Résumé
Background and purpose:Insulin-like growth factor-1 (IGF-1) is a peptide that participates in many biological processes by stimulating the downstream signaling pathways through their interaction with IGF-1 re-ceptor (IGF-1R) and insulin receptor (IR). Bone morphogenetic proteins (BMPs) are a group of functional proteins which participate in the biological processes of proliferation and migration in many kinds of cancers and have become a hot area of cancer research. The study aimed to investigate the effects of silencingIGF-1R gene on the expression level ofBMP2 gene, and the cell proliferation and apoptosis of SMMC7721 cells.Methods:The RNAi plasmid targetingIGF-1R gene was constructed and transfected into SMMC7721 cells. Then the inhibition effect on the expression level of IGF-1R and BMP2 gene was detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot. The SMMC7721 growth curve and cell apoptosis were detected by MTT assay and flow cytometry after they were transfected with RNAi plasmid.Results:The RNAi plasmid targetingIGF-1R gene was constructed successfully. The inhibition efficiencies at mRNA expression levels were 68.9% and 80.7% (IGF-1R gene), 79.5% and 83.3% (BMP2 gene), respectively, after transfection with IGF-1R-siRNA-1 and IGF-1R-siRNA-2 plasmid (P<0.05). The inhibition efficiencies at protein levels were 46.1% and 62.1% (IGF-1R gene,P<0.05), 42.5% and 60.9% (BMP2 gene,P<0.05), respectively. The results of MTT growth curve showed that the proliferation rate in the transfected SMMC7721 cells was significantly slower than that in the control group (P<0.05). The proportion of apoptotic cells in transfected groups was significantly higher than that in the control group (P<0.05).Conclusion:SilencingIGF-1R gene can downregulate the expression ofBMP2 gene at different levels that results in inhibition of cell proliferation and promotion of apoptosis in SMMC7721 cells.
Résumé
Objective To discuss the effect of daidzein on hepatocellular carcinoma SMMC7721 cell proliferation CD133 expression on tumor stem cell.Methods Hepatocellular carcinoma SMMC7721 cells were cultured,digested and passaged,and divided into six groups with different drug:control group with no daidzein,100 μg/mL daidzein group,200μg/mL daidzein group,300μg/mL daidzein group,400μg/mL daidzein group ,500μg/mL daidzein.The inhibition ratio,hexokinase,alkaline phosphatase and CD133 levels in SMMC7721 cell were detected and compared at 24 h,48 h,72 h among those groups. Results The inhibition ratio was increased by Daidzein dose increasing,and decreased apparently by times extending,especially in 400μg/mL and 500μg/mL daidzeingroups.Compared with control group,the hexokinase and alkaline phosphatase activity and CD133 expression were decreased apparently in groups treated with daidzein(P<0.01).The more the dose,the higher the drop(P<0.01).Conclusion Daidzein can inhibit hepatocellular carcinoma SMMC7721 cell proliferation,and inhibit CD133 expression on tumor stem cell.