In vitro malignant transformation of human bronchial epithelial cells induced by benzo(a)pyrene.
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.
Department of Toxicology, Peking University Health Science Center, Beijing 100191, PR China.
Peng
P
Zhao
Juanling
J
Fu
Biyun
B
Yao
Yanchao
Y
Song
Lan
L
Mi
Zhenning
Z
Li
Lanqin
L
Shang
Weidong
W
Hao
Zongcan
Z
Zhou
D000818:Animals; D001564:Benzo(a)pyrene / Q000633:toxicity*; D001980:Bronchi / Q000166:cytology; D002273:Carcinogens / Q000633:toxicity*; D002460:Cell Line; D002465:Cell Movement; D002471:Cell Transformation, Neoplastic / Q000139:chemically induced* / Q000378:metabolism / Q000473:pathology; D004847:Epithelial Cells / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D006801:Humans; D008175:Lung Neoplasms / Q000556:secondary; D051379:Mice; D008819:Mice, Nude; D009363:Neoplasm Proteins / Q000378:metabolism; D009368:Neoplasm Transplantation; D009369:Neoplasms / Q000473:pathology; D018809:Proliferating Cell Nuclear Antigen / Q000378:metabolism; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism; D047368:Tumor Burden; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism
D002273:Carcinogens; D009363:Neoplasm Proteins; D018809:Proliferating Cell Nuclear Antigen; C114751:TPD52 protein, human; D016159:Tumor Suppressor Protein p53; D001564:Benzo(a)pyrene; D051057:Proto-Oncogene Proteins c-akt