The Study On Enhancing Radiosensitivity Of Hela Cells By Combined Genes Transfection Of Bcl-2ASODN And Bax.

Objective 1.To investigate the apoptotic effect of human cervical cancer cell lines by transfect antiapoptosis gene-bcl-2AS0DN and pro-apoptotic gene-bax and further explore the value of combined genes transfection on enhancing the radiosensitivity of Hela cells. 2.To investigate the inhibition effects of bcl-2 ASODN and bax combined transfection on the human cervical cancer transplanted subcutaneously in nude mice.Methods 1.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 3d. 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. 2. The Hela cells were cultured and implanted subcutaneously into 25 nude mice .At the 14th day, these mices were divided into five groups: normal control group, bcl-2SODN group, bcl-2ASODN group, bax group and bcl-2ASODN+bax group. After different treatment, the tumor volumes were measured and inhibition rate was calculated. Terminal Deoxynucleotidyl Transferase-mediated Nick End Labeling (TUNEL) was used to detect the apoptosis.Results 1.① After genes transfection and radiotherapy a significant apoptosis was observed by microscopy. ②The inhibition ratio and apoptosis of cells treated with genes transfection and radiotherapy were significantly higher than that of other groups(P<0.05). ③Compared 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-2ASODN ,bax and radiotherapy. 2.Two weeks after Hela cells implantation, all mice formed solid tumor, and the inhibition rate of ASODN group showed significant difference vs other groups at the 17th day (P< 0.01) . The inhibition rate of bcl-2 ASODN+bax group was 40.0% at the termination day, and displayed significant difference when compared with other groups (P<0.01). The number of apoptosis cells in bcl-2 ASODN+bax group was 40.5±4.9 per HPF, there was also significant difference compared with other groups (P<0.01).Conclusions 1.Combined genes transfection of bcl-2 ASODN and bax can induce the apopotosis significantly, and enhance the radiosensitivity of Hela cell lines effectively. 2.Combined genes transfection of the bcl-2ASODN and bax can more effectively inhibit cervical xenograft tumor growth than monogenic transfection via promoting the apopotosis of tumor cell significantly.