Effects Of Astragalus Polysaccharide On The Growth Of Cervical Carcinoma C-4I Cells

Background and ObjectivesOne of the important biological characteristics of malignant tumor is the uncontrolled growth of cells, and the curative effect of patients with cervical carcinoma is limited by the own defects of chemotherapy and surgery. Therefore, it is of has important practical significance to study the feasibility of clinical application of Chinese medicine in the treatment of cervical carcinoma. Astragalus is a common nourishing Chinese medicinal plant. It was discovered that Astragalus Polysaccharides (APS) is the major active ingredient of Astragalus, More and more studies show that it inhibits the growth of tumor cells in leukemia, gastric cancer, and breast cancer. However, the inhibitory effects of Astragalus Polysaccharide on the growth of cervical carcinoma C-4I cells have not been reported yet. The purpose of this study is to investigate the inhibitory effects of Astragalus Polysaccharide on the growth of cervical carcinoma C-4I cells and its mechanism, so as to provide scientific basis for the feasibility of APS used in the treatment of cervical carcinoma.Methods1. APS was prepared, then it was quantitated.2. The growth rate and doubling time of cervical carcinoma C-4I cells before and after treated with APS were detected by cell growth curve.3. The change of clone-forming efficiency and the distribution of cell cycle of cervical carcinoma C-4I cells before and after treated with APS were analyzed by flow cytometry and flat clone-forming assay, respectively.4. The protein expressions of growth-related genes (cyclin A, cyclin B, cyclin E and p21) in cervical carcinoma C-4I cells before and after treated with APS were investigated by Western blotting assay.Results1. The quantitative result of prepared APS solution was 61.56mg/mL2. The results of cell growth curve showed that the growth rates of cervical carcinoma C-4I cells treated with 100μg/mL,200μg/mL and 400μg/mL of APS were significantly slowed than that in cervical carcinoma C-4I cells untreated with APS (P<0.01), and the doubling times of cervical carcinoma C-4I cells treated with 100μg/mL,200μg/mL and 400μg/mL of APS were significantly prolonged than that in cervical carcinoma C-4I cells untreated with APS (P<0.01).Combined with cells state, the cervical carcinoma C-4I cells treated with 200μg/mL APS for 48h were used as the experimental group, while the cervical carcinoma C-4I cells untreated with APS were taken as control group in subsequent experiments.3. The results of flat clone-forming ability assay showed that the clone-forming number of the cervical carcinoma C-4I cells treated with 200μg/mL APS for 48h in experimental group was significantly lower than that cervical carcinoma C-4I cells untreated with APS in control group (P<0.01), and compared with control group, the volume of the cervical carcinoma C-4I clone was smaller in experimental group.4. The data from flow cytometry suggested that compared with cervical carcinoma C-4I cells untreated with APS in control group, the G1 phase was blocked (P<0.01), and the proportion of S phase (P<0.01)and G2/M phase cells (P<0.01)were decreased obviously in cervical carcinoma C-4I cells treated with 200μg/mL APS for 48h in experimental group.5. The data from Western blotting displayed that the expressions of cyclin B and cyclin E in cervical carcinoma C-4I cells treated with 200μg/mL APS for 48h in experimental group were significantly lower than that in cervical carcinoma C-4I cells untreated with APS in control group (P<0.01), and the expression of p21 was significantly increased in cervical carcinoma C-4I cells of experimental group compared with control group (P<0.01), while the expression of cyclin A was no significant difference between experimental group and control group (P>0.05).ConclusionsAPS can significantly inhibit the growth of cervical carcinoma C-4I cells. The down-regulation of cyclin B and cyclin E expressions and up-regulation of p21 expression expression may be involved in the inhibition of APS on the growth of cervical carcinoma C-4I cells...