Bcl-XL反义寡核苷酸增强食管癌细胞对5-氟尿嘧啶的敏感性

Bcl-XL antisense oligodeoxynucleotide sensitizes human esophageal cancer cell line to 5-fluorouracil

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 effects of the Bcl-XL antisense oligodeoxynucleotides （ASODN） in suppressing the Bel-XL expression and increasing the sensitivity of esophageal cancer cell line EC9706 to 5-fluorouracil （ 5-Fu ）, Methods The proliferation inhibitory rate of EC9706 was assessed by MTT, the expression of Bel-XL was detected by RT-PCR and Western blot, and the apoptotic changes were examined by acridine orange （AO） fluorescent staining and flow eytometry, respectively, Results In the group of ASODN combined with 5-Fu, the proliferation inhibitory rate of esophageal cancer cells was 71.58%, the expression inhibitory rate of Bel-XL mRNA was 81.25%, the expression of Bcl-XL protein was decreased significantly. The apoptosis rates detected by AO fluorescent staining and flow eytometry were 69.5% and （63.32±9.23 ）%, respectively. There were significant differences as compared with the cell control group, the vacuity control group, the N-ODN group, the ASODN group and the 5-Fu group, respectively （P 〈 0. 05 ）. Conclusion Bel-XL ASODN combined with 5-Fu can effectively inhibit the proliferation of esophageal cancer cells in vitro. Bel-XL ASODN can significantly increase the sensitivity of esophageal cancer cells to 5-Fu through suppressing the expression of Bel-XL.