本文選題：重組人精氨酸酶　切入點(diǎn)：人腦膠質(zhì)瘤　出處：《中國(guó)臨床藥理學(xué)雜志》2017年02期

【摘要】：目的研究重組人精氨酸酶對(duì)人腦膠質(zhì)瘤細(xì)胞U87的殺傷作用。方法培養(yǎng)人腦膠質(zhì)瘤細(xì)胞U87,將對(duì)數(shù)生長(zhǎng)期人腦膠質(zhì)瘤細(xì)胞U87隨機(jī)分為12組:對(duì)照組僅加等量的培養(yǎng)液,其余11組實(shí)驗(yàn)組以倍比稀釋的方式分別給予質(zhì)量濃度為0.2×10~(-3),0.5×10~(-3),0.1×10~(-2),0.2×10~(-2),0.4×10~(-2),0.8×10~(-2),1.6×10~(-2),3.2×10~(-2),6.3×10~(-2),1.25×10-1,0.25 U·mL~(-1)的重組人精氨酸酶,用甲基噻唑藍(lán)四氮唑鹽(MTT)法檢測(cè)各組人腦膠質(zhì)瘤細(xì)胞U87細(xì)胞活力的變化;倒置顯微鏡觀察U87細(xì)胞經(jīng)重組人精氨酸酶作用后的形態(tài)學(xué)變化;Western blotting法檢測(cè)精氨酸琥珀酸合成酶(ASS)的表達(dá)。重組人精氨酸酶作用于人腦膠質(zhì)瘤細(xì)胞U87時(shí),分成補(bǔ)充L-精氨酸組和不補(bǔ)充L-精氨酸組,用MTT法檢測(cè)人腦膠質(zhì)瘤細(xì)胞U87細(xì)胞活力的變化。結(jié)果不同質(zhì)量濃度的重組人精氨酸酶作用U87細(xì)胞72 h后,U87細(xì)胞的活力均降低且呈明顯的劑量依賴性,0.2×10~(-2),0.8×10~(-2),1.6×10~(-2),0.25 U·mL~(-1)實(shí)驗(yàn)組的細(xì)胞活力分別為75.12%,52.28%,44.09%,38.14%,差異有統(tǒng)計(jì)學(xué)意義(P0.05)。通過倒置顯微鏡觀察,重組人精氨酸酶作用U87細(xì)胞72 h后,0.2×10~(-2),1.6×10~(-2),1.25×10-1U·mL~(-1)實(shí)驗(yàn)組的細(xì)胞密度隨著藥物濃度升高而顯著降低,鏡下可見U87細(xì)胞形態(tài)發(fā)生明顯的變形,皺縮,壞死。Western blotting實(shí)驗(yàn)結(jié)果表明,U87細(xì)胞的ASS蛋白表達(dá)缺陷。當(dāng)重組人精氨酸酶作用于U87細(xì)胞同時(shí),補(bǔ)充部分L-精氨酸時(shí)的細(xì)胞活力為60.53%,未補(bǔ)充L-精氨酸組的細(xì)胞活力為47.48%,差異有統(tǒng)計(jì)學(xué)意義(P0.01)。結(jié)論由于人腦膠質(zhì)瘤細(xì)胞U87細(xì)胞合成精氨酸的關(guān)鍵酶ASS蛋白表達(dá)缺陷,重組人精氨酸酶通過耗竭外源性精氨酸,對(duì)人腦膠質(zhì)瘤細(xì)胞U87產(chǎn)生顯著的殺傷效應(yīng)。
[Abstract]:Objective to study the killing effect of recombinant human argininase on human glioma cell U87. Methods Human glioma cell U87 in logarithmic phase was randomly divided into 12 groups: the control group was treated with the same amount of culture medium. The other 11 experimental groups were given the recombinant human argininase with a mass concentration of 0.2 脳 10 ~ (-10) ~ (-3) and 0.5 脳 10 ~ (-1) U ~ (-1). The activity of U87 cells in each group was detected by MTT method. The activity of U87 cells in each group was detected by MTT- (0. 8 脳 10 ~ (-2)) 脳 10 ~ (-10) ~ (2) ~ (3) 脳 10 ~ (-10) ~ (2) ~ (2). The morphological changes of U87 cells treated with recombinant human argininase were observed by inverted microscope. The expression of arginine succinate synthase (ASS) in U87 cells was detected by Western blotting. Divided into L-arginine supplementation group and non-L-arginine supplementation group, The activity of U87 cells was detected by MTT assay. Results the activity of U87 cells decreased in a dose-dependent manner after 72 h treatment with different concentrations of recombinant human argininase. The activity of U87 cells decreased in a dose-dependent manner. The cell viability was 75.12 and 52.28 and 44.09 and 38.14, respectively. The difference was statistically significant (P 0.05). After 72 hours of treatment with recombinant human argininase, the cell density of U87 cells decreased significantly with the increase of drug concentration, and U87 cells were morphologically deformed and shrinked under microscope. The results of Western blotting assay showed that the expression of ASS protein was deficient in U87 cells. The cell viability of L- arginine supplementation group was 60.53, and that of non-L-arginine group was 47.48. The difference was statistically significant (P 0.01). Conclusion the key enzyme ASS protein expression of arginine synthesis in human glioma cell U87 is defective. Recombinant human argininase exerts significant killing effect on human glioma cell line U87 by exhausting exogenous arginine.
【作者單位】： 南方醫(yī)科大學(xué)南方醫(yī)院藥物臨床試驗(yàn)中心;
【基金】：國(guó)家自然科學(xué)基金資助項(xiàng)目(U1401226)
【分類號(hào)】：R739.41

【相似文獻(xiàn)】

相關(guān)期刊論文 前1條

1 岳瑛;岳冀;鄧紅燕;付峰;漆家學(xué);張琪;;重組長(zhǎng)效人精氨酸酶抑制腫瘤細(xì)胞生長(zhǎng)的研究[J];武警醫(yī)學(xué);2010年03期

相關(guān)碩士學(xué)位論文 前1條

1 岳瑛;人精氨酸酶與人的血清蛋白融合蛋白在畢赤酵母中的表達(dá)及活性測(cè)定[D];第四軍醫(yī)大學(xué);2010年

，

本文編號(hào)：1662096