| Objective: Cervix lesion is one of the most common suffering in women, and it can develop to cervical carcinoma .In female carcinoma, the incidence of cervical carcinoma is second only to mammary cancer. In our country, cervical carcinoma is very a common malignant tumor. Usually, we think that radiotherapy and surgery are the most effective therapy, but in high risk cases, such as late phase carcinoma, coop carcinoma, lymph-node metastasis or small cell carcinoma radiotherapy and surgery are not effective. The main cause of failure is local relapse and metastasis. Even with modern instrument and radical operation, cervical carcinoma is difficult to cure. With the progress of new drug and chemo-therapy technic, chemo-therapy is more and more important. However, most anti-tumor drugs are short of selective inhibition to cervical carcinoma, and when they kill tumor cells, they also make toxicity. In this study, we investigate the effect of mifepristone and garlic oil to Hela cells, and seek to new method curing cervix carcinoma.Methods: 1 .Cell culture: Routine method revived Hela cells freezed in liguid nitrogen, and inoculated in RPMI-1640 blended with 10% foetus Cattle blood serum, cultured inculture bottle, then put the bottle in the incubator with the condition of 37 , 5%CO2. We observed the growth of cells every day. When 70% of the cells confluence, We digested them with EDTA and then made them into cell suspension. 2. MTT estimating the growth suppression of cell: We adjusted deuterium of cells to 3 104 / ml, mixed them completely and inoculated them to cultivation Plate of 96-pore, 200 l in every pore. We put them to incubator and cultured them to logarithm phrase. Then we got them out ,got rid of the superior liquor and Added 200 l culture medium including drug, the Concentration of drug are respectively 5umol/L , 10umol/L , 20umol/L , 40umol/L(mifepristone),and 12.5 mg/L, 25mg/L, 50 mg/L, 100 mg/L(garlic oil),then put them back to incubator, watching them everyday .We added 20 1 MTT in every pore after cultured 24 , 48 , 72 hours, then cultured them in incubator again for 4 hours, after this ,we got rid of all liquor ,added 200 1 di-methyl-Sulfoxide, shocking for 8 minutes,and then measured the number of A and calculate the inhibition rate .we did it for 3times in the same way.3. Observation by light microscope: We adjusted deuterium of cell to 3 104 / ml, mixed them completely and inoculated them to cultivation Plate of 24-pore with cover glass in it ,1ml in every pore, then put them to incubator and cultured them to logarithm phrase, then we got them out ,got rid of the superior liquor, and then added drug in them, wegot out cover glass after 72 hours.we Stained them with Gimsa,and observed them by light microscope.4. observation by electron microscope: We inoculated cells in the above concentration, and then added drug to them .We collected cells after 24, 48 , 72 hours , immobiled in 4% Glutaral , desiccated, soaked embedded in epikote, and make them into lipo- piece, after having been cut into ultrathin section, we observed them by transmission electron microscope.5. Assayed by TUNEL Test: We got cell sheet in the same way above, we treated them according to instruction of TUNEL Test Kit. Nucleus of cells which were stained by brown were Apoptosis cells, but the blue are negative. We counted 300 cells in every section, then calculated the rate of apoptisis cells.Result: 1. Garlic oil of four different concetrations inhibited the growth of cells after added to drug for 24, 48 , 72hours, showing the time and dose dependency. The inhibition rate were respectively 38.46%, 56.25%, 65.32%, 76.33% for 72 hours. 2. Mifepristone of four different concetrations inhibited the growth of cells after added to drug for 24, 48, 72hours, showing the time and dose dependency. The inhibition rate were respectively 36.74%, 49.62%, 61.96%, 71.34%. 3.When garluic oil was added with 0.5 g/ml cisplatin,the inhibition rate were 38.37%, 51.55%, 63.87% , 75.21%; When mifeprist...
|
PDF Full Text Request