阿苯达唑对耐顺铂肺癌细胞糖酵解、细胞周期及细胞凋亡的影响

Effects of albendazole on glycolysis,cell cycle and cell apoptosis of cisplatin-resistant lung cancer cells

目的研究阿苯达唑(ABZ)对人耐顺铂肺癌细胞(A549/DDP细胞)糖酵解、细胞周期及细胞凋亡的影响。方法用MTT比色法检测ABZ对A549/DDP细胞增殖的影响,求得抑制率为0%、25%、50%、75%所对应的ABZ浓度,并按抑制率将实验分为对照组,25%抑制浓度(IC25)组、半数抑制浓度(IC50)组和75%抑制浓度(IC75)组,每组给予相应浓度的ABZ处理,并按作用时间再将每组分为12、24、36h亚组。按照分组,在设定时间点,用比色法测定己糖激酶(HK)和丙酮酸激酶(PK)活性,酶标仪法测定乳酸脱氢酶(LDH)活性,RT-PCR法测定Akt和Myc mRNA表达,流式细胞仪检测细胞周期和凋亡。结果 ABZ抑制了A549/DDP细胞增殖,呈剂量依赖性,抑制率为0%、25%、50%、75%所对应的ABZ浓度分别为(0.00±0.00)μmol/L、(0.99±0.11)μmol/L、(5.73±0.65)μmol/L、(33.15±3.94)μmol/L。ABZ明显降低了A549/DDP细胞HK、PK、LDH的活性,且下调了Akt和Myc mRNA表达,细胞周期阻滞,凋亡明显。结论 ABZ能抑制A549/DDP细胞糖酵解酶活性,下调糖酵解相关基因表达,阻滞细胞周期,诱导细胞凋亡。

Objective To investigate effect of albendazole（ABZ）on glycolysis,cell cycle and cell apoptosis of human eisplatin-re- sistant lung cancer cells（A549/DDP cells）. Methods Effect of ABZ on proliferation of A549/DDP cells was detected by methyl thi- azolyl tetrazolium（MTT） colorimetry, then according to inhibition concentrations（IC）of 0%, 25%, 50 %, 75 % to devide cells into 4 groups-control group, IC25 group, IC50 group, IC75 group, moreover, based on ABZ action time to invide every group into 12,24 and 36 h sub-groups. At the setting points of action time, activitie of hexokinase （HK）, pyruvate kinase （PK）, dehydrogenase （LDH） were detected by colorimetry and ELIASA, expression of Akt and Myc mRNA were detected by RT-PCR, cell cycle and cell apopto- sis were detected by flow cytometry（FCM）. Results ABZ inhibited proliferation of A549/DDP cells in a dose-dependent manner, IC0 ,IC2s ,IC50 and IC75 respectively were（0. 00 ±0. 00）μmol/L, （0. 99 ±0. 11 ） tlmol/L, （5. 73 ±0. 65）/μmol/L, （33. 15 ± 3. 94） μmol/L. The activity of LDH, HK and PK of test groups decreased, expression of Akt mRNA and Myc mRNA was downregulated. Cell cycle was blocked,apoptosis was obvious. Conclusion ABZ can inhibit activity of glycolytic enzymes and expression of glycoly- sis-related genes,block cell cycle and induce apoptosis in vitro.