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Objective: To study the radiosensitivity of SiHa cells with Ku80 inhibition by RNAi. Methods: pKu80-siRNA and pNeg-siRNA were constructed and then transfected into SiHa cells. Western blot and RT-PCR were applied to measure the expression of Ku80. After 10 Gy irradiation with 6 MV X-ray, cells were harvested at 20 h, 28 h and 48 h, and analyzed by flow cytometry for apoptosis rate. The SF2 and α, β values of cells were acquired by clone formation array. Results: Two stable transfection cell clones SiHa/Ku80-siRNA and SiHa/Neg-siRNA were screened from the pKu80-siRNA and pNeg-siRNA transfected cells. Western blot and RT-PCR indicated that the expression of Ku80 was suppressed by the Ku80-siRNA. The apoptosis rates of SiHa/Ku80-siRNA were higher than control cells at 28 h and 48 h after X-ray irradiation (P < 0.05). The SF2 value of SiHa/Ku80-siRNA (0.422) was lower than that of SiHa/Neg-siRNA cells (0.587) or untransfected cells (0.583), and the α value of SiHa/Ku80-siRNA was 0.295 ± 0.016, higher than that of SiHa/Neg-siRNA (0.176 ± 0.010) or control cells (0.188 ± 0.011). Conclusion: Inhibition of Ku80 could enhance the radiosensitivity of SiHa, which showed that Ku80 could be a good target for molecular therapy.
Objective: To study the radiosensitivity of SiHa cells with Ku80 inhibition by RNAi. Methods: pKu80-siRNA and pNeg-siRNA were constructed and then transfected into SiHa cells. Western blot and RT-PCR were applied to measure the expression of Ku80. After 10 Gy irradiation with 6 MV X-ray, cells were harvested at 20 h, 28 h and 48 h, and analyzed by flow cytometry for apoptosis rate. The SF2 and α, β values of cells were acquired by clone formation array. Results: Two Western blot and RT-PCR showed that the expression of Ku80 was suppressed by the Ku80-siRNA. The results showed that the expression of Ku80 was inhibited by the Ku80-siRNA. The SF2 value of SiHa / Ku80-siRNA (0.422) was lower than that of SiHa / Neg (P <0.05). The apoptosis rates of SiHa / Ku80- siRNA were higher than that of control cells at 28 h and 48 h after X-ray irradiation -siRNA cells (0.587) or untransfected cells (0.583), and the α value of SiHa / Ku80-siRNA was 0.295 0.016, higher than that of SiHa / Neg-siRNA (0.176 ± 0.010) or control cells (0.188 ± 0.011) Conclusion:. Inhibition of Ku80 could enhance the radiosensitivity of SiHa, which showed that Ku80 could be a good target for molecular therapy.