| Objective: Gastric cancer is one of the most common malignant tumors. Due to the low rate of early diagnosis, most patients in the clinic has been in progress. Currently chemotherapy has become the main treatment for advanced gastric cancer and recurrence of gastric cancer,some paitients can’t tolerate chemotherapy.However traditional Chinese Medicine(TMC)is not only low toxicity, and can improve the body’s immune system. TMC combined with chemotherapy for gastric cancer has been gradually applied in clinical. This research will study the tumor from gastric cancer subcutaneous transplanted nude mice established by BGC823 cells. Observe the influence of curcumol and fluorouracil on tumor proliferation,and explore the mechanism of TMC treatment of gastric cancer.Methods:1 Establishiment of nude mice subcutaneous transplantation model. Collect human gastric BGC823 cells in fine growth state cell were inoculated subcutaneously in nude mice axilla. The subcutaneous implantatio tumor model of human gastric cancer in nude mice was built.2 When the subcutaneous tumor growth to 100 mm, The nude mice were randomly divided into 4group,the control group, the curcumol group, the fluorouracil group and the curcumol combined with fluorouracil group. The control group received saline; the curcumol group received the curcumol; the fluorouracil group received fluorouracil; the curcumol combined with fluorouracil group received the curcumol and fluorouracil.Every 4 days to give the drug intratumoral injection, the mouse were sacrificed after 4 days of the last drug intervention.3 Detection the volume and weight of human gastric cancer BGC823 cell nude mice subcutaneous transplantation tumor.Measure the longest diameter(A)and the shortest diameter(B)of subcutaneous transplantation tumor before each drug intervention and after sacrifice nude mouse.According to the formula,the volume(mm3)of the tumor=1/2×A×B2.Theapproximate volume of the subcutaneous transplantation tumor was calculated.After sacrificed the nude mouse,the whole of the tumor was takenout and measured.The inhibition rate of the subcutaneous transplantation tumor=(the tumor weight of the control group-the tumor weight of theexperimental group)/the tumor weight of the control group×100%.4 Proliferation of the human gastric cancer BGC823 cell was detected in 0,24,48,72 hours by MTT.Four group of different facters treated tumor were configured to cell suspension.The OD values of each group on 0,24,48,72 hours were measured and the inhibition rate of proliferation of the tumor were calculation.5 Detect the expression of PCNAm RNA,P27 m RNA in four groups of tumor by q RT-PCR.6 Detect the expression of PCNA and P27 protein in four groups of tumor by immunohistochemistry.Results: 1 The model of nude mice subcutaneous transplantation tumor was built.The model of nude mice subcutaneous transplantation tumor was successfully establishment.The subcutaneous transplantation tumor formation rate was 100%(20/20). 2 The effect of curcumol and fluorouracil on the subcutaneous transplantation tumor of human gastric cancer BGC823 cell in nude mice 2.1 The volume of the subcutaneous transplantation tumorThe final volume of the tumor in the control group(2195.83±54.00) mm3 was significantly bigger than that in the curcumol group(1795.48±56.86) mm3, the fluorouracil group(1439.50±31.13) mm3 and the curcumolcombined with fluorouracil group(1363±37.54) mm3,the difference was statistically signi- ficant(P<0.05).Compared with the curcumol group,the inhibition rate of the volume of the tumor in the curcumol+5-FU group was significantly higher(P<0.05).But compared with the fluorouracil group,the final volume of the tumor in the curcumol combined with fluorouracil group was no significant difference(P>0.05). 2.2 The inhibition rate of the subcutaneous transplantation tumorThe tumor weight in the control group(1.88±0.09)g was significantly more than that in the curcumol group(1.51±0.99) g,in the fluorouracil group(1.27±0.02)g, in the curcumol combined with fluorouracil group(1.05±0.10)g and they were significant difference(P<0.05). Compared with the curcumol group, the inhibition rate of the weight of the tumor in the curcumol combined with fluorouracil group was significantly higher(P<0.05). But compared with the fluorouracil group, the weight of the tumor in the curcumol combined with fluorouracil group was no significant difference(P>0.05). 