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目的:阐明小鼠去细胞拟胚体对小鼠Lewis肺癌细胞在体内生长的影响。方法:先制备来源于小鼠胚胎干细胞的拟胚体,然后用SDS去细胞处理。实验分成3组:小鼠Lewis肺癌细胞与去细胞拟胚体培养组,癌细胞与Matrigel培养组和单纯癌细胞组(每组n=12)。培养3天后注射入裸鼠体内,观察肿瘤生长情况。28天取出瘤体,Ki67和CD31免疫组化染色(n=12)检测细胞增殖和肿瘤微血管密度(MVD),Western blot检测组织Paxillin,E-cadherin和β-actin水平(n=6)。结果:去细胞拟胚体组肿瘤生长明显较单纯细胞组和Matrigel组慢。去细胞拟胚体组Ki67指数((17.1±2.6)%)明显小于单纯细胞组((34.5±4.7)%)和Matrigel组((48.4±8.6)%)(P<0.05);去细胞拟胚体组的MVD(18.7±3.6个/mm2)明显小于单纯细胞组(32.1±6.4个/mm2)和Matrigel组(42.6±7.1个/mm2)(P<0.05)。Western blot结果提示去细胞拟胚体组的Paxillin表达小于单纯细胞组和Matrigel组(P<0.05),而E-cadherin表达大于单纯细胞组和Matrigel组(P<0.05)。结论:小鼠去细胞拟胚体对小鼠Lewis肺癌细胞在体内有明显的促分化作用。
Objective: To clarify the effect of mouse embryoid body on mouse Lewis lung carcinoma cell growth in vivo. Methods: The embryoid bodies derived from mouse embryonic stem cells were prepared and then treated with SDS. The experiment was divided into three groups: mouse Lewis lung cancer cells and embryoid dendritic cell culture group, cancer cells and Matrigel culture group and simple cancer cell group (n = 12). After 3 days of inoculation into nude mice, the growth of the tumor was observed. The tumors were removed on day 28, and the cell proliferation and tumor microvessel density (MVD) were detected by immunohistochemical staining of Ki67 and CD31 (n = 12). The levels of Paxillin, E-cadherin and β-actin were detected by Western blot (n = 6). Results: The growth of tumor in acellular embryoid body was significantly slower than that in pure cell group and Matrigel group. The Ki67 index of decellularized embryoid body was (17.1 ± 2.6)%, which was significantly lower than that of pure cell group (34.5 ± 4.7%) and Matrigel group (48.4 ± 8.6%) (P <0.05) The MVD of the body group (18.7 ± 3.6 cells / mm2) was significantly less than that of the pure cell group (32.1 ± 6.4 cells / mm2) and the Matrigel group (42.6 ± 7.1 cells / mm2) (P <0.05). The results of Western blot showed that Paxillin expression in decellularized embryoid body was lower than that in pure cell group and Matrigel group (P <0.05), while E-cadherin expression was greater than that in pure cell group and Matrigel group (P <0.05). CONCLUSION: Mouse acellular embryoid bodies can promote the differentiation of Lewis lung carcinoma cells in vivo.