。

    (三) Northern blot分析為COX - 2 mRNA的表達。細胞預處理抑製劑如上文所述,除了細胞治療1 h後與rhigf我。刺激rhigf我增加了環氧合酶-2 mRNA在控制細胞,而這種反應是減弱後,抑制mapk/erk-1/2由uo126 。此外,本構環氧合酶-2 mRNA的表達,降低了70 %後的待遇與uo126 。對比之下,抑制pi-3k/akt由ly並未減少構或誘導COX - 2 mRNA的表達。

    (四)效果的HIF - 1的抑制COX - 2的表達。 l3.6pl細胞瞬時轉染的RNAi為中HIF - 1 。 24 h後,細胞暴露於rhigfi 。 RNAi技術,以中HIF - 1顯著減少構COX - 2的表達,而不改變誘導的反應中IGF - I 。的B -肌動充當加載控制。

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