Department of Toxicology, Peking University Health Science Center, Beijing 100191, PR China.
Toxicol In Vitro. 2012 Mar;26(2):362-8. doi: 10.1016/j.tiv.2011.12.013. Epub 2011 Dec 29.
In this study, the human bronchial epithelial cells (16HBE) were treated five times with 10μM benzo(a)pyrene (BaP), followed by 20 passages culture, and the in vitro BaP-induced malignant transformation of 16HBE cells was established. Five colonies in soft agarose were then amplified and donated as T-16HBE-C1∼5 cells, respectively. T-16HBE-C1∼5 cells can form tumors subcutaneously in nude mice. Histopathological changes in the tumors indicated nests growth, high nuclear-cytoplasmic ratios, coarse and clumped chromatin, numerous and distinctly atypical mitoses, cell necrosis and surrounding normal adipose, muscle and connective tissue immersed. In addition, lung metastasis was observed in nude mice in T-16HBE-C1, 3 and 4 groups. In vitro cell migration assay results indicated that T-16HBE-C2∼5 cells showed much lower migration capabilities than 16HBE cells. Western blotting analysis showed that the expressions of p53 and p-Akt (Ser473) in T-16HBE-C1∼5 cells were significant higher than those in 16HBE cells. Our results demonstrated that BaP could induce the malignant transformation of 16HBE cells, and p53 and p-Akt (Ser473) might play crucial roles in BaP-induced carcinogenesis. The five monoclonal cell lines (T-16HBE-C1∼5) with different migration capabilities could be used as research models for further understanding the mechanisms of BaP-induced carcinogenesis and cell migration.
在这项研究中，用人支气管上皮细胞（16HBE）经 10μM 苯并(a)芘（BaP）处理 5 次，然后进行 20 代传代培养，建立 16HBE 细胞体外 BaP 诱导恶性转化。然后从软琼脂中扩增 5 个菌落，并分别命名为 T-16HBE-C1∼5 细胞。T-16HBE-C1∼5 细胞可以在裸鼠中皮下形成肿瘤。肿瘤的组织病理学变化表明巢状生长、高核浆比、粗糙和块状染色质、大量明显非典型有丝分裂、细胞坏死和周围正常脂肪、肌肉和结缔组织浸润。此外，在 T-16HBE-C1、3 和 4 组裸鼠中观察到肺转移。体外细胞迁移实验结果表明，T-16HBE-C2∼5 细胞的迁移能力明显低于 16HBE 细胞。Western blot 分析表明，T-16HBE-C1∼5 细胞中 p53 和 p-Akt（Ser473）的表达明显高于 16HBE 细胞。我们的结果表明，BaP 可诱导 16HBE 细胞恶性转化，p53 和 p-Akt（Ser473）可能在 BaP 诱导的致癌作用中发挥关键作用。具有不同迁移能力的五个单克隆细胞系（T-16HBE-C1∼5）可作为进一步研究 BaP 诱导致癌和细胞迁移机制的研究模型。
