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Objective To study the effect and mechanisms of TW-37 on cell proliferation,apoptosis,invasion and angiogenesis in pancreatic cancer cells in vitro and further explore the potential mechanism.Methods BxPC3 and HPAC cells were pretreated with TW-37 using untransfected or transfected with NF-κB p65 cDNA(p65 cDNA)or NF-κB p65 siRNA(siRNA-p65)cells as controls.Cell viability was determined by MrTT assay.Cell apoptosis was assessed by enzyme-linked immunosorbent assay (ELISA).Cell invasion and angiogenesis was detected by Transwell and endothelial tube formation assay of HUVECs.ELISA assay was used to measure the activity of NF-κB,and its target proteins of MMP-9 and VEGF were detected by western blot.Results TW-37 suppressed cell growth and induced apoptosis (A405:1.29 ± 0.21 vs 0.09 ± 0.01,1.07 s0.18 vs 0.08 ± 0.01),inhibited NF-κB activity and protein expression of NF-κB p65,VEGF and MMP-9(all P ＜0.05)in a dose-and time-dependent manner.The number of cells that invaded across the matrigel in the transwell chamber was (46.7 ±5.24) and (10.3 ± 1.26)/×200 in BxPC3 control and 0.75 μmol/L TW-37 group (P=0.001).The number of tube formation was (39.4 ±4.36) and (7.84 ± 1.25)/×200,(P =0.001).NF-κB activity was increased by p65 cDNA transfection,and decreased by TW-37 treatment in both of the two cell lines (P ＜0.05).However,NF-κB activity was decreased by p65 siRNA transfection,and greatly decreased by TW-37 treatment in both two cell lines (P ＜0.05 or P ＜0.01).Transfection of p65 cDNA did not significantly affect cell apoptosis.Transfection of p65 siRNA increased cell apoptosis,and greatly increased by TW-37 treatment in both two cell lines (all P ＜ 0.01).Conclusions TW-37 could inhibit the proliferation,invasion and angiogenesis in pancreatic cancer cells by regulating NF-κB signal pathway.
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<p><b>OBJECTIVE</b>To construct a lentiviral expression vector for human VHL and its shRNA vector, and study the effect of VHL on proliferation and apoptosis of renal cell carcinoma cell lines.</p><p><b>METHODS</b>Lentiviral vectors pZsGreen1-VHL and pLL3.7-shVHL were constructed and transfected into 293T cells with 3 packaging plasmids by Lipofectamine(TM) 2000 reagent. The supernatant was collected to infect A498 and Caki-1 cells, respectively. VHL mRNA and protein levels were detected by RT-PCR and Western blotting, respectively. The effect of VHL on the proliferation, cell cycle and cell apoptosis were analyzed by MTS and flow cytometry.</p><p><b>RESULTS</b>The recombinant lentiviral vectors were successfully constructed. The proliferation of A498 cells with reconstituted wild-type VHL was significantly inhibited, while the proliferation of Caki-1 cells with VHL knockdown was significantly enhanced as compared with the control cells (P<0.05). VHL induced G0/G1-S cell cycle arrest. The apoptosis rate of A498 cells with reconstituted wild-type VHL was significantly increased while that of Caki-1 cells with VHL knockdown was significantly lowered compared with the control cells (P<0.05).</p><p><b>CONCLUSION</b>VHL can inhibit the proliferation and induce apoptosis of renal cell carcinoma cells.</p>
Subject(s)
Humans , Apoptosis , Carcinoma, Renal Cell , Pathology , Cell Cycle , Cell Line, Tumor , Cell Proliferation , Genetic Vectors , Lentivirus , Plasmids , RNA, Messenger , RNA, Small Interfering , Transfection , Von Hippel-Lindau Tumor Suppressor Protein , GeneticsABSTRACT
0.05),whereas there was a significant difference in cardiac function in patients received atorvastatin(P
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0.05). Conclusions The antihypertensive effects of NAHC treatment program could keep the patients' blood pressure in the ideal range.