Study Of Enhancing Radiosensitivity Of Cervical Cancer Cells By Gold Nanoparticle

[Objective] To investigate the effect of gold nanoparticles(AuNPs)on the radiosensitization effect of cervical squamous cell carcinoma cell line C33A: preliminary study on the role of targeted radiotherapy sensitization and the effects on cell cycle and apoptosis.[METHODS]:1.AuNPs having a particle size of about 20 nm were prepared by trisodium citrate reduction tetrachloroauric acid method.2.Physicochemical properties such as particle size,distribution,and presence or absence of AuNPs were detected by dynamic light scattering(DLS),transmission electron microscopy(TEM),field emission scanning electron microscopy(FESEM),and ultraviolet spectrophotometer(UVVis)..The recovery of AuNPs prepared by trisodium citrate reduction method was determined by inductively coupled plasma mass spectrometry(ICP-AES).3.Human cervical squamous cell carcinoma cell line C33 A was selected as the experimental subject of this experiment for in vitro study.3.1 The effect of different concentrations of AuNPs on the survival rate of cell line C33 A was detected by MTT assay.3.2 C33 A cells were cultured in different concentrations of serum-free AuNPs medium.The fluorescence deposition microscope was used to observe the deposition of gold nanoparticles in cells.The appropriate concentration was used to culture cervical squamous cell carcinoma cell line C33 A.The different concentrations of AuNPs selected in the experiment were 1nM,2nM,4nM,8nM,and 16 nM,respectively.3.3 Annexin-V/PI flow cytometry was used to detect cell cycle and cell apoptosis.The experiment was divided into 4 groups,control group,simple AuNPs incubation group,simple irradiation group and AuNPs+ irradiation group.Apoptotic flow pattern was analyzed after irradiating cells with different radiation doses(2Gy,4Gy,8Gy,16Gy).[RESULTS]:1.Spherical gold nanoparticles with a particle size of about 20 nm,uniform distribution,good concentration,no obvious aggregation,and an absorption peak of about 520 and a good peak shape were successfully prepared.The average recovery was 90.94%.2.Cervical squamous cell carcinoma cell line C33 A was cultured in different concentrations of serum-free AuNPs medium.The cell viability of MTT assay was 99.4±0.45%,97.8±1.24%,98.3±0.75%,97.9±1.29%,97.5±1.058%,cervical scale.There was no significant difference in the survival rate of cancer cell line C33 A.AuNPs had no obvious cytotoxicity to cell line C33 A.According to the results of the fluorescence inverted microscope,it was confirmed that the cervical cancer cell line C33 A was cultured in a serum-free AuNPs medium having a final concentration of 4 nM for subsequent experiments.3.Cervical squamous cell carcinoma cell line C33 A was incubated in a serum-free medium at a final concentration of 4 nM.After PI staining,the cell cycle results of flow cytometry showed that the cells in G0/G1 phase were 64.40±0.58% in the control group and 58.40±0.65% in the gold nanogroup;the cells in the S phase were 21.87±0.32% in the control group and the gold nanogroup The ratio was 20.04±0.48%;the cells in the G2/M phase were11.89±0.27% in the control group and 20.43±0.76% in the gold nanogroup.The ratio of G2/M phase cells in the radiation sensitive period was significantly increased.4.Different doses of irradiation(2Gy,4Gy,8Gy,16Gy)irradiated C33 A cells,and the apoptosis rate of the blank group and the control group did not change significantly.Comparing the X-ray group with the AuNPs+X line group,the AuNPs particles can significantly enhance the effect of radiotherapy under different radiation doses.The sensitization effect of radiotherapy in the 4Gy group and the 8Gy group was the most significant,and the radiosensitization effect in the 16 Gy group was slightly reduced.[CONCLUSIONS]: 1.The gold nanoparticles were successfully prepared in this experiment,evenly distributed,no obvious aggregation,and high recovery rate.2.Gold nanoparticles have no obvious cytotoxicity to cervical squamous cell carcinoma cell line C33 A.3.Gold nanoparticles can adjust the cell cycle and promote the efficacy of radiotherapy.