Bcl-xL反义寡核苷酸与博莱霉素协同抑制鳞状细胞癌细胞增殖的研究

The Effect of Bcl-xL Antisense Oligonucleotides Combined with Bleomycin on Growth Inhibition in Squamous Cell Carcinoma Tca8113 Cells

目的:研究bcl-xl反义寡核苷酸(ASODN)与博莱霉素(BLM)联用对口腔鳞状细胞癌细胞(Tca8113)增殖的影响及其作用机理。方法:设计合成bcl-xl的ASODN在脂质体的介导下转染Tca8113细胞,设无义寡核苷酸组(NSODN)和空白对照组进行比较。48h后博莱霉素作用于转染后的细胞。以荧光原位末端缺口标记法检测bcl-xl反义寡核苷酸与博莱霉素诱导癌细胞凋亡的作用机理,流式细胞仪检测细胞凋亡率。结果:bcl-x1反义寡核苷酸与博莱霉素的作用机理为激活细胞的内源性核酸内切酶,使核小体内DNA断裂出现缺口,产生凋亡。ASODN组、BLM组和ASODN+BLM组的凋亡率明显高于NSODN组和空白对照组(P<0.05)。ASODN+BLM组的凋亡率明显高于ASODN组和BLM组(P<0.05)。结论:Bcl-xL反义寡核苷酸与博莱霉素能协同抑制口腔鳞状细胞癌细胞的增殖。

Objective： To study the mechanism and effect of bcl--xl antisense oligonucleotides combined with bleo- mycin on the growth of squamous cell carcinoma cell line Tca8113. Methods： ASODN of target bcl xl was synthe- sized and transfected to Tca8113 cells through lipofectin vectin. At the same time, positive control （nonsense oligonueleotides,NSOON） and nomal control were set for comparison. After 48h the cells were treated with bleomycin. TdT--mediated dUTP nick end labeling assay （TUNEL） was used to study the mechanism of bcl--xl ASODN combined with hleomycin in inducing apoptosis in Tca8113 cells. Flow cytometry （FCM） was used for the detection of apoptosis rate. Results： The mechanism was that bci--xl ASODN combined with bleomycin activated the endoge- nous endonuclease in the Tca8113 cells, which broke the DNA in the nuhody, inducing apoptosis. FCM analysis showed that apoptosis rates in ASODN group ,BLM group and ASODN＋BLM group were significantly higher than those in control groups （P〈0.05）. The apoptosis rate in ASODN＋BLM group was significantly higher than in ASODN group and BLM group （P〈0.05）. Conclusion： The bcl--xl ASODN combined with bleomycin can enhance the inhibitory effect on squamous cell carcinoma cell.