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- W2378605051 abstract "Background and purpose: The human oncogene B-cell-specifi c moloney murine leukemia virus integration site 1(Bmi-1) is an important member of the polycomb group family, and it regulates cell proliferation and senescence via INK4a/ARF locus. This study investigated the effects of Bmi-1-siRNA on the proliferation of lung adenocarcinoma cell line SPC-A1 cells with INK4a/ARF locus and clarify the mechanism of Bmi-1-mediated effect on proliferation of lung adenocarcinoma cells. Methods: In this study, we chose the most effi cient siRNA chain the pGeneshl-2-Bmi-1 sense-1 and inserted into a pSUPER-retro-neo retroviral vector. The packaged si-Bmi-1 pSUPERretro-neo retroviral vector was stably transfected into lung adenocarcinoma SPC-A1 cell line. The stably transfected cells were cultured and passed. After transfection, the levels of Bmi-1 mRNA and protein expression of SPC-A1 cells wereanalyzed by RT-PCR and Western blot respectively. Trypan blue, MTT and plate colony forming assay were performed to observe the proliferation capibility of SPC-A1 cells and evaluate the cloning forming ability in vitro. The potency of tumorigenesis was observed in nude mouse through hypodermic inoculation of SPC-A1 cells. Cell cycle distribution was analyzed by fl ow cytometry(FCM) in SPC-A1 cells. The expression levels of proliferation proteins including p16INK4 a, p53, Cyclin D1, PTEN, Akt and Ser473p-Akt were analyzed by Western blot. Results: The mRNA and protein expression levels of Bmi-1 were signifi cantly reduced in SPC-A1-Bmi-1-siRNA cells transfected with pSUPER-retroneo retroviral vector. Knockdown of Bmi-1 could inhibit the growth, colony formation in vitro and tumorigenesis in vivo of SPC-A1 cells(P0.01). The transfected SPC-A1 cells were arrested in G1 phase [(64.6±1.2)%, P0.05]. Compared with two control groups, p16INK4 a, p53 and Akt were not affected(P0.05), while Cyclin D1 and Ser473 pAkt were downregulated(P0.01) and PTEN was up-regulated(P0.01) in the SPC-A1-Bmi-1-siRNA cells. SPC-A1-Bmi-1-siRNA cells were treated with various concentrations of PTEN inhibitor to determine expression levels of PTEN, Bmi-1 and Ser473p-Akt protein. Ablation of PTEN rescued Bmi-1 and Ser473p-Akt expression in SPC-A1-Bmi-1-siRNA cells. Conclusion: Knockdown of Bmi-1 gene can arrest the proliferation of SPC-A1 cells through G0/G1 phase arrest by inhibiting Cyclin D1 expression indirectly, which may be not associated with p16INK4 a signaling pathway." @default.
- W2378605051 created "2016-06-24" @default.
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- W2378605051 date "2014-01-01" @default.
- W2378605051 modified "2023-09-23" @default.
- W2378605051 title "Effects of Bmi-1-siRNA on proliferation of lung adenocarcinoma SPC-A1 cells and its mechanism" @default.
- W2378605051 hasPublicationYear "2014" @default.
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