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Effect Of SiRNA Interference Targeting BMPR-Ⅱ Gene On Cell Proliferation And Invasion Ability Of Human Hepatoma HepG2Cells

Posted on:2014-03-02Degree:MasterType:Thesis
Country:ChinaCandidate:S CaiFull Text:PDF
GTID:2254330425458331Subject:Oncology
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Background and purpose:Primary hepatocellular carcinoma is one of the most common malignant tumorsin our country. Recent studies showed that bone morphogenetic proteins (BMPs)signaling pathways play a key role in the development of tumors. This study aimed toinvestigate the effect of small interfering RNA (siRNA) on the inhibiting of bonemorphogenetic protein receptor (BMPR-Ⅱ), and to observe its effect on theproliferation and invasion of human hepatoma HepG2cells after BMPR-Ⅱsilencing.Methods:Three pairs of specific siRNA targeting BMPR-Ⅱwere designed and synthesized,and then transiently transfected into HepG2cells via cathodolyte liposometransfection method. In vitro cultured HepG2cells were divided into a normal controlgroup, a blank control group, a negative control group and three transfected withsiRNA-BMPR-Ⅱ-a, siRNA-BMPR-Ⅱ-b, siRNA-BMPR-Ⅱ-c groups. RT-PCR andWestern blotting were used to detect BMPR-Ⅱ expression at mRNA and proteinlevels, and the proliferation and invasion ability of HepG2cells after transfectionwere assessed by MTT assay and Transwell invasion assay respectively.Results:48h after the three pairs of specific siRNA-BMPR-Ⅱs were transfected intoHepG2cells, RT-PCR and Western blotting results showed that the expression ofBMPR-Ⅱ mRNA and protein were significantly inhibited compared with those inthe other three groups, with siRNA-BMPR-Ⅱ-a having the highest inhibitory rate(0.70±0.06,0.45±0.10, P<0.05). MTT assay and Transwell invasion assaydemonstrated the growth and the number of HepG2cells in siRNA-BMPR-Ⅱ-agroup was significantly lower (48.27%±0.76%,25.20±1.60, P<0.05) than in normalcontrol group (82.64%±0.67%、60.40±1.39) and negative control group (81.21%±0.80%、59.50±1.85)48h after transfection. Conclusion:siRNA interference-mediated BMPR-Ⅱ gene silencing can inhibit theproliferation and invasion ability of human HepG2cells, and BMPR-Ⅱ may be usedas a new promising target of antineoplastic therapy in hepatocellular carcinoma.
Keywords/Search Tags:Hepatocellular carcinoma, RNA interference, Bone morphogeneticprotein receptorⅡ, Proliferation, Invasion
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