Aim: To study the antitumor mechanism of OSW-1 in hepatocellular carcinoma. Materials and Methods: The expression profiling microarray was carried out to extract RNA from SK-Hep-1 which suffered from OSW-1. ρ0-SK-Hep...Aim: To study the antitumor mechanism of OSW-1 in hepatocellular carcinoma. Materials and Methods: The expression profiling microarray was carried out to extract RNA from SK-Hep-1 which suffered from OSW-1. ρ0-SK-Hep-1 was maintained SK-Hep-1 in MEM containing 100 μg/L ethidium bromide (EB), 1 mM sodium pyruvate and 50 μg/ml uridine for 40 days. Then confirmed COX-I and COX-II of mitochondrial DNA were knocked out. Cells suffered from OSW-1 or doxorubicin. Then cells were washed twice with cold PBS and incubated with DCFH-DA. Fluorescent signal was recorded by using Infinite 200 Pro multimode Plate readers. Results: OSW-1 elevates generation of ROS and Cytochrome C which are associated with the induction of apoptosis in SK-Hep-1 cells. We also demonstrate that OSW-1 does not depend on p53 to up-regulate the BH3-only protein Noxa. What is more noteworthy that the Caspase-9 and FADD are down-regulated in above process. Conclusion: OSW-1 induced special apoptosis is different from the mitochondrial death pathway and the death receptor pathway and final result is not Caspase family’s activating. This provides a novel theory that nonmalignant cells are significantly less sensitive to OSW-1 than cancer cell lines.展开更多
To investigate the effect of mitochondrial DNA deletion and OSW-1 on PI3K-AKT signaling pathway PCR Array in SK-Hep1 hepatocellular carcinoma cells, we prepared SK-Hep1 cells with mtDNA deletion, that is, p0SK Hep. Th...To investigate the effect of mitochondrial DNA deletion and OSW-1 on PI3K-AKT signaling pathway PCR Array in SK-Hep1 hepatocellular carcinoma cells, we prepared SK-Hep1 cells with mtDNA deletion, that is, p0SK Hep. Then the OSW-1 of 100 ng/L was used to intervene SK-Hep1 and p0SK-Hep1. RT-qPCR was used to detect the difference of gene expression on PI3K-AKT signaling pathway PCR Array in four groups of cells. The gene expression of TLR4, FOS and TSC2 markers in SK-Hep1 cells treated with OSW-1 were significantly increased. The gene expressions of PDPK1, GJA1, TLR4 and TSC2 markers were significantly increased in p0SK-Hep1 cells, and the gene expressions of IRAK1 and GJA1 markers were significantly increased in p0SK-Hep1 cells treated with OSW-1. OSW-1 mainly affects the genes related to TLR4 pathway on PI3K-AKT signaling pathway PCR Array in SK-Hep1 HCC cells. P0SK-Hep1 mainly affects the upstream PDK1 gene and downstream TSC2 gene of Akt on PI3K-AKT signaling pathway PCR Array, and also affects the gene expression of gap junction at the same time.展开更多
5,6-Dihydro-OSW-1 (1) was synthesized following our previous procedure for the total synthesis of OSW-1. This compound demonstrated slightly stronger potency than that of OSW-1 against the growth of cancer cells.
文摘Aim: To study the antitumor mechanism of OSW-1 in hepatocellular carcinoma. Materials and Methods: The expression profiling microarray was carried out to extract RNA from SK-Hep-1 which suffered from OSW-1. ρ0-SK-Hep-1 was maintained SK-Hep-1 in MEM containing 100 μg/L ethidium bromide (EB), 1 mM sodium pyruvate and 50 μg/ml uridine for 40 days. Then confirmed COX-I and COX-II of mitochondrial DNA were knocked out. Cells suffered from OSW-1 or doxorubicin. Then cells were washed twice with cold PBS and incubated with DCFH-DA. Fluorescent signal was recorded by using Infinite 200 Pro multimode Plate readers. Results: OSW-1 elevates generation of ROS and Cytochrome C which are associated with the induction of apoptosis in SK-Hep-1 cells. We also demonstrate that OSW-1 does not depend on p53 to up-regulate the BH3-only protein Noxa. What is more noteworthy that the Caspase-9 and FADD are down-regulated in above process. Conclusion: OSW-1 induced special apoptosis is different from the mitochondrial death pathway and the death receptor pathway and final result is not Caspase family’s activating. This provides a novel theory that nonmalignant cells are significantly less sensitive to OSW-1 than cancer cell lines.
文摘To investigate the effect of mitochondrial DNA deletion and OSW-1 on PI3K-AKT signaling pathway PCR Array in SK-Hep1 hepatocellular carcinoma cells, we prepared SK-Hep1 cells with mtDNA deletion, that is, p0SK Hep. Then the OSW-1 of 100 ng/L was used to intervene SK-Hep1 and p0SK-Hep1. RT-qPCR was used to detect the difference of gene expression on PI3K-AKT signaling pathway PCR Array in four groups of cells. The gene expression of TLR4, FOS and TSC2 markers in SK-Hep1 cells treated with OSW-1 were significantly increased. The gene expressions of PDPK1, GJA1, TLR4 and TSC2 markers were significantly increased in p0SK-Hep1 cells, and the gene expressions of IRAK1 and GJA1 markers were significantly increased in p0SK-Hep1 cells treated with OSW-1. OSW-1 mainly affects the genes related to TLR4 pathway on PI3K-AKT signaling pathway PCR Array in SK-Hep1 HCC cells. P0SK-Hep1 mainly affects the upstream PDK1 gene and downstream TSC2 gene of Akt on PI3K-AKT signaling pathway PCR Array, and also affects the gene expression of gap junction at the same time.
基金the National Natural Science Foundation of China (Nos. 20372070 and 29925203).
文摘5,6-Dihydro-OSW-1 (1) was synthesized following our previous procedure for the total synthesis of OSW-1. This compound demonstrated slightly stronger potency than that of OSW-1 against the growth of cancer cells.
