STAT3反义寡核苷酸联合放疗对喉癌Hep-2细胞增殖、凋亡的影响

Effect of Hep-2 Cell Proliferation and Apoptosis by STAT3 Antisense Oligonucleotide and Radiotherapy in Laryngeal Cancer Cells

目的:观察STAT3反义寡核苷酸(STAT3 AS-ON)联合放疗对喉癌Hep-2细胞增殖、凋亡的影响。方法:体外培养Hep-2细胞,应用STAT3 AS-ON转染Hep-2细胞后,将培养细胞按是否进行放疗分为未放疗组与放疗组,未放疗组根据转染复合物的不同分为反义组、正义组及空白对照组,而放疗组分为反义+放疗组、正义+放疗组及放疗对照组。根据转染复合物的浓度(100、200、400nmol/L)又将对应各组分为反义100、反义200、反义400、正义100、正义200、正义400组。其中放疗组Hep-2细胞给予γ射线(5Gy)照射。MTT法检测细胞增殖情况,流式细胞术检测细胞凋亡率。结果:反义+放疗组的细胞存活率明显低于正义+放疗组和放疗对照组(P<0.01),且随着STAT3 AS-ON转染浓度的增加而降低,反义+放疗组的细胞凋亡率明显高于正义+放疗组和放疗对照组(P<0.01),而正义+放疗组和放疗对照组相比差异无显著性意义(P>0.05)。结论:STAT3 AS-ON联合γ射线作用于Hep-2细胞后,细胞增殖抑制和诱导凋亡作用明显增强,表明选择性阻断细胞内STAT3信号转导通路联合放疗可能为喉癌的综合治疗提供新的途径。

Objective：To observe the effect of STAT3 antisense oligonueleotide in combination with irradiation of γ-rays on proliferation and apoptosis of Hep-2 cells in human laryngeal cancer. Methods ： Hep-2 cells were cultured in vitro after Hep-2 cells was transfected with STAT3 AS-ON and all the cultured cells were divided into two-groups about the radiotherapy group and the non-radiotherapy group according to whether they would be irradiated byγ-rays or not. According to the difference of the transfection complex,the cells of the non-radiotherapy group were divided into antisense group,sense group and blank control group,while the cells of the radiotherapy group were divided into antisense ＋ radiotherapy group,sense ＋ radiotherapy group and radiotherapy control group. According to the difference of the concentration of the transfection complex 100,200,400 nmol/L, the groups were divided into antisense 100, antisense 200, antisense 400, sense 100, sense 200and sense 400 group. The radiotherapy group was irradiated by γ-rays （5 Gy）, The cell proliferation was assayed by MTT. Flow cytometry was performed to analyze apoptosis. Results：The survival rate of the Hep-2 cells in the antisense ＋ radiotherapy group was significantly lower than that of the sense ＋ radiotherapy and radiotherapy control group（ P 〈 0, 01 ） ,and was in invere proportion with the concentration of STAT3 AS- ON ,and the rate of apoptosis was significantly higher than that of the sense ＋ radiotherapy and radiotherapy control group（ P 〈 0.01 ）, while there was no significant difference between the sense ＋ radiotherapy and radiotherapy control group （P 〉 0.05 ）. Conclusion-STAT3 AS-ON and γ-rays can suppress the growth and induce the apoptosis rate if they act on Hep-2 cells at one time. Selective inhibition of STAT3 signaling pathway may provide a new therapeutic approach to combined treatment of laryngeal cancers.