| Background and ObjectiveThe form and progression of tumor is a complicated and moderately advanced process, which involve neoplastic features of its own,host microenvironment and genovariation and so on. The special microenvironment of host which can adjust the progress,invasion and metastasis of tumor cell by means of diverse regulating mechanism is the soil for the tumor cell to survive and proliferation.From Rudolf Virchow found white cells in tumor microenvironment in 1863,the concrete effect on progression of inflammatory reaction in tumor microenviroment came to focus under study in the word. The agreement in the past insisted that the major effect of inflammatory reaction is to anti-tumor by participating in the immune response, compared with the present agreement that inflammatory reaction acts as a double-edged sword;it can inhibit tumor,but what is more important is that it can promote the growth and metastasis of neoplasm by various paths. In the process of inflammation promote the growth and metastasis of neoplasm, various kinds of inflammatory cells and various cytokine secreted by inflammatory cells play a important role,such as macrophages,TNF,IL-1,IL-6 and so on.We found from previous experiments that after inoculation to C57/BL6 mice, Lewis lung carcinoma cells presented with increasing growth speed, metastatic distance in the process of serial passage in mice,the phenomenon is similar to the process of tumor advanced in human.Some reports stated the similar phenomenon, and they insisted that this kind of phenomena appeared as a result of clonal selection of tumor cells in mouse. According to this theory, if the tumor cells, which have enhanced malignant potency after clonal selection by the mean of serial passage in mice, were purified and inoculated to the mices again,the growth speed and metastatic ability of the tumor cells should be enhanced. We ever primarily cultured the tumor tissue which presented with advanced ability of growth speed and metastasis after serial passage in vivo, further more, cultured in vitro many generations to remove substance cells , then inoculated the purified LLC into C57/BL6, as a result, the growth and metastasis ability of LLC recovered to the low level, whereas, the growth and metastasis ability advanced again with more serial passage of the bearing tumor mice; so we presumed that in process of this transformation, the variation of host microenvironment contributed more,compare to the tumor cellular heterogenization . Therefore, on basis of this theory, we used serial passage of the bearing tumor mice to simulate tumor cellular transformation in vivo, further to observe transformation of biological behavior as well as inflammatory reaction in microenvironment, meanwhile, to initially detect the effect of inflammation on transformation of biological behavior.Medthods1 .Cultured the LLC in complete medium, then the thrived cells were trypsinizated, 0.2ml of suspension(5×10~6/L) was inoculated subcutaneously to right armpit to establish first generation murine bearing tumor; which were killed 14days later, removed tumor and made into cell suspension(5×10~6/L), 0.2ml of which was inoculated subcutaneously to right armpit to establish second generation murine bearing tumor, repeated every 2weeks; 15 mices respectively received inoculation on the second, eighth and fifth generation, accordingly called early group, mid-half group and later group. Tumor feeling within 2-9days means model establishment done.2. Initiating tumor feeling from second day after inoculation, and wrote down the date of every group; killed all the mice in 28th day after inoculation, and observed the information of lung metastasis.3. The microvessel density( MVD), macrophages(stained by CD68) and Nuclear Factor-kappaB( NF-κB) was detected by SP test, expression of Matrixmetalloproteinase-9 (MMP-9) was detected by PV test.4.The experimentaldata was analyzed by SPSS 13.0. we also used variance analysis, x~2 testand non-parameter test; correlation analysis was done by Pearson and Spearman, P<0.05 was taken as test criterion.Result1 .With more serial passage of the bearing tumor mice, the times of oncogenesis speeds up , tumor feeling time of the early group, mid-half group and later group was day of (4.60±1.18), (3.73±1.03) and (2.93±0.96), respectively; there was significantly statistical difference (P<0.05) .2.With more serial passage of the bearing tumor mice, the rate of lung metastasis increased, the diameter and numbers of metastatic nodus increased, the lung metastasis rate of the early group, mid-half group and later group was 13.3%, 60.0%, 100%, respectively, there was significantly statistical difference (P<0.05) .3.With more serial passage of the bearing tumor mice, the macrophage infiltrated in tumortissues increased, the density of the macrophage infiltrated in tumor tissues of the earlygroup, mid-half group and later group was (24.87±8.54), (36.33±10.02),(47.20±13.49)/HP(×400), respectively, there was significantly statistical difference(P<0.05) .4.With more serial passage of the bearing tumor mice,the MVD in tumor tissue increasedsignificantly (P<0.05), the MVD of the early group, mid-half group and later group was (24.73±10.38), (41.27±13.50), (55.80±21.31)/HP(×200), respectively, and diffused in early stage turned to closed in later stage. And the increase of MVD was positively correlated with the macrophages infiltrated in tumor tissues.5.With more serial passage of the bearing tumor mice, the stain of NF-κB in tumor tissues turned stronger positive, and the positive cells also increased, the high expression rate of the early group, mid-half group and later group was 20%, 60.0%, 86.7%, respectively; MMP-9 mainly expressed in inflammatory cells and tumor cells, with more serial passage of the bearing tumor mice,the stain of MMP-9 turned stronger positive, and the positive cells also increased, but also expression of NF-κB and MMP-9 were positively correlated with density of macrophage infiltrated in tumor tissues.Conclusions1 .With more serial passage of the bearing tumor mice, the inflammatory feature in tumormicroenvironment increased.2.With more serial passage of the bearing tumor mice, the ability of tumor growth andmetastasis distance increased to follow the inflammatory feature in tumormicroenvironment increased.3. inflammatory reaction in microenvironment contributes to progress of tumor. |
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