3 The effect on the proliferation of the subcutaneous transplantation tumor of human gastric cancer cell in nude mouse.MTT results showed that:the OD values of the control group, the curcumol group, the fluorouracil group and the curcumol combined with fluorouracil group in 0 hours were 0.129±0.023,0.131±0.019, 0.126±0.020, 0.128±0.021 respectively;the OD values in 24 hours were 0.236±0.054, 0.232±0.041,0.238±0.037,0.234±0.050 respectively; the OD values in 48 hours were 0.512±0.056,0.416±0.068,0.382±0.069,0.358±0.067 respectively; the OD values in 72 hours were 0.702±0.051,0.623±0.058, 0.562±0.061, 0.519±0.082.There are no obvious difference in the group of 0 hours and 24 hours(P > 0.05).But experimental groups were significantly less than the control group in 48 hours and 72 hours(P<0.05). 4 The expression of the PCNAm RNA,P27 m RNA of nude mice subcutaneous transplantation tumor tissues 4.1 The expression of the PCNA m RNAThe expression of PCNA m RNA of tumor tissues in the control group(1.001±0.0682) were significantly higher than that in the curcumol group(0.8673±0.0452), the fluorouracil group(0.07553±0.0460) and the curcumol combined with fluorouracil group(0.5327±0.0284),the difference werestatistically significant(P<0.05). Compared with the curcumol group and the fluorouracil group, the expression of PCNA m RNA of tumor tissues in the curcumol combined with fluorouracil group was significantly lower(P<0.05). 4.2 The expression of the P27 m RNAThe expression of P27 m RNA of tumor tissues in the control group(1.010±0.1746) were significantly lower than that in the curcumol group(1.8973±0.3752), the fluorouracil group(2.0274±0.2319) and the curcumol combined with fluorouracil group(2.5566±0.3268),the difference were statistically significant(P<0.05). Compared with the curcumol group, the expression of P27 m RNA of tumor tissues in the curcumol combined with fluorouracil group was significantly higher(P<0.05). But compared with the fluorouracil group, the expression of P27 m RNA of tumor tissues in the curcumol combined with fluorouracil group was no significant difference(P>0.05). 5 The expression of PCNA protein and P27 protein in the nude mice subcutaneous transplantation tumor cells of BGC823 by curcumol and fluorouracil. 5.1 The positive percentages of PCNA protein in the control group, the curcumol group, the fluorouracil group and the curcumol combined with fluorouracil group were respectively(73.56±2.26)%,(56.94±2.58)%,(44.05±1.70)%,(40.17±2.90)%.The positive percentages of PCNA protein in the experimental groups were significantly lower than the control group, the difference were statistically significant(P<0.05). But compared with the fluorouracil group, the positive percentages of PCNA protein in the the curcumol combined with fluorouracil group was no significant difference(P>0.05). 5.2 The positive percentages of P27 protein in the control group, the curcumol group, the fluorouracil group and the curcumol combined with fluorouracil group were respectively(33.67±2.36)%,(51.06±1.98)%,(60.28±2.01)%,(71.67±2.29)%.The positive percentages of PCNA protein in the experimental groups were significantly higher than the control group, the difference were statistically significant(P<0.05). Compared with the curcumol group and the fluorouracil group, the positive percentages of PCNA protein in the curcumol combined with fluorouracil group was significantly lower(P<0.05).Conclusions:1 Curcumol can inhibit the growth of human gastric cancer BGC823 cell nude mice subcutaneous transplantation tumor.2 Curcumol can reduce the growth of cells of human gastric cancer BGC823 cell nude mice subcutaneous transplantation tumor.3 Curcumol combined with fluorouracil can inhibit the growth of human gastric cancer BGC823 cell nude mice subcutaneous transplantation tumor, compared with the curcumol, the difference were statistically significant.4 Curcumol combined with fluorouracil can inhibit the growth of human gastric cancer BGC823 cell nude mice subcutaneous transplantation tumor, but with the single fluorouracil was no statistically significant differences.5 Curcumol can inhibit the growth of subcutaneous transplantation tumor. The mechanism may be reduced expression of PCNA,P27 upregulation. |
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