| Background:Gastric cancer is a malignant tumor which has the highest morbidity and mortality in gastrointestinal cancer in China.Though the treatments of gastric cancer have been improved,the 5-year survival rate of gastric cancer is still hovering 20% to 30% all over the world for nearly 30 years.Tumor metastasis has a great impact on postoperative survival.Through a long time of clinical practice and experimental research, Professor Wei-pinkang thinks that phlegm is an important material foundation of cancer.He has established the phlegm Syndrome theory, then devised the Xiaotan Sanje recipe, which has achieved satisfactory results.Heparanase is a newly found enzyme which can deform extra cellular matrix and basement membrane and reduce microvessel density,which plays an important role in tumor metastasis.Due to effect of heparanase we established the nude mice model of orthotopic transplantation and deeply study the possible inhibition mechanism of the metastasis of Xiaotan Sanjie Recipe.Objective:The purpose of this study is to discover the mechanism of Xiaotan Sanjie Recipe restraining the metastasis of the human gastric carcinoma implanted in nude mice model which is similar to the style of human cancer invasion and metastasis, by examining the change of CK20mRNA and MVD and heparanase.Methods:1.The screened MKN-45 cancer cell line was implanted in BALB/C nude mice stomach near the greater curvature side, nude mice transplantation tumor model of human gastric cancer was set up, the growth of tumor and the life status of mice were observed regularly.The animals were sacrificed and observed the local growth of transplanted tumor was observed.2.30 BALB/C nude mice were divided into 3 groups completely and randomly:control group,Xiaotan Sanjie Recipe group,5-Fu group.The mice in control group were given isotonic saline 0.4ml ig once a day; in herb groups given Xiaotan Sanjie Recipe 0.4ml once a day; in 5-Fu group given 5-Fu dilution (60mg.kg-1.w-1) 0.2ml ip once a week×3w.The course of treatment was 6 weeks until the end.3.After 6 weeks, CK20mRNA was examined in the blood samples collected from the eyes by the method of RT-PCR.Then the mice were sacrificed.The tumor were collected and weighted.The livers metastasis were counted before making HE staining and pathological identification.4.The expression of Heparanase in tumor tisssues was examined by means of immunohistochemical staining, and MVD counted.Results:1.We built an orthotopic implantation model of human gastric carcinoma MKN-45 in nude mice successfully.The lump could be touched in the upper abdominal afer two weeks which was hard and growing bigger gradually.The tumors were gray-white after the abdomen was opened.The formation rate was 100%. The tumor inhibition rate was 56.3%,47.2% respectively.The rates of both Xiaotan Sanjie recipe group and 5-Fu group were different significantly (P=0.000,P<0.05) compared with the controlled group.But there are no different between the experiment group (P=0.163,P>0.05).At the same time,liver metastasis of the Xiaotan Sanjie group is significantly fewer than that of the control group (P=0.07,P<0.05),and the 5-Fu group are fewer than the control group too (P=0.03,P<0.05).But there is no significant difference between that of Xiaotan Sanjie group and the 5-Fu group(P=0.538,P>0.05).2.CK20mRNA is expressed in all of the four groups.Compared with the controlled group,there is obviously different between the controlled group and the control goup (P=0.004, P<0.01),but no differentce of 5-Fu group (P=0.567, P>0.05)3.The results of immunohistochemical staining indicate that the group of Xiaotan Sanjie Recipe is statistically different compared with the controlled group (P=0.00, P< 0.05).5-Fu group has no obvious effect on the construction of Microvessel (P=0.584, P> 0.05).Microvessel density and heparanase were detected in the tumor tissue by the means of immunohistochemical staining.Compared with the controlled group,Xiaotan Sanjie Recipe group is significantly different (P=0.01,P<0.05),however there is no significance between 5-Fu group and controlled group (P=0.248, P>0.05)Conclusion:1.The orthotopic implantation model of human gastric carcinoma MKN-45 in nude mice was successfully build.Xiaotan Sanjie Recipe can obviously inhibit the growth of the tumor the orthotopic implantation model of human gastric carcinoma MKN-45 in nude mice.It can decrease the metastasis of the liver on the same time.2. The Xiaotan Sanjie Recipe can down-regulate the CK20mRNA in the peripheral blood.It can obviously inhibit the tumor cell to enter the peripheral blood and reduce the metastasis of the tumor.3. The Xiaotan Sanjie Recipe efficiently obviously reduce MVD in the tumor of the orthotopic implantation model of human gastric carcinoma MKN-45 in nude mice.4. The Xiaotan Sanjie Recipe can efficiently decrease the heparanase in the tumor the orthotopic implantation model of human gastric carcinoma MKN-45 in nude mice. |
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