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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。
Objective: To investigate the effect of Stat3 antisense oligonucleotide on apoptosis of human laryngeal carcinoma Hep-2 cell line. Methods: Stat3 antisense oligonucleotides were designed and transfected into human laryngeal carcinoma Hep-2 cells by lipofectamine. The expressions of Stat3 and p-Stat3 were detected by Western blot and RT-PCR. Stat3 Antisense oligonucleotides on the transfected cells, DNA ladder, acridine orange / ethidium bromide (AO / EB) and flow cytometry were used to detect apoptosis and morphological changes. Results: The results of Western blot and RT-PCR showed that Stat3 antisense oligonucleotide significantly inhibited Stat3 gene expression at protein and mRNA levels and inhibited p-Stat3 protein expression at the protein level. Stat3 antisense oligodeoxynucleotides transfection The proliferation of cancer cells was significantly inhibited (P <0.05). DNA ladder, AO / EB and flow cytometry confirmed that Stat3 antisense oligonucleotides inhibited the expression of Stat3 gene in vitro Human laryngeal carcinoma Hep-2 cells apoptosis in a concentration-dependent manner. Conclusion: Stat3 may play an important role in the overgrowth and proliferation of laryngeal carcinoma cells. Stat3 antisense oligonucleotide can induce the apoptosis of laryngeal carcinoma cells and inhibit its proliferation.