ABSTRACT
Objective:To investigate the effects of Mor-platin, a novel mitochondrial platinum complex, on proliferation and migration of human hepatoma carcinoma HepG2 cells. Methods:Cell counting kit-8 (CCK-8) assay was used to analyze cell proliferation of Mor-platin and classic anticancer drugs, particularly cisplatin, in HepG2 cells. A laser confocal microscope was used to observe whether Mor-platin can target mitochondria. The morphological changes in cellular mitochondria after treatment with Mor-platin were ob-served on a transmission electron microscope. Cell apoptosis was measured by flow cytometry, and cell invasion was evaluated by three-dimensional tumor spheroid model. Results:Mor-platin can inhibit cell proliferation and is dose dependent. The half inhibitory concentration (IC50) of Mor-platin is lower than that of cisplatin. Laser confocal images showed that Mor-platin can target cell mito-chondria and enrich cell mitochondria. Transmission electron microscopy images showed that cell mitochondrial morphology changed after Mor-platin treatment. Furthermore, cell mitochondrial membrane is incomplete and mitochondrial cristae are reduced. Cell apoptosis caused by Mor-platin is dose dependent. The three-dimensional tumor spheroid model showed that the cell areas of the group subjected to Mor-platin treatment are smaller than those of the control group. Conclusion:Mor-platin can target cell mitochon-dria, change the cell mitochondrial morphology, inhibit cell proliferation, and thus promote cell apoptosis. It also showed better anti-cancer effects than cisplatin. Furthermore, Mor-platin can inhibit three-dimensional tumor spheroid invasion. These results suggest that Mor-platin is a potential antitumor drug.
ABSTRACT
Objective To investigate the abnormal expression of Golgi protein 73(GP73) in CD4+ T lymphocytes of the pa-tients with hepatocellular carcinoma (HCC) and its influence of Th1/Th2/Th17 subtype differentiation .Methods Fifty cases of HCC hospitalized in this hospital from May 2015 to February 2016 and 50 healthy volunteers as controls were selected .Peripheral blood was collected and CD4+ T lymphocytes were isolated ;then the expression levels of GP73 and nuclear factor kappa-light-chain-enhancer (NF-κB) in CD4+ T lymphocytes were determined by using RT-qPCR and Western blotting methods ;furthermore ,the se-cretion levels of IL-4 ,IL-17 and IFN-γin the supernatants were examined by using ELISA method .Results GP73 mRNA expres-sion in peripheral blood CD4+ T lymphocytes in the HCC patients were significantly up-regulated compared with the healthy volun-teers ,the difference was statistical difference (P<0 .05) .The expression level of in GP73 overexpression group was significantly in-creased(P<0 .05) ,while which in the GP73 interference group was significantly decreased (P<0 .05) .Over-expression of GP73 in-duced significant increase of IL-4 and IL-17 levels and significant decrease of IFN-γ(P<0 .05);silencing GP73 induced marked de-crease in the expression of IL-4 and IL-17 in CD4+ cells and obvious increase of IFN-γ(P<0 .05) .Conclusion GP73 is over-ex-pressed in peripheral blood CD4+ T cells of HCC patients ,moreover GP73 is very likely to participate in the inflammatory reaction by activating NF-κB to cause the unbalance of Th1/Th2/T17 and promote the occurrence and development of HCC .
ABSTRACT
Objective To investigate the effect of deltaNp63(ΔNp63) silencing on the proliferation of pancreatic cancer PANC1 cells.Methods ΔNp63 mRNA level in 23 pairs of pancreatic cancer and adjacent tissue specimen was detected by real-time PCR, andΔNp63 protein in human normal pancreatic ductal cell line HPDE6-C7 and pancreatic cancer cell line PANC1, CFPAC1 and BXPC3 was detected by Western blot. PANC1 cells were transfectedΔNp63 specific siRNA (ΔNp63-siRNA ) and scramble siRNA ( Con-siRNA ) using liposome, and untransfected cells served as control.ΔNp63 mRNA and protein was detected by real-time PCR and Western blot to validate the silencing ofΔNp63 expression.MTT assay and BrdU method were used to detect the proliferation and DNA synthesis of transfected PANC1 cells.Results TheΔNp63 mRNA expression in pancreatic cancer tissues and matched adjacent normal tissues was 0.99 ± 0.07 and 0.70 ±0.07, respectively.ΔNp63 mRNA expression in pancreatic cancer tissue was significantly up-regulated compared with that in the normal tissue (P=0.0034).TheΔNp63 protein expression in HPDE6-C7, PANC1, CFPAC-1 and BxPC3 cells was 0.97 ±0.09,3.06 ±0.16,2.57 ±0.11 and 2.45 ±0.08, respectively.TheΔNp63 protein level in pancreatic cancer cells were higher than that in HPDE6-C7 cells (P<0.001).ΔNp63 mRNA level in control, Con-siRNA and ΔNp63-siRNA group was 0.97 ±0.07,0.97 ±0.07 and 0.28 ±0.03, respectively, andΔNp63 protein expression level was 0.97 ±0.06,1.00 ±0.10 and 0.26 ±0.03.The expression ofΔNp63 mRNA and protein inΔNp63-siRNA group were significantly down-regulated comparing with those in Con-siRNA group (P<0.01).Significant inhibition on cell proliferation was observed in ΔNp63-siRNA group, which was statistically different from that in control and Con-siRNA group.The A490 value (DNA synthesis) of control, Con-siRNA andΔNp63-siRNA group was 0.55 ±0.04, 0.56 ±0.01 and 0.55 ±0.00 at 24 h after transfection, and 0.84 ±0.05,0.87 ±0.07 and 0.71 ±0.05 at 48 h after transfection.The DNA synthesis inΔNp63-siRNA group was significantly down-regulated compared with that in control and Con-siRNA group (P<0.05).Conclusions Knockdown ofΔNp63 could greatly inhibit the proliferation and DNA synthesis of pancreatic cancer PANC1 cells.