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Archive > Volume 22 Issue 6 > 2015,22(6):696-702. DOI:10.3872/j.issn.1007-385X.2015.06.004 Prev Next
Monographic Report
The inhibitory effect of siRNA silencing NF-κB gene family members on the expressions of NKG2DLs on human hepatocellular carcinoma HepG2 cells
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Abstract:
Objective: To investigate the interaction between the natural killer group 2 member D ligands (NKG2DLs) expressions and NF-κB signaling pathway in HepG2 cells treated with sunitinib.Methods: HepG2 cells were cultivated by routine method. The mRNAs of NF-κB gene family members in HepG2 cells were quantitated by RT-qPCR. The siRNAs specific for NF-κB1, NF-κB2 and RelB were designed using a computer soft and synthesized. They were transfected into HepG2 cells using liposome. The transfection and interference efficacy were evaluated by fluorescence microscopy, and real-time quantification PCR respectively. The expressions of NF-κB1, NF-κB2 and RelB were determined by immunoblotting, and the expressions of NKG2DLs were assessed by Flow cytometry.Results: The mRNAs of NF-κB1, and especially NF-κB2 and RelB were significantly increased in HepG2 cells treated with sunitinib. In siRNA transfection experiments, the red fluorescence was present in about 60% of HepG2 cells under fluorescence microscope, and the interference efficiency was 95%. Immunoblotting revealed that NF-κB1, NF-κB2 and RelB were decreased significantly in HepG2 cells after the transfection. Flow cytometry analysis found that the expressions of NKG2DLs in the siRNA transfected group was significantly lower than that in drug treatment group (P<005). Conclusion: Sunitinib induces the expressions of NKG2DLs on hepatocellular carcinoma HepG2 cells by the activating the alternative NF-κB pathway.
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Project Supported:
Project supported by the National Natural Science Foundation for Young Scientists of China (No. 81302372，81300431), and the Excellent Middle and Yong Aged Experts Project of Zhujiang Hospital Southern Medical University（No.201207008 )
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