氯通道在5-Fu诱导的鼻咽癌细胞凋亡性细胞容积减小和凋亡中的作用

发布时间：2018-03-17 16:17

本文选题：氯通道　切入点：C1C-3　出处：《暨南大学》2011年硕士论文　论文类型：学位论文

【摘要】：目的：1)研究5-氟尿嘧啶(5-Fluorouacil,5-Fu)对低分化鼻咽癌细胞(CNE-2Z)凋亡的影响；2)5-Fu对CNE-2Z细胞氯通道的激活作用；3)氯通道在5-Fu诱导的CNE-2Z细胞凋亡性细胞容积减小和凋亡中的作用。方法：1)Hoechst染色、流式细胞术分析细胞凋亡；2)Scion Image图像分析软件测量细胞直径和面积,计算细胞标准容积；3)膜片钳全细胞记录技术记录5-Fu激活的CNE-2Z细胞全细胞电流,计算电流密度。结果： 1.5-Fu诱导CNE-2Z细胞产生明显的细胞凋亡,100μmol/L 5-Fu作用48 h的凋亡率为(49.2±3.2)%,氯通道阻断剂NPPB可使凋亡率下降至(12.5±2.6)%。 2.5-Fu诱导CNE-2Z细胞产生凋亡性细胞容积减小(Apoptotic volume decrease, AVD),胞外灌流100 umol/L 5-Fu 50 min细胞容积减小约9%,NPPB与5-Fu联合作用相同时间细胞仅皱缩1.5%；氯通道阻断剂明显抑制5-Fu诱导的AVD。 3.5-Fu可以激活CNE-2Z细胞产生一个电流特征与容积激活性氯通道相似的氯电流；氯通道阻断剂(NPPB, Tamoxifen)明显抑制5-Fu激活的电流。 4. ClC-3SiRNA阻断ClC-3氯通道表达,抑制5-Fu激活CNE-2Z细胞氯电流。结论： 1.阻断氯通道抑制5-Fu激活的CNE-2Z细胞氯电流,拮抗5-Fu诱发的凋亡性细胞容积减小和细胞凋亡等作用,提示氯通道在5-Fu诱导细胞凋亡中起重要作用。 2.ClC-3氯通道参与介导5-Fu激活的CNE-2Z细胞氯电流。
[Abstract]:Aim: to study the effect of 5-Fluorouaciline 5-Fu on apoptosis of poorly differentiated nasopharyngeal carcinoma (NPC) cell line CNE-2Z. The effect of 5-Fu on the activation of chloride channel in CNE-2Z cells and the role of chloride channel in 5-Fu induced apoptotic cell volume reduction and apoptosis of CNE-2Z cells were studied. Methods the cell diameters and area were measured by flow cytometry (FCM) and the cell diameter and area were measured by flow cytometry (FCM). The whole cell currents of 5-Fu activated CNE-2Z cells were recorded by patch clamp whole cell recording technique and the current density was calculated. Results:. 1. The apoptotic rate of CNE-2Z cells induced by 5-Fu was 49.2 卤3.2 for 48 h, and the apoptosis rate decreased to 12.5 卤2.6% by chloride channel blocker NPPB. 2.The volume of apoptotic cells in CNE-2Z cells induced by 5-Fu was reduced by Apoptotic volume decrease, AVDX, and extracellular perfusion of 100 umol/L 5-Fu 50 min cells. The cells only shrank at the same time after the combination of 5-Fu and 5-Fu, and the chloride channel blockers significantly inhibited 5-Fu induced AVDs. 3.5-Fu activated CNE-2Z cells to produce a chloride current similar to that of volume-activated chloride channel, and the chloride channel blocker NPPB( Tamoxifen) significantly inhibited the 5-Fu activated current. 4. ClC-3SiRNA blocked the expression of ClC-3 chloride channel and inhibited 5-Fu activation of chloride current in CNE-2Z cells. Conclusion:. 1. The blocking of chloride channel inhibits the chloride current of 5-Fu activated CNE-2Z cells, and antagonizes the effect of 5-Fu induced apoptosis on apoptotic cells, suggesting that chloride channel plays an important role in 5-Fu induced apoptosis. 2. ClC-3 chloride channel is involved in the regulation of chloride currents in 5-Fu activated CNE-2Z cells.
【学位授予单位】：暨南大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R739.63

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