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目的探讨转移前期原发黑色素瘤对肺组织微环境的作用及其对转移的影响。方法将黑色素瘤细胞B16种植于雌性Balb/c小鼠背部,建立荷瘤小鼠动物模型;通过小鼠尾静脉注射B16细胞建立转移模型;塞来昔布灌胃处理小鼠建立抗炎治疗模型。采用肺组织干湿比和HE染色分析肺组织炎症反应,应用ELISA试剂盒检测血清及细胞培养上清液中的细胞因子水平。结果在转移前期,与对照组小鼠相比,实验组荷瘤小鼠肺组织明显水肿,肺干湿比明显升高,两组相比差异有统计学意义(P<0.001);同时发现B16细胞易于出现在肺组织炎症细胞聚集的部位。ELISA检测结果显示,与对照组相比,实验组荷瘤小鼠血清VEGF、M-CSF和TNF-α含量明显升高,两组相比差异有统计学意义(P<0.001)。B16培养上清可以显著诱导肺组织炎症反应,与对照组相比,实验组小鼠肺组织转移瘤显著增多(P<0.001);给予抗炎药物塞来昔布处理后,与对照组相比,实验组小鼠肺组织炎症反应明显下降,肺转移瘤数目明显减少(P=0.005)。结论在肿瘤转移前期,原发恶性黑色素瘤能够调节肺组织微环境,诱导肺组织炎症反应并促进肺转移;塞来昔布可缓解肺组织炎症反应并抑制肺转移。
Objective To investigate the effect of primary melanoma on lung microenvironment and its metastasis in early stage of metastasis. Methods B16 melanoma cells were implanted into the back of female Balb / c mice to establish animal models of tumor-bearing mice. B16 cells were injected into the tail vein of mice to establish a metastatic model. The mice were treated with celecoxib and intragastrically administered to establish anti-inflammatory model . The lung tissue was analyzed for the inflammatory response by the ratio of wet and dry lung tissues and HE staining. The levels of cytokines in serum and cell culture supernatants were detected by ELISA kit. Results In the early stage of metastasis, compared with the control group mice, the lung tissue of the tumor-bearing mice in the experimental group had obvious edema and the wet / dry ratio of the lung was significantly increased. There was significant difference between the two groups (P <0.001) Cells are prone to accumulate in inflammatory cells in the lung tissue. Compared with the control group, the levels of VEGF, M-CSF and TNF-α in the tumor-bearing mice in the experimental group were significantly increased as compared with the control group, and the difference was statistically significant (P <0.001). B16 culture supernatant can significantly induce lung inflammation, compared with the control group, mice in the experimental group significantly increased lung metastases (P <0.001); given anti-inflammatory drug celecoxib treatment, compared with the control group , The inflammatory reaction of lung tissue in experimental group decreased obviously, and the number of lung metastases significantly decreased (P = 0.005). Conclusion In the early stage of tumor metastasis, primary malignant melanoma can regulate the microenvironment of lung tissue, induce the inflammatory reaction of lung tissue and promote lung metastasis. Celecoxib can relieve lung inflammation and inhibit lung metastasis.