Dissertation
Dissertation > Medicine, health > Oncology > Genitourinary tumors > Female genital tumors > Uterine tumors

The Study on the Radiosensitizing Effect and Mechanism of Artemisinin on Nude Mouse Transplanted with HeLa and Siha Cells of Human Cervical Cancer

Author FengYang
Tutor CaoJianPing
School Suzhou University
Course Radiation Medicine
Keywords Artemisinin Radiosensitising Transplantation tumor Human cervical cell Microvessel density Apoptosis Cell cycle Cycle regulatory protein
CLC R737.33
Type Master's thesis
Year 2011
Downloads 94
Quotes 0
Download Dissertation

Objective To investigate the effect of artemisinin on radiosensitivity of nude mice transplanted with p53 mutant type HeLa cells and p53 wild type Siha cells of human cervical cancer. And try to find out the mechanism of radiosensitivity by comparing and analyzing the difference of radiosensitising effect of art on mice transplanted with different p53 states of human cervical cancer cells. Methods The experiment was based on 20 nude mice with subcutaneoustransplantation tumor of two different p53 states of human cervical carcinoma. They were divided randomly into four groups:control group; artemisinin group; simplicity radiation group and radiation plus artemisinin group. Control group and simplicity radiation group were injected intraperitoneal with 0.2 ml/d sterile seed fat, radiation plus artemisinin group and artemisinin group were injected intraperitoneal with 50 mg.kg-1.d-1 sterile seed fat mixed with artemisinin, each group treated for 6 days. Locally radiated nude mice transplanted tumor at a dose of 10 Gy after 6 days. 14 days after radiation nude mice were sacrificed.(1) The tumor size and weight were measured, calculate tumor inhibitory rate and growth inhibition rate; (2) The changes of microvessel density and apoptosis rate of transplantation tumor distribution effected by artemisinin combined withγ-ray irradiation was detected by TUNEL and immunohistochemistry technique;(3) The changes of cell cycle of transplantation tumor distribution effected by artemisinin combined withγ-ray irradiation was detected by flow cytometry;(4) Changes of expression of cell cycle regulatory protein wee1、cyclinB1、cdc2 of transplantation tumor that caused by artemisinin combined withγ-ray irradiation was investigated by Western-blot assay.Results (1) Transplantation tumor of p53 mutant type HeLa cells volume and weight of radiation plus artemisinin group were significantly different from those of other three groups ( P<0.05 ),but there was no difference between artemisinin group and control group(P>0.05). In transplantation tumor of Siha cells the volume and weight in radiation plus artemisinin group exhibted no difference with radiation group(P>0.05), and there was no difference between artemisinin group and control group(P>0.05). The inhibitory rate and growth inhibition rate of transplantation tumor of p53 mutant type HeLa cells in the artemisinin group, radiation group and radiation plus artemisinin group is 4.00%, 31.49%, 58.03% and 5.57%, 29.02%, 51.03%;In Siha cells,the inhibitory rate and growth inhibition rate is 0, 33.00%, 38.13% and 0, 37.39%, 28.94%. (2) The assays of microvessel density showed:transplantation tumor of HeLa cells of radiation plus artemisinin group was different from that of other three groups ( P<0.05 ),but there was no difference between artemisinin group and control group(P>0.05);transplantation tumor of Siha cells in radiation plus artemisinin group had no difference with radiation group(P>0.05), and there was no difference between artemisinin group and control group(P>0.05).(3)TUNEL assays showed: The apoptpsis induction of transplantation tumor of HeLa cells in radiation plus artemisinin group had difference with other three groups ( P<0.05 ),but there was no difference between artemisinin group and control group(P>0.05). In Siha cells,the apoptpsis induction in radiation plus artemisinin group had no difference with that of radiation group(P>0.05), and there was no difference between artemisinin group and control group(P>0.05). (4) Flow cytometry analysis showed: compared with simplicity radiation group,the percent age of cells of radiation plus artemisinin group in G0/G1 phase increased (P<0.05) and G2/M phase decreased (P<0.01) in transplantation tumor of HeLa cells. In transplantation tumor of Siha cells,the percent age of G0/G1 phase and G2/M phase didn’t show obvious change (P>0.05) in the groups of radiation combined with or without artemisinin. (5) Western blot analysis showed:in transplantation tumor of HeLa cells, there was a wee1 decreased, cyclin B1 increased expression and cdc2 had no significant change in radiation plus artemisinin group. In transplantation tumor of p53 wild type Siha cells, expression level of wee1, cyclin B1 and cdc2 had no significant change in all groups.Conclusions (1)Artemisinin can not inhibit the growth of transplantation tumor in the nude mice, but artemisinin enhanced can inhibit the growth of transplantation tumor of p53 mutant type HeLa cells. However had no such effect on transplantation tumor of p53 wild type Siha cells;(2) Artemisinin combined withγ-ray irradiation can decrease the microvessel density of transplantation tumor of HeLa cells,but exhibit no such effect on transplantation tumor of Siha cells;(3) Artemisinin can increase the apoptosis of transplantation tumor of HeLa cells which were irradiated,but shows no such effect on transplantation tumor of Siha cells;(4) Artemisinin can decrease the cell G2/M arrest of transplantation tumor of HeLa cells after irradiation; (5)The exposure ofγ-ray can cause a G2/M arrest in transplantation tumor of HeLa cells corresponding to an increased expression level of wee1 and an decreased expression of cyclin B1. However, the cells of transplantation tumor of Siha were arrested in G1/S phase after irradiation, so the expression of protein that related to G2/M phase had no changes in response to irradiation;The expression of cdc2 exhibits no significant change in both transplantation tumor of HeLa and Siha cells after therapy of drug combination with radiation .

Related Dissertations
More Dissertations