Study On Mechanism Of Sinomenine In Human Gastric Cancer Cells SGC-7901

Object: To investigate the role of SIN on proliferation, EMT-related m RNA, and protein expression of human gastric cancer SGC-7901 cells in vitro.Methods:1 Cell viability: Treated with different concentrations of SIN, the human gastric cancer SGC-7901 cells viability assay was performed with WST-1 after different treated time.2 MMPs expression: Treated with different concentrations of SIN, MMP-2 and MMP-9 content in the cell culture medium were detected with ELISA kit.3 QPCR detection: Treated with different concentrations of SIN, the influence of SIN to EMT-associated m RNA expression of human gastric carcinoma SGC-7901 cells was detected with RT-PCR.4 Westen blotting analysis: The change of EMT-associated protein levels of SGC-7901 cells after exposed with different concentrations SIN was detected by Western Blotting.5 NFκB p65 Westen blotting analysis The impact of SIN to nuclear NFκB p65 protein levels of SGC-7901 by Western blotting.Results:1 Treated with the different concentration of SIN, the human gastric cancer SGC-7901 cells showed that: the cell survival in 1 mmol/L, 2 mmol/L, 2.5 mmol/L and 3 mmol/L treatment groups was significantly lower than the control group(P<0.05) after 24 h treatment, while the cell survival in 0.5 mmol/L, 1 mmol/L, 2 mmol/L, 2.5 mmol/L and 3 mmol/L treatment groups was significantly lower than the control group(P<0.05) after 48 h treatment, with time and dose dependent manner to SIN.2 The MMPs levels of human gastric cancer SGC-7901 cell culture medium were decreased after SIN treatment. The MMP-2 level in each SIN group medium was significantly lower than the control group(P<0.05), and MMP-9 content of 1 mmol/L and 2 mmol/L groups were reduced significantly(P<0.05).3 RT-PCR and Western Blotting results showed that: in contrast to the control group, the MMP-2 m RNA and protein expression, MMP-9 m RNA expression of each treatment group were significantly decreased(P<0.05) with increasing SIN concentration. In the meanwhile, the MMP-9 protein in 0.5 mmol/L, 1 mmol/L and 2 mmol/L treatment group was significantly lower(P<0.05). Snail m RNA and protein levels in 0.5 mmol/L, 1 mmol/L and 2 mmol/L treatment groups were significantly decreased(P<0.05) compared to the control group, and E-cadherin gene and protein levels were significantly increased(P<0.05) in 0.5 mmol/L, 1 mmol/L and 2 mmol/L treatment groups.4 The Western Blotting results showed that with the SIN concentration increased, NFκB p65 protein in SGC-7901 cell nuclei showing a decreasing trend. And 0.5 mmol/L, 1 mmol/L and 2 mmol/L treatment groups were significantly lower compared to the control group(P< 0.05).Conclusion: SIN can inhibit the proliferation of gastric cancer cell line SGC-7901 with a time and concentration dependent manner. SIN can regulate the EMT –related protein expression and the nuclear translocation of NFκB p65 in gastric cancer cell line SGC-7901, suggesting that the nuclear translocation of NFκB p65 may be involved in the SIN regulation of EMT in SGC-7901.