| 1. Studies on the methodology of 18F-FDG cell binding of breast cancer cell Bcap37Objective: To establish a useful and stable methodology of 18F-FDG cell binding of breast cancer cell Bcap37.Methods: Several different binding conditions were tested. (1)The cell number was 1.25×105,2.5×105,5×105,1×106,5×106 and 1×107 /bottle. (2)The reaction time was 20,40,60,80,100 and 120min. (3)The radioactivity of 18F-FDG was 1.85,3.7,7.4,14.8 and 29.6KBq. (4)The glucose concentration was 0,1.39,2.78 and 5.5mmol/L. The count of CPM(B) in cells and CPM(F) in supernatant were measured by theγequipment .The binding efficiency of breast cancer cell line Bcap37 with 18F-FDG was calculated.Result: (1)When other conditions were unchanged①The binding efficiency was from(6.00±1.46)%,(13.14±1.32)%,(16.74±2.11)%,(33.82±3.70)%,(36.03±4.33)% and (37.40±4.23)% when the cell number was changed from 1.25×105 to 1×107 /bottle. The binding efficiency of breast cancer cell increased with the increase of cell number. The difference of binding efficiency was no significant between 2.5×105 and 5×105 group. The difference of binding efficiency was no significant between 1×106, 5×106 and 1×107 group. The difference of binding efficiency was significant between the others (F=75.895,P<0.05).②The binding efficiency was(14.78±3.26)%,(21.86±3.25)%,(24.14±2.30)%,(27.03±3,97)%,(34.09±1.96)% and(35.00±1.85)% when reaction time was from 20min to 120min. The difference of binding efficiency was no significant between 40min, 60min and 80min group. The difference of binding efficiency was no significant between 100min and 120min group. The difference of binding efficiency of the remain of groups were significant ( F=35.594,P<0.05).③The binding efficiency was (38.09±5.81)%, (38.07±1.36)%, (38.84±3.54)%, (35.81±1.21)% and (36.88±2.79)% when radioactivity of 18F-FDG was from 1.85 to 29.6KBq. The difference of binding efficiency was no significant in all groups(F=3.477,P>0.05).④The binding efficiency was (35.41±1.95)%, (25.93±2.17)%, (15.40±5.53)%, (2.38±0.93)% when glucose concentration was from 0 to 5.5mmol/L. The difference of binding efficiency was significant in the all groups (F=100.739,P<0.05).(2)The basic conditions of 18F-FDG cell binding of breast cancer Bcap37: the cell number was 1×106/bottle, the radioactivity of 18F-FDG was 3.7KBq, the glucose concentration was 0mmol/L, the reaction time was 100min, the binding efficiency was (34.09±1.96)%.Conclusions: The methodology of 18F-FDG cell binding of breast cancer Bcap37 was established successfully. It contributed to evaluate the effect of chemotherapy on inhibiting breast cancer cell line Bcap37 and to compare with a drug-resistant cell Bcap37/MDR1.2. Comparative Studies of the18F-FDG cell binding the drug-resistan breast cancer cell Bcap37/MDR1 and breast cancer cell Bcap37Objective: to compare the18F-FDG cell binding in the drug-resistan breast cancer cell Bcap37/MDR1 and breast cancer cell Bcap37Methods: Several different binding conditions were tested. (1)The cell number was 1.25×105,2.5×105,5×105,1×106,5×106 and 1×107 /bottle.(2)The reaction time was 20,40,60,80,100 and 120min. (3)The radioactivity of 18F-FDG was 1.85,3.7,7.4,14.8 and 29.6KBq. (4)The glucose concentration was 0,1.39,2.78 and 5.5mmol/L.The count of CPM(B) in cells and CPM(F) in supernatant were measured by theγ equipment .The binding efficiency of breast cancer cell line Bcap37/MDR1 with 18F-FDG was calculated.Result: (1)When other conditions were unchanged①when the cell number was changed from 1.25×105 to 1×107/bottle,the binding efficiency of Bcap37 was (6.00±1.46)%, (13.14±1.32)%, (16.74±2.11)%, (33.82±3.70)%, (36.03±4.33)% and (37.40±4.23)% and the binding efficiency of Bcap37/MDR1 was (2.24±0.7)%, (5.00±0.63)%, (11.58±1.62)%, (20.74±2.19)%, (24.41±2.31)% and (28.98±1.95)% . The binding efficiency of Bcap37 was higher than the Bcap37/MDR1, the difference was significant (P <0.05).