The purpose of this study was to investigate the potential role of PHRF1 in lung tumorigenesis. proliferation, colony formation and growth of tumor xenograft and = 0.009) (Figure ?(Figure1A1A). Open in a separate window Physique 1 The relative level of PHRF1 mRNA expression in human lung tissue was discovered by qRT-PCR as well as the appearance of PHRF1 in lung tumor cell lines and malignant 16HEnd up being cells induced by BaP was discovered by traditional western blotThe lower degree of PHRF1 mRNA appearance was seen in individual lung tumor tissue than that in paracancerous tissue A. The reduced appearance of PHRF1 proteins was seen in lung cancer cell lines (H1299 and H1650) than that in normal human bronchial epithelial cell lines (16HBE and BEAS-2B) B. The decreased expression of PHRF1 protein was observed in malignant 16HBE cells induced by BaP than that in control cells C. The expression of PHRF1 in lung cancer cell lines and malignant 16HBE cells induced by BaP The significantly lower level of PHRF1 mRNA expression was observed in human lung cancer tissues than that in paracancerous tissues, thus, we detected the expression of PHRF1 in lung cancer cell lines (H1299 and H1650) and in normal human bronchial epithelial cell lines (16HBE and BEAS-2B) further. Western blot analysis showed that this markedly decreased expression of PHRF1 protein was observed in H1650 and H1299 cell lines compared with 16HBE and BEAS-2B cell lines (Physique ?(Figure1B).1B). Malignant 16HBE cell line induced by BaP was reported in our previous paper, which could form colonies in soft agar and grow tumor in nude mice [13]. Western blot analysis showed that the dramatically reduced expression of PHRF1 protein was observed in 16HBE-BaP cells as compared to that in 16HBE-control cells (Physique ?(Physique1C1C). The expression of PHRF1 in mice lung tissues treated by BaP Above, we observed the markedly reduced expression of PHRF1 in lung cancer from population study and experimental studies as compared to those in its corresponding controls. To research the design of PHRF1 appearance in test further, feminine Kunming mice had been treated with BaP to stimulate lung tumorigenesis. Traditional western blot evaluation indicated the fact that significantly decreased appearance of PHRF1 proteins was seen in mice lung tissue treated with BaP than that in charge group (Body ?(Figure2A).2A). The immunohistochemistry assay demonstrated the fact that staining of PHRF1 was weakened in BaP-treated group, but solid in charge group (Body ?(Figure2B2B). Open up in another window Body 2 The appearance of PHRF1 in BaP-treated mice lung tissue was assessed by traditional western blot assay and immunohistochemistryThe reduced appearance of PHRF1 proteins was seen in BaP-treated mice lung tissue weighed against control group A. B. demonstrated the fact that staining of PHRF1 was weakened in BaP-treated group, but solid in charge group. The consequences of overexpression of PHRF1 on H1299 cell proliferation To see the consequences of overexpression of PHRF1 in the cell proliferation, H1299 cells had been contaminated with Nrp2 PHRF1 lentiviruses. Body ?Body3A3A showed an H1299 cell range stably overexpressing PHRF1 was successfully established. The cellular number was MIK665 counted at different period points. As proven in Figure ?Body3B,3B, the growth of H1299-PHRF1 cells was slower than that of H1299-pvoid cells from 72h ( 0 significantly.05). Open up in another window Physique 3 The effects of overexpression of PHRF1 on H1299 cell proliferationA. Western blot analysis showed that a stable PHRF1-overexpressed H1299 cell collection was established. B. Overexpression of PHRF1 inhibited H1299 cell proliferation from 72h. * 0.05). Open in a separate window Physique 4 The effects of overexpression of PHRF1 on colony formation and tumor growth in mouse xenograft modelThe MIK665 dramatically decreased colony formation was observed in H1299-PHRF1 cells as compared to H1299-pvoid cells A. There was a significant reduction in the mean of tumor excess weight in H1299-PHRF1 cells as compared to H1299-pvoid cells B. * 0.05), which suggests that overexpression of PHRF1 significantly inhibited the tumor formation and tumor growth of H1299 cells 0.05) and the percentage of S phase cells was significant lower in H1299-PHRF1 cells (32.72.7%) than that in H1299-pvoid cells (45.84.7%) ( 0.05) (Figure ?(Figure5),5), which suggests that overexpression of PHRF1 could induce the growth inhibition of H1299 cells by arresting the cell cycle in G1 phase. In addition, we observed that this proliferation index was significantly decreased in H1299-PHRF1 cells (41.43.9%) MIK665 than that in H1299-pvoid cells (55.23.3%) ( 0.05). Open in a separate window Physique 5 The effects of overexpression of PHRF1 around the distribution of cell cycleFlow cytometry analysis showed that overexpression of PHRF1 increased the cell percentage in G1 phase and decreased the cell percentage in S.