Stat3反义寡脱氧核苷酸在体外诱导喉癌细胞凋亡

目的:探讨Stat3反义寡核苷酸对人喉癌Hep-2细胞株细胞凋亡的影响。方法:设计Stat3反义寡核苷酸序列,应用脂质体瞬时转染法转染人喉癌Hep-2细胞,应用Western blot和RT-PCR检测Stat3及p-Stat3的表达;MTT实验检测Stat3反义寡核苷酸对转染细胞的抑制情况,应用DNA ladder、吖啶橙/溴化乙啶(AO/EB)及流式细胞术检测细胞凋亡水平和形态变化。结果:Western blot和RT-PCR结果显示Stat3反义寡核苷酸分别在蛋白及mRNA水平显著抑制Stat3基因表达,并在蛋白水平抑制p-Stat3表达;转染Stat3反义寡核苷酸的喉癌细胞增殖明显受到抑制,与对照组比较差异有统计学意义(P<0.05);DNA ladder、AO/EB及流式细胞术证实Stat3反义寡核苷酸在体外可抑制Stat3基因表达并诱导人喉癌Hep-2细胞凋亡,且呈现浓度依赖性。结论:Stat3对喉癌细胞的过度生长、增殖起一定作用,Stat3反义寡核苷酸能够诱导喉癌细胞凋亡,抑制其增殖。

Apoptosis of laryngeal cacinoma cells induced by antisense oligodeoxynucleotides complementary Stat3 in vitro

Objective:To study the effects of oligodeoxynucleotides complementary Stat3 on apoptosis in laryngeal carcinoma Hep-2 cell. Method:Oligodeoxynucleotides complementary Stat3 was designed, which was transferred into laryngeal carcinoma Hep-2 cell by lipofection. Expression of Stat3 and p-Stat3 were detected by Western blot and PCR. MTT was used to observe the growth-inhibiting ratio. DNA ladder, AO/EB and FCM were used to observe the apoptosis of laryngeal carcinoma cell Hep-2 in vitro. Result:Western blot and PCR results demonstrated that oligodeoxynucleotides complementary Stat3 could significantly inhibit the expression of Stat3 and p-Stat3 in Hep-2 cell. MTT results showed that it could significantly suppress the growth of Hep-2 cell. The DNA ladder, AO/EB and FCM results showed it could inhibit the expression of Stat3 and induce the apoptosis of Hep-2 cell in a concentration-dependment manner. Conclusion:Oligodeoxynucleotides complementary Stat3 could induce the apoptosis and suppress cell proliferation in laryngeal carcinoma Hep-2 cell.