②When the reaction time was 20,40,60,80,100 and 120min, the the binding efficiency of Bcap37 was (14.78±3.26)%, (21.86±3.25)%, (24.14±2.30)% , (27.03±3.97)%, (34.09±1.96)% and (35.00±1.85)% and the binding efficiency of Bcap37/MDR1 was (6.35±1.87)%, (12.18±0.42)%, (15.92±2.19)%, (18.59±1.29 )%, (21.73±1.27)% and (23.32±1.37)%. The binding efficiency of Bcap37 was higher than the Bcap37/MDR1, the difference was significant (P <0.05).③when the 18F-FDG radioactivity were 1.85,3.7,7.4,14.8 and 29.6KBq, the binding efficiency of Bcap37 was (38.09±5.81)%, (38.07±1.36)%, (38.84±3.54)%, (35.81±1.21)% and (36.88±2.79)% and the binding efficiency of Bcap37/MDR1 was (18.98±0.76)%, (19.53±1.11)%, (19.56±0.76)%, (19.7±0.7)% and (19.27±0.68)%. The binding efficiency of Bcap37 was higher than the Bcap37/MDR1, the difference was significant (P <0.05).④when the concentration of glucose 0,1.39,2.78 and 5.5mmol / L, the binding efficiency of Bcap37 was (35.41±1.95)%, (25.93±2.17)%, (15.40±5.53)% and ( 2.38±0.93)% and the binding efficiency of Bcap37/MDR1 was (19.82±2)%, (14.97±1.88)%, (11.90±1.51)% and (2.07±0.77)%. The difference of binding efficiency on Bcap37 and Bcap37/MDR1 cell at the 0 and 1.39 mmol/L groups was significant (P <0.05); The difference of binding efficiency on Bcap37 and Bcap37/MDR1 cell at 2.78 and 5.5 mmol/L groups was not statistically significance (P> 0.05). (2)The basic conditions of 18F-FDG cell binding of breast cancer Bcap37: the cell number was 1×106/bottle, the radioactivity of 18F-FDG was 3.7KBq, the glucose concentration was 0 mmol / L, the reaction time was 100min. the binding efficiency was (19.53±1.11)%.Conclusions: The methodology of 18F-FDG cell binding of breast cancer Bcap37/MDR1 was established successfully. The best conditions of 18F-FDG cell binding of breast cancer Bcap37/MDR1 was same with the Bcap37, but the 18F-FDG cell binding of breast cancer Bcap37/MDR1 was lower than the Bcap37. It Shows that the proliferation and metabolism of resistance of tumor cell are lower than the parental tumor cells .It contributed to use the 18F-FDG cell binding to evaluate the effect of chemotherapy on inhibiting breast cancer cell line Bcap37/MDR1and Bcap37.3. Experimental studies of 18F-FDG cell binding valuating the inhibitory effect of chemotherapy in human breast cancer cell line Bcap37andBcap37/MDR1Objective: To compare the different of early valuating the inhibitory effect of chemotherapy in breast cancer cell Bcap37 and Bcap37/MDR1 with 18F-FDG cell binding experiment and MTT.Methods: (1) To draw up cell growth curve according to the optical density. (2) MTT was ultilized to measure the inhibitory rate on Bcap37 and Bcap37/MDR1 at different concertrations of Adriamycin and Gemcitabine at 48 hours.(3) Binding experiment was utilized to measure the inhibitory rate on Bcap37 and Bcap37/MDR1 at different concentrations of Adriamycin and Gemcitabine at 24 hours.Result: (1) With the method of MTT, the difference of the inhibitory rate on Bcap37 and Bcap37/MDR1 cell at every same concentration of Adriamycin was significant (P<0.05). The difference of the inhibitory rate on Bcap37 and Bcap37/MDR1 cell at every same concentration of Gemcitabine was no significant(P>0.05).(2) With the method of 18F-FDG cell binding, the difference of the inhibitory rate on Bcap37 and Bcap37/MDR1 cell at same concentrations of Adriamycin was significant(P<0.05). The difference of the inhibitory rate on Bcap37 and Bcap37/MDR1 cell at same concentrations of Gemcitabine was not significant(P>0.05).Conclusions: Bcap37 cell was sensitive to Adriamycin and Gemcitabine; Bcap37/MDR1 cell was sensitive to Gemcitabine and insensitive to Adriamycin. 18F-FDG cell binding experiments could be used for early measurements and evaluation of the inhibitory effection of Adriamycin and Gemcitabine on breast cancer. It is one of the indicators of early evaluate effection of chemotherapy and the resistance of tumor cell . |
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