Bax、Bcl-2ASODN联合转染增强宫颈癌HeLa细胞放射敏感性的实验研究

Study on enhancing radiosensitivity to HeLa cells by combination genes transfection of Bax and Bcl-2 ASODN.

目的:研究凋亡抑制基因bcl-2反义寡核苷酸(ASODN)和促凋亡基因bax联合转染对宫颈癌HeLa细胞凋亡的影响,进而探求其放射增敏作用。方法:用逆转录病毒颗粒感染HeLa细胞,获得HeLa-bax细胞后,将bcl-2ASODN转染HeLa及HeLa-bax细胞,36h后放射治疗,通过MTT法、细胞形态学及流式细胞仪检测细胞增殖与凋亡,半定量RT-PCR测bcl-2及bax基因的表达。结果:(1)HeLa细胞经bcl-2ASODN及bax联合转染并放疗后,光镜观察有明显凋亡趋势;(2)MTT法及流式细胞仪检测其凋亡率明显高于单基因转染+放疗组(P<0.05);(3)RT-PCR结果显示,bax基因表达显著增强,而bcl-2基因表达较各对照组明显减弱。结论:凋亡抑制基因bcl-2ASODN和促凋亡基因bax联合转染诱导HeLa细胞凋亡效果明显优于单基因转染,可有效提高HeLa细胞的放射敏感性。

Objective：To investigate the apoptotic effect of human cervical cancer cell lines by transfecting antiapoptosis gene-bcl-2 ASODN and pro-apoptotic gene-bax and further explore the value of combined genes transfection on enhancing the radiosensitivity to HeLa cells. Methods： pLXSN-bax was transfected into HeLa cells using retroviral granules, and then bcl-2ASODN was transfected into normal cells and HeLa-bax cells. After 36h, cells were radiotherapied and continued to be cultured for 3 days. The MTT, morphology and flow cytometry were used to detect the proliferation and apoptosis. RT-PCR was used to identify the mRNA levels of bcl-2 and bax. Results： （ 1 ） After genes transfection and radiotherapy a significant apoptosis was observed by microscopy; （2）The inhibition ratio and apoptosis of cells treated with genes transfection and radiotherapy were significantly higher than those of other groups （P 〈 0. 05 ）;（3 ）Compared with other groups,RT-PCR showed the bax gene＇s expression was significantly enhanced while the expression of bcl-2 was significantly reduced in HeLa cells treated with bcl-2 ASODN, bax and radiotherapy. ASODN and bax can induce the apopotosis cells effectively. Conclusion： Combined genes transfection of bcl-2 significantly, and enhance radiosensitivity to HeLa ceils effectively.