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机构地区:[1]湖北医药学院附属人民医院妇产科,湖北十堰442000
出 处:《南京医科大学学报(自然科学版)》2018年第1期44-48,共5页Journal of Nanjing Medical University(Natural Sciences)
摘 要:目的:研究CD146基因在卵巢癌细胞SKOV3中过表达对其接种裸鼠皮下生成肿瘤能力的影响。方法:以脂质体介导的CD146基因转染卵巢癌细胞SKOV3,G418筛选阳性克隆细胞,并经Western blot鉴定其过表达CD146蛋白。分别应用MTT与软琼脂克隆形成试验评价过表达CD146克隆细胞对细胞增殖与体外集落形成能力的影响,并将这些细胞接种裸鼠皮下组织,每周观察、记录肿瘤的生长状况至裸鼠处死,称量肿瘤重量并对肿瘤组织进行免疫组织化学染色观察。结果:SKOV3细胞经脂质体转染与G418筛选后,可获得过表达CD146的阳性克隆细胞。CD146过表达明显抑制了SKOV3细胞的体外增殖能力与集落形成能力,亦明显抑制了裸鼠皮下的肿瘤生成能力,其肿瘤重量与体积明显比对照组小(P<0.05)。免疫组织化学染色结果表明过表达CD146的SKOV3细胞接种裸鼠生成的肿瘤组织仍可高表达CD146蛋白分子。结论:CD146过表达明显抑制卵巢癌细胞SKOV3的体外增殖能力与裸鼠体内的致瘤性,可能是卵巢癌预后的一个潜在分子标记。Objective: To explore the effects of CD146 overexpression in ovarian cancer cell line SKOV3 on the tumorigenesis in nude mice. Methods: The lipofection-mediated transfection in SKOV3 cells was performed and transfectants were screened by G418 resistance to obtain positive clone cells that overexpressed CD146 protein confirmed by Western blot. MTT assay and colony formation assay on soft agar were used to evaluate the proliferation and colony formation ability of SKOV3 cells that overexpressed CD146, respectively.These positive cells were subcutaneously injected into the nude mice, and the tumorigenesis of SKOV3 was observed and recorded every week until the sacrifice of the nude mice. The tumor weight and volume were measured and immunohistochemical staining was used to detect CD146 expression in the tumor tissue of nude mice. Results: After lipofection and G418 screening, the positive clone cells overexpressed CD146 were selected. CD146 overexpression significantly inhibited cell proliferation and decreased colony formation ability in SKOV3 cells in vitro, and also inhibited the tumorigenesis of SKOV3 in the nude mice. The tumor weight and volume from the nude mice injected subcutaneously with overexpressed-CD146 SKOV3 cells were respectively lighter and smaller than those of the control(P〈0.05). Immunohistochemical staining showed that tumor tissues from nude mice injected with overexpressed-CD146 SKOV3 cells still could express high level of CD146 protein. Conclusion: CD146 overexpression can inhibit cell proliferation in vitro and tumorigenesis of SKOV3 in nude mice. CD146 may be a potential marker for ovarian cancer prognosis.
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