本文關鍵詞：肝素對KGF-2介導的NIH3T3增殖及信號轉導機制研究　出處：《吉林農(nóng)業(yè)大學》2011年碩士論文　論文類型：學位論文

  更多相關文章： KGF-2 肝素 NIH3T3細胞 信號通路 ERK

【摘要】：KGF-2是一種非常具有潛力的治療分子,能夠特異性促進細胞遷移、細胞增殖、細胞分化；在脊椎動物的多種組織和器官發(fā)育中起重要調(diào)節(jié)作用。KGF-2具有兩種酪氨酸激酶受體FGFR1和FGFR2, FGFR1是其低親和力受體,FGFR2是高親和力受體。NIH3T3細胞是一種上皮細胞,屬成纖維細胞一種,它的表面含有FGFR1,因此,KGF-2能夠與其表面受體結合,使受體發(fā)生自身磷酸化,將信號傳遞到細胞內(nèi),實現(xiàn)信號傳遞。 肝素/硫酸肝素類自身具有多種生物學功效,如抗凝、抗炎等,這些生物學功效需要與多種蛋白質(zhì)相互作用來發(fā)揮,以往實驗表明多糖是生長因子信號傳遞的必需分子。 本實驗中首先應用生物學活性檢測的常規(guī)方法MTT法,研究KGF-2對NIH3T3細胞的促增殖作用,其次,探索了肝素對KGF-2發(fā)揮生物學活性起到的調(diào)控作用,證明肝素能夠促進KGF-2與FGFR1的結合,促進NIH3T3細胞的增殖,最佳肝素濃度為200μg/ml,能夠顯著增強KGF-2對NIH3T3細胞增殖的促進作用。然后,建立應用NIH3T3測定KGF-2生物學活性的新方法,并優(yōu)化了活性測定的條件,最優(yōu)條件為：細胞接種密度為8000,基礎培養(yǎng)基中血清濃度為0.5%,藥物作用時間為48h,最佳藥物濃度為100μg/m1,藥物儲存時盡量避免反復凍融,藥物緩沖體系選擇磷酸鹽緩沖體系。最后,對ERK在KGF-2促進NIH3T3增殖時的作用進行研究,胞外信號調(diào)節(jié)激酶途徑是涉及調(diào)節(jié)細胞生長信號網(wǎng)絡的核心,對ERK磷酸化水平進行研究,結果表明KGF-2在促進NIH3T3增殖時p-ERK有表達,啟動ERK途徑,最佳磷酸化時間是KGF-2作用15min；然后加入ERK信號通路抑制劑PD98059,再檢測p-ERK水平,結果顯示加入抑制劑后ERK磷酸化水平被抑制,證明此增殖實驗中所啟動的信號通路為ERK途徑。 總之,本實驗證明肝素能夠調(diào)控KGF-2對NIH3T3細胞的增殖作用,建立了應用NIH3T3細胞測定KGF-2生物學活性的新方法,并優(yōu)化了活性測定條件,對KGF-2發(fā)揮作用時的ERK信號通路進行研究,為KGF-2在臨床治療研究奠定了基礎。
[Abstract]:KGF - 2 is a very potential therapeutic molecule capable of specifically promoting cell migration , cell proliferation , cell differentiation , and plays an important role in the development of various tissues and organs of vertebrates . KGF - 2 has two tyrosine kinase receptors FGFR1 and FGFR2 , FGFR1 is a low - affinity receptor , FGFR2 is a high - affinity receptor . The heparin / heparan sulfate class itself has a variety of biological effects , such as anticoagulation , anti - inflammation , etc . These biological effects need to interact with a variety of proteins , and previous experiments have shown that polysaccharides are essential molecules for signal transduction of growth factors . The effects of KGF - 2 on the proliferation of NIH 3T3 cells were studied by MTT assay . The effect of KGF - 2 on the proliferation of NIH 3T3 cells was investigated . The optimal conditions were as follows : cell inoculation density was 8000 , the optimal concentration of heparin was 200 渭g / ml , the optimal concentration of heparin was 100 渭g / ml . In conclusion , this experiment proves that heparin can regulate the proliferation of KGF - 2 to NIH 3T3 cells , establish a new method to measure the biological activity of KGF - 2 by using NIH 3T3 cells , optimize the activity assay condition , and study the ERK signal pathway when KGF - 2 plays a role , which lays a foundation for the clinical treatment of KGF - 2 .

【學位授予單位】：吉林農(nóng)業(yè)大學
【學位級別】：碩士
【學位授予年份】：2011
【分類號】：R363

【引證文獻】

相關碩士學位論文 前1條

1 高麗昌;KGF-2對角膜基質(zhì)細胞的修復作用及其相關機制研究[D];吉林農(nóng)業(yè)大學;2012年

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本文編號：1395510