DAPT suppresses the proliferation of human glioma cell line SHG-44.

OBJECTIVE: To explore the suppressing effect of γ-secretase inhibitor DAPT on proliferation of human glioma cell line SHG-44 in vitro and its mechanism. METHODS: The SHG-44 cell was treated by DAPT with different concentration. The proliferation of cells was detected by MTT assay; cell cycle and TSC of CD133(+) were determined by flow cytometry analysis technique; the key factor in Notch signaling pathway (Notch-1, Delta-1, Hes-1) was measured by reverse transcriptase-polymerase chain reaction and western blotting. RESULTS: DAPT inhibited the growth and proliferation of SHG-44 cells significantly(P<0.05). And the inhibiting effect on SHG-44 cells produced by DAPT showed a dose-dependent manner. DAPT increased the rate of cells in G0/G1 phase of SHG-44 cells, while it decreased the rate of cells in S phase. TSC of CD133(+) was significantly reduced after DAPT treated SHG-44 cells. The expression of protein and mRNA of Notch-1, Delta-1 and Hes-1 were gradually downregulated with the increase of DAPT doses. CONCLUSIONS: DAPT can downregulate these key factor in Notch signaling pathway, reduce the TSC of CD133+ and inhibit the proliferation of SHG-44 cells.

DAPT suppresses the proliferation of human glioma cell line SHG-44

OBJECTIVE@#To explore the suppressing effect of γ-secretase inhibitor DAPT on proliferation of human glioma cell line SHG-44 in vitro and its mechanism.@*METHODS@#The SHG-44 cell was treated by DAPT with different concentration. The proliferation of cells was detected by MTT assay; cell cycle and TSC of CD133(+) were determined by flow cytometry analysis technique; the key factor in Notch signaling pathway (Notch-1, Delta-1, Hes-1) was measured by reverse transcriptase-polymerase chain reaction and western blotting.@*RESULTS@#DAPT inhibited the growth and proliferation of SHG-44 cells significantly(P<0.05). And the inhibiting effect on SHG-44 cells produced by DAPT showed a dose-dependent manner. DAPT increased the rate of cells in G0/G1 phase of SHG-44 cells, while it decreased the rate of cells in S phase. TSC of CD133(+) was significantly reduced after DAPT treated SHG-44 cells. The expression of protein and mRNA of Notch-1, Delta-1 and Hes-1 were gradually downregulated with the increase of DAPT doses.@*CONCLUSIONS@#DAPT can downregulate these key factor in Notch signaling pathway, reduce the TSC of CD133+ and inhibit the proliferation of SHG-44 cells.

Radio-sensitization of SHG44 glioma cells by Aidi injection in vitro.

In this study, we aimed to explore the radio-sensitization of the SHG44 glioma cell line by Aidi injection and the possible mechanisms involved. The growth curve, cloning efficiency and divisional index of the SHG44 cell line were observed. The inhibition ratio was determined by MTT assay, the change in the cell cycle was analyzed by flow cytometry and the expression of cyclin B1 and Wee1 was detected by western blot analysis. The reproductive activity of the group treated with irradiation (IR) and Aidi injection was suppressed significantly, and the cloning efficiency and divisional index also declined. Aidi injection (15 µg/ml) induced G2/M phase arrest efficiently in the cell line after 48 h. The expression of cyclin B1 decreased in the group treated with IR and Aidi injection compared with either of those with IR or Aidi injection alone. The expression of Wee1 increased in the group treated with IR and Aidi injection compared with that in the groups treated with either IR or Aidi injection alone. In conclusion, Aidi injection is effective in radio-sensitization. The possible mechanisms involved may be associated with G2/M phase cell arrest, the downregulation of cyclin B1 and upregulation of Wee1 expression.

Establishment of SHG-44 cell line stably and highly expressing MCHR2 / 第二军医大学学报

Objective: To construct a eukaryotic expressing vector harboring human melanin-concentrating hormone receptor 2 (MCHR2) and to establish a SHG-44 cell line stably and highly expressing MCHR2. Methods: The full-length MCHR2 cDNA fragment was amplified from the human fetal brain cDNA library by PCR and was cloned into pcDNA3.1(+) to construct eukaryotic vector pcDNA3.1(+)/MCHR2; the latter was then transduced into SHG-44 cells by Lipofectamine™. After screening culture by G418, SHG-44 cells stably expressing MCHR2 were established. The transcription and expression of MCHR2 was identified by RT-PCR, Western blotting and immunofluorescence. Results: The full-length MCHR2 cDNA fragment was amplified and the eukaryotic expression vector pcDNA3.1(+)/MCHR2 was successfully constructed. The expression of MCHR2 was found positive by RT-PCR, Western blotting and immunofluorescence, indicating that the SHG-44 cell line stably and highly expressing MCHR2 was successfully established. Conclusion: The successful establishment of MCHR2-SHG-44 cell line provides a solid foundation for further study on MCHR2 function.