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目的:建立人前列腺癌LNCap细胞高成瘤性亚克隆细胞株并探讨其生物学特性。方法:慢病毒载体PGK-luciferase-GFP转染LNCap细胞,稳转株与Matrigel混合注入SCID小鼠皮下,反复消化法原代培养皮下瘤获得亚克隆细胞株,CCK8及Transwell实验检测细胞增殖及迁移力;用亲代LNCap细胞及亚克隆细胞皮下注射SCID小鼠成瘤,观察两组瘤体生长及大小,并用生物活体发光、病理及免疫组织化学检测瘤体情况。结果:与亲代LNCap细胞相比,亚克隆细胞的生长速度增快,迁移力增强,皮下瘤成瘤率增高,荧光信号强度增加,皮下瘤组织切片HE染色镜下可见核分裂相,免疫组织化学AR染色阳性。结论:制备的人前列腺癌亚克隆细胞株其恶性生物行为增加。
OBJECTIVE: To establish a highly tumorigenic subclone cell line of human prostate cancer LNCap cells and investigate its biological characteristics. METHODS: LNCap cells were transfected with lentivirus vector PGK-luciferase-GFP. The metastatic cells were mixed with Matrigel and subcutaneously injected into SCID mice. Subclinical cells were subcultured by repeated digestion method to obtain subclone cell lines. CCK8 and Transwell assay were used to detect cell proliferation and migration The SCID mice were injected subcutaneously with LNCap cells and subclone cells. The tumor growth and size of the two groups were observed. The growth and size of the tumors were observed and the tumors were detected by bioluminescence, histopathology and immunohistochemistry. Results: Compared with the parental LNCap cells, the growth rate of subclone cells was increased, the migration ability was enhanced, the tumorigenic rate of hypoxia tumor increased, and the fluorescence signal intensity was increased. The mitotic phase was observed under the HE staining of subcutaneous tumor tissue sections. Immunohistochemistry AR Positive staining. Conclusion: The human prostate cancer subclone cell line has increased malignant biological behavior.