本文選題：重組人精氨酸酶　切入點：人腦膠質瘤　出處：《中國臨床藥理學雜志》2017年02期

【摘要】：目的研究重組人精氨酸酶對人腦膠質瘤細胞U87的殺傷作用。方法培養(yǎng)人腦膠質瘤細胞U87,將對數(shù)生長期人腦膠質瘤細胞U87隨機分為12組:對照組僅加等量的培養(yǎng)液,其余11組實驗組以倍比稀釋的方式分別給予質量濃度為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,用甲基噻唑藍四氮唑鹽(MTT)法檢測各組人腦膠質瘤細胞U87細胞活力的變化;倒置顯微鏡觀察U87細胞經(jīng)重組人精氨酸酶作用后的形態(tài)學變化;Western blotting法檢測精氨酸琥珀酸合成酶(ASS)的表達。重組人精氨酸酶作用于人腦膠質瘤細胞U87時,分成補充L-精氨酸組和不補充L-精氨酸組,用MTT法檢測人腦膠質瘤細胞U87細胞活力的變化。結果不同質量濃度的重組人精氨酸酶作用U87細胞72 h后,U87細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)實驗組的細胞活力分別為75.12%,52.28%,44.09%,38.14%,差異有統(tǒng)計學意義(P0.05)。通過倒置顯微鏡觀察,重組人精氨酸酶作用U87細胞72 h后,0.2×10~(-2),1.6×10~(-2),1.25×10-1U·mL~(-1)實驗組的細胞密度隨著藥物濃度升高而顯著降低,鏡下可見U87細胞形態(tài)發(fā)生明顯的變形,皺縮,壞死。Western blotting實驗結果表明,U87細胞的ASS蛋白表達缺陷。當重組人精氨酸酶作用于U87細胞同時,補充部分L-精氨酸時的細胞活力為60.53%,未補充L-精氨酸組的細胞活力為47.48%,差異有統(tǒng)計學意義(P0.01)。結論由于人腦膠質瘤細胞U87細胞合成精氨酸的關鍵酶ASS蛋白表達缺陷,重組人精氨酸酶通過耗竭外源性精氨酸,對人腦膠質瘤細胞U87產(chǎn)生顯著的殺傷效應。
[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)科大學南方醫(yī)院藥物臨床試驗中心;
【基金】：國家自然科學基金資助項目(U1401226)
【分類號】：R739.41

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