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Objective To investigate the killing effect of ethacrynic acid (EA) on lung cancer A549 cells derived spheres and explore the underlying mechanism.Methods A549 spheres were cultured in serum-free medium,and the protein expression of CD133,SOX2,EpCAM and ABCG2 was detected by Western blotting.MTT assay was used to evaluate the cell viability of A549 spheres and A549 cells after treated by 1,2,5,10 and 20 mg/mL cisplatin (DDP) for 48 h.The activity of glutathione S-transferase (GST) was measured by colorimetric method after A549 spheres were treated with 10,50,100 and 200 μmol/L EA,respectively.Flow cytometry,Western blotting,real-time PCR and luciferase assay were used to analyze the levels of cellular reactive oxygen species (ROS),formation of A549 spheres,mRNA and protein expression levels of β-catenin,Sox2 and ABCG2,and promoter activity of β-catenin upon 200 μmol/L EA treated cells for 48 h.A549 sphere was infected with β-catenin adenovirus for 24 h,followed by 200 μmol/L EA treatment (in presence or absence of 5 mg/mL DDP) for 24 h.The expression of β-catenin,Sox2 and ABCG2 at mRNA and protein levels was detected by real-time PCR and Western blotting,and cell growth of A549 spheres was evaluated by MTT assay.Results The A549 spheres,with high expression of tumor stem cells markers CD133,SOX2,EpCAM and drug resistance related molecule ABCG2,and resistance to DDP at different doses,were successfully derived.After 200 μmol/L EA had treated A549 sphere for 48 h,the levels of ROS were significantly increased (P ＜ 0.05),and the mRNA and protein levels of β-catenin,Sox2 and ABCG2,and promoter activity of β-catenin were notably decreased (P ＜ 0.05).The treatment of 200 μmol/L EA enhanced the inhibitory effect on proliferation and the promoting effect on apoptosis in A549 spheres induced by 5 mg/mL DDP (P ＜ 0.05).Up-regulation of β-catenin by adenoviral infection partly reversed the effects of 200 μmol/L EA on suppressing the expression levels of β-catenin,Sox2 and ABCG2,compared to the spheres infected with blank adenovirus.Additionally,β-catenin over-expression significantly remitted the inhibitory effect of 200 μmol/L EA and 5 mg/mL DDP on the proliferation in A549 spheres.Conclusion EA exerts inhibitory effect on the proliferation and stemness of A549 spheres through suppressing GST activity and β-catenin expression,and then promotes cell apoptosis.EA might be a novel drug in treatment of lung cancer and cancer stem cells.
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<p><b>OBJECTIVE</b>To investigate the effect of low and high oxygen concentration on embryo development, pregnancy outcome and birth defects offertilization-embryo transfer (IVF-ET).</p><p><b>METHODS</b>According to the oxygen concentration ofculture environment, the IVF-ET performed in the Women's Hospital, Zhejiang University School of Medicine during 2013 and 2015 were divided into low oxygen concentration group (=2036, 5% O) and high oxygen concentration group (=4617, 20% O). The rate of fertilization, good quality embryo, clinical pregnancy, ectopic pregnancy, abortion and birth defect were compared between two groups.</p><p><b>RESULTS</b>The good quality embryo rate was significantly higher in the low oxygen concentration group (<0.05). However, no significant differences were found between two groups in the rate of fertilization, clinical pregnancy, ectopic pregnancy, abortion and birth defect (all>0.05).</p><p><b>CONCLUSIONS</b>Low oxygen environment may improve the potential of embryonic development, but its impact on pregnancy outcome and birth defect is not significant.</p>
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<p><b>OBJECTIVE</b>To investigate the effects of embryo cryopreservation and thawing on clinical outcomes of transplantable embryos after preimplantation genetic diagnosis (PGD) or preimplantation genetic screening (PGS) in cleavage-stage.</p><p><b>METHODS</b>The clinical data of 302 cases (including 118 cases using frozen/thawing embryos and 184 cases using fresh embryos) undergoing PGD/PGS in Women's Hospital, Zhejiang University School of Medicine during January 2011 and December 2016 were retrospectively analyzed. The pregnancy rate, implantation rate, live birth rate and abortion rate of fresh and frozen-thawed embryo transfer (FET) cycles were compared. And the influencing factors for pregnancy outcome was analyzed by multivariate logistic regression.</p><p><b>RESULTS</b>The rate of normal or balanced translocation embryos in fresh cycle was higher than that in FET cycle (23.52% vs 16.67%,<0.05), and the average number of transplanted embryos was more than that in FET cycle (1.54±0.56 vs 1.33±0.51,<0.05). But there were no significant differences in pregnancy rate (36.42% vs 40.00%,>0.05), implantation rate (26.62% vs 32.91%,>0.05), abortion rate (19.44% vs 8.33%,>0.05) and live birth rate (25.96% vs 28.33%,>0.05) between fresh cycle and FET cycle. Multivariate logistic regression showed that, parent ages, embryo status (fresh or frozen), the mode of PGD/PGS and the findings of PGD/PGS had no impact on pregnancy outcome (all>0.05).</p><p><b>CONCLUSIONS</b>Cryopreservation do not have significant effects on the clinical outcomes of transplantable embryos after PGD/PGS in cleavage-stage.</p>
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Hippo pathway is a signaling network involved in the regulation of cell proliferation and apoptosis, whereas Yes-asso-ciated protein (YAP) is the major effector of the pathway. YAP is a candidate oncogene, and dysregulation of the Hippo-YAP pathway is closely related to the occurrence and progression of various tumors. Therefore, Hippo-YAP may provide a novel therapeutic target for cancer treatment. Some drugs or compounds that regulate the Hippo-YAP pathway activity, such as verteporfin, can inhibit the occur-rence or growth of tumors. This review mainly focuses on the progress of studies on the use of Hippo-YAP signaling pathway as a tar-get of cancer treatment strategies.
ABSTRACT
Hypoxic microenvironment is a typical characteristic of solid tumors and is considered an independent risk factor in tu-mor progression and poor prognosis. Hypoxic microenvironment is a critical part of cancer stem cell (CSC) niche and plays an impor-tant role in the evolution of cancer stem cells in tumors and in apoptosis resistance. As a key factor in the tumor's adaption to hypoxic microenvironment, hypoxia inducible factors (HIFs) can induce biological behavioral changes in CSCs that can accelerate tumor malig-nant transformation. Such behavioral changes include anti-apoptosis, enhancement of drug-resistance gene expression, and tumor inva-sion and metastasis. HIFs are also among the main factors affecting the capacity of CSCs to maintain their biological characteristics. In this review, the authors focus on recent advances in our understanding of the role that HIFs play in maintaining the biological character-istics of CSCs.