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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.
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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.
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Objective To investigate the effects of internal and external biliary drainage on liver regeneration of the obstructive jaundice rats after partial hepatectomy.Methods The rat models of obstructive jaundice with 70％ liver resection were successfully constructed.All the 120 rats were randomly divided into the control group:rats received middle and left hepatic lobectomy; internal drainage group:a drainage tube was placed between the dilated bile duct and the duodenum; external drainage group:a drainage tube was placed in the dilated bile duct.There were 40 rats in each group.Rats in the internal and external drainage groups received middle and left hepatic lobectomy at postoperative day 7.The blood and hepatic tissues were collected at postoperative day 0,1,2,4,12,24,48,72 hours after operation,and the rate of liver regeneration and mitotic index were determined.The expression of proliferating cell nuclear antigen (PCNA) and signal transducer and activator of transcription 3 (STAT3) in the hepatic tissues were detected by immunohistochemistry,and the levels of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) were detected by ELISA,and the mRNA expressions of TNF-α and IL-6 were detected by RT-PCR.All data were analyzed using the one way analysis of variance or SNK test.Results Within 72 hours after partial hepatectomy,the rate of liver regeneration of the internal drainage group was 94.86％± 12.72％,which was significant higher than 62.39％±8.01％ of the external drainage group and 45.77％ ± 5.41％ of the control group (F =33.62,P ＜ 0.05).The mitotoic index and PCNA levels of the 3 groups had obvious increase at postoperative hour 12,and the mitotoic index and PCNA levels of the external drainage group reached peak at postoperative hour 24,which were 24.47％ ± 4.01％ and 88.1％ ± 9.2％,respectively,the mitotoic index and PCNA levels of the control group and the external drainage group reached peak at postoperative hour 48,which were 15.80％ ± 1.08％,58.3％ ± 5.8％ and 18.40％ ± 1.12％,70.2％ ± 6.9％,respectively.The mitotoic index and PCNA levels of the internal drainage group were significantly higher than those of the control group and the external drainage group (P ＜ 0.05).The expression of STAT3 expression of the internal drainage group reached peak at postoperative hour 4,which was 42.6％ ± 3.6％ ;the expression of STAT3 expression of the control group and the external drainage group reached peak at postoperative hour 12,which were 22.9％ ± 2.0％ and 29.2％± 3.7％.The peak level of STAT3 of the internal drainage group was significantly higher than those of the control group and the external drainage group (P ＜0.05).The levels of TNF-α and IL-6 of the internal drainage group reached peak at postoperative hour 12,which were (227 ±23)U/L and (256 ± 32)U/L; the levels of TNF-α and IL-6 of the control group and the external drainage group reached peak at postoperative hour 24,which were (309 ± 41) U/L and (388 ± 40) U/L,(287 ± 30)U/L and (346± 33)U/L,respectively.The levels of TNF-α and IL-6 of the internal drainage group at postoperative hour 0,1,2,4,12,24,48,72 were significantly lower than those of the control group and the external drainage group (P ＜ 0.05).The expressions of TNF-α mRNA of the control group,internal drainage group and external drainage group reached peak at postoperative hour 4,which were 0.92 ±0.14,0.39 ±0.05,0.80 ±0.15,respectively.The expressions of IL-6 mRNA reached peak at postoperative hour 12,which were 0.79 ± 0.07,0.38 ± 0.06,0.63 ±0.10,respectively.The expressions of TNF-α mRNA and IL-6 mRNA of the internal drainage group at postoperative hour 0,1,2,4,12,24,48,72 were significantly lower than those of the control group and the external drainage group (P ＜ 0.05).Conclusions Both internal and external drainage can improve liver regeneration of obstructive jaundice rats following partial hepatectomy,while the effect of internal drainage is superior.Internal biliary drainage has influence on the expression of STAT3 by decreasing the levels of TNF-α and IL-6,and help to improve liver regeneration of obstructive jaundice rats following partial hepatectomy.
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Objective To investigate the correlation between single nucleotide polymorphisms (SNPs) of FOXP3 gene and the susceptibility of hepatocellular carcinoma (HCC). Methods Two SNPs rs2280883 and rs3761549 of FOXP3 gene in 392 HCC patients and 372 healthy controls were analyzed by Matrix-Assisted Laser Desorption/ Ionization Time of Flight Mass Spectrometry (MALDI-TOF).Results At rs3761549,C allele frequency was significantly higher ( OR =1.32,95％ CI 1.03 -1.70,P =0.027) in HCC patients than healthy controls.Compared with healthy controls,HCC patients had higher frequencies of TT genotype (79.6％ ) at rs2280883 or CC genotype (77.6％) at rs3761549 of FOXP3 gene.Patients carrying rs2280883 TT genotype ( OR =1.53,95％ CI 1.10 - 2.14,P ＜ 0.00001 ) or rs3761549 CC genotype ( OR =1.92,95％ CI 1.39 - 2.64,P ＜ 0.00001 ) were more susceptible to HCC.Stratified analysis showed that rs3761549 CC genotype was significantly associated with higher incidence of portal vein tumor thrombus ( x2 =5.578,P =0.018 ),and rs3761549 TT/CT genotype was significantly associated with higher rate of tumor recurrence in HCC patients (x2 =6.561,P =0.010).Conclusions FOXP3 gene polymorphisms at rs2280883 and rs3761549 might be associated with increased susceptibility to HCC. rs3761549,CC genotype and TT/CT genotype were respectively associated with higher incidence of portal vein tumor thrombus and tumor recurrence in HCC patients.