阿霉素联合二甲双胍应用对舌癌CAL27细胞增殖及凋亡的影响

发布时间：2018-11-16 10:47

【摘要】：目的:研究阿霉素(ADM)与二甲双胍(MET)单独应用及二者联合用药对体外培养的舌癌CAL27细胞增殖以及凋亡的影响。方法:1.依次采用MET药物浓度5mmol/L、10mmol/L、20mmol/L、40mmol/L、80mmol/L、160mmol/L。ADM药物浓度0.02mg/L、0.04mg/L、0.08mg/L、0.16mg/L、0.32mg/L。分别对体外培养的舌癌CAL27细胞作用24h、48h、72h后,用CCK-8法检测各用药组的吸光度值,算出细胞的抑制率,利用SPSS 17.0软件计算出两组药物单独使用时抑制50%细胞生长的浓度IC50;而联合用药组则采取单独用药时所得的ADM组IC50+MET组IC50处理CAL27细胞48h后上酶标仪检测吸光度值,计算出联合用药组的细胞抑制率;2.根据CCK-8实验组数据,选取48h MET半抑制浓度10mmol/L,ADM半抑制浓度0.05mg/L及二者联合作用于CAL27细胞后,用流式细胞术检测各实验组细胞凋亡的情况;3.采用q RT-PCR技术检测MET 10mmol/L,ADM 0.05mg/L及MET 10mmol/L+ADM 0.05mg/L联合应用48h后舌癌CAL27细胞的相关凋亡基因Bax及Bcl-2 m RNA相对表达的情况。结果:1.CCK-8数据显示MET与ADM单独作用24h、48h、72h后,均对舌癌CAL27细胞的增殖产生抑制作用,并且在一定浓度范围内呈剂量与时间的依赖性,二者联合应用时比单独用药对CAL27细胞的增殖抑制作用更加明显。2.流式细胞术结果显示经MET 10mmol/L+ADM 0.05mg/L联合用药后比单独药物组更能促进细胞的凋亡。3.q RT-PCR显示MET及ADM二者合用时舌癌CAL27细胞的抗细胞凋亡基因Bcl-2的相对表达水平与单独用药组相比下调趋势更明显,而促细胞凋亡基因Bax m RNA的相对表达与单独用药组相比上调趋势更明显。结论:二甲双胍作为降血糖药物与阿霉素作用相似,单独使用时具有对舌癌CAL27细胞体外增殖的抑制作用。当二者联合应用时,对舌癌CAL27细胞的增殖抑制及促进凋亡的效果比单独用药更为显著。
[Abstract]:Aim: to study the effects of adriamycin (ADM) and metformin (MET) on the proliferation and apoptosis of tongue cancer CAL27 cells cultured in vitro. Methods: 1. The drug concentration of MET was 5 mmol / L 10 mmol / L and 20 mmol / L = 40 mmol / L = 80 mmol / L = 160 mmol / L = 0.02 mg / L 0.04 mg / L 0.08 mg / L = 0.16 mg / L = 0.32 mg / L. After cultured tongue cancer CAL27 cells were treated for 24 h or 48 h for 72 h, the absorbance of each drug group was measured by CCK-8 method, and the inhibition rate of the cells was calculated. SPSS 17.0 software was used to calculate the concentration of IC50; that inhibited 50% cell growth in two groups of drugs alone. However, in the combined treatment group, the CAL27 cells in the IC50 MET group were treated with IC50 for 48 hours, and the absorbance value was measured by enzyme labeling instrument, and the cell inhibition rate of the combined treatment group was calculated. 2. According to the data of CCK-8 experimental group, 10 mmol / L MET semi-inhibitory concentration (0.05mg/L) of 48 h and their combination were used to detect the apoptosis of CAL27 cells by flow cytometry. 3. Q RT-PCR technique was used to detect the relative expression of Bax and Bcl-2 m RNA in CAL27 cells of tongue cancer treated with MET 10 mmol / L 0.05mg/L and MET 10mmol/L ADM 0.05mg/L for 48 h. Results: 1.CCK-8 data showed that the proliferation of tongue cancer CAL27 cells was inhibited by MET and ADM alone for 24 h or 48 h or 72 h, and in a dose-dependent and time-dependent manner. The inhibitory effect of the two drugs on the proliferation of CAL27 cells was more obvious than that of the drug alone. 2. 2. The results of flow cytometry showed that MET 10mmol/L ADM 0.05mg/L combined with drugs could promote cell apoptosis more than that of single drug group. 3. Q RT-PCR showed antiapoptotic genes in CAL27 cells of tongue cancer combined with MET and ADM. The relative expression level of Bcl-2 was more down-regulated than that of the control group. The relative expression of apoptosis-promoting gene Bax m RNA was significantly higher than that of the control group. Conclusion: metformin, as a hypoglycemic drug, can inhibit the proliferation of tongue cancer CAL27 cells in vitro. When combined, the effect of inhibiting proliferation and promoting apoptosis in CAL27 cells of tongue cancer was more significant than that of drug alone.
【学位授予单位】：广西医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R739.86

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