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目的探讨Bcl -XL反义寡核苷酸(ASODN)在下调Bcl- XL表达、增加食管癌细胞株EC9706对5氟尿嘧啶(5Fu)敏感性中的作用。方法设细胞对照组、空白对照组、无关序列寡核苷酸(N ODN)组、ASODN组、5Fu组及ASODN联合5Fu组,应用四甲基偶氮唑蓝(MTT)法检测EC9706细胞的增殖抑制率;RT PCR和Westernblot检测Bcl- XLmRNA和蛋白表达水平的改变;吖啶橙荧光染色和流式细胞技术定量检测EC9706细胞的凋亡率。结果Bcl -XLASODN联合5Fu治疗组对食管癌细胞增殖的抑制率为71.58%,对Bcl XLmRNA表达抑制率为81.25%,并可显著下调Bcl XL的蛋白表达;凋亡指数为69.5%,凋亡率为(63.32±9.23)%,与细胞对照组、空白对照组、N -ODN组、ASODN组及5Fu组比较,差异均有统计学意义(P<0.05)。结论ASODN联合5Fu可有效抑制食管癌细胞EC9706的增殖,通过ASODN下调Bcl XL表达,可显著增强食管癌细胞对5Fu的敏感性。
Objective To investigate the role of Bcl-XL antisense oligonucleotide (ASODN) in down-regulating Bcl-XL expression and increasing the sensitivity of EC9706 cells to 5-fluorouracil (5Fu). Methods The proliferation of EC9706 cells was detected by MTT assay in cell control group, blank control group, N ODN group, ASODN group, 5Fu group and ASODN combined with 5Fu group. The inhibitory rate of Bcl-XL mRNA and protein expression was detected by RT PCR and Western blot. Apoptosis rate of EC9706 cells was quantitatively detected by acridine orange fluorescence staining and flow cytometry. Results The inhibitory rate of Bcl-XLASODN combined with 5Fu treatment group was 71.58%, the inhibitory rate of Bcl XLmRNA expression was 81.25%, and the protein expression of Bcl XL was significantly decreased. The apoptotic index was 69.5% (63.32 ± 9.23)%, compared with the cell control group, blank control group, N-ODN group, ASODN group and 5Fu group, the difference was statistically significant (P <0.05). Conclusions ASODN combined with 5Fu can effectively inhibit the proliferation of esophageal cancer cells EC9706. The down-regulation of Bcl XL expression by ASODN can significantly enhance the sensitivity of esophageal cancer cells to 5Fu.