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Objective:To explore the inhihitive and apoptosis inductive effect of IL-24 genes on CD133~+laryngeal cancer cells in Hep-2 line.Methods:Human peripheral blood monocytes were isolated.The total RNA was extracted by using Trizol method and reverse transcripted into cDNA using RT-PCR method.Primers P1 and P2 was designed for the amplification of human IL-24 genes.After confirmation of agarose gel electrophoresis tests,TA was cloned into pMD19-T simple vector.Nhe Ⅰ and Xho Ⅰ double digesting human IL-24 and pIRES2-ZsGreen1 and eukaryotic expression vector were used to establish the pIRES2-ZsGreen1-hIL-24 vector,and detected by enzyme digestion and gene sequencing methods.Flow cytometry(FCM) was used to isolate CD133~+ cells from Hep-2 cells.CD133~+ cells were transfected with pIRES2-ZsGreen1-hIL-24 through liposome 2000.After detection,MTT and FCM were used to observe the effect of IL-24 gene on CD133~+ laryngeal cancer Hep-2 cells.Results:Lipotin mediated transfection of recombinant pIRES2-ZsGreen1-hIL-24 plasmid into CD133~+ Hep-2 could expressed IL-24 gene in cells stably.MTT results showed that IL-24 transfected group was significantly suppressed compared to empty vector group and control group(P<0.05);FCM results showed that the apoptosis rate of experimental group increased significantly compared to empty vector group and control group(P<0.05).Conclusions:IL-24 gene expressions can inhibit proliferation of CD133~+laryngeal cells in Hep-2 line and promote their apoptosis.
Objective: To explore the inhihitive and apoptosis inductive effect of IL-24 genes on CD133 ~ + laryngeal cancer cells in Hep-2 line. Methods: Human peripheral blood monocytes were isolated. The total RNA was extracted by using Trizol method and reverse transcripted into cDNA using RT-PCR method. Primers P1 and P2 were designed for the amplification of human IL-24 genes.After confirmation of agarose gel electrophoresis tests, TA was cloned into pMD19-T simple vector. Nhe I and Xho I double digesting human IL -24 and pIRES2-ZsGreen1 and eukaryotic expression vector were used to establish the pIRES2-ZsGreen1-hIL-24 vector, and detected by enzyme digestion and gene sequencing methods. Flow cytometry (FCM) was used to isolate CD133 ~ + cells from Hep- 2 cells.CD133 ~ + cells were transfected with pIRES2-ZsGreen1-hIL-24 through liposome 2000. After detection, MTT and FCM were used to observe the effect of IL-24 gene on CD133 ~ + laryngeal cancer Hep-2 cells. Results : Lipotin mediated transfection of recombinant The results of IL-24 transfected group were significantly suppressed compared to empty vector group and control group (P <0.05). The expression of pIRES2-ZsGreen1-hIL-24 plasmid into CD133 ~ + Hep- ; FCM results showed that the apoptosis rate of experimental group increased significantly compared to empty vector group and control group (P <0.05) .Conclusions: IL-24 gene expressions can inhibit proliferation of CD133 ~ + laryngeal cells in Hep-2 line and promote their apoptosis.