Design, synthesis and activity evaluation of quinolinone derivatives as EZH2 inhibitors.

The enhancer of zeste homologue 2 (EZH2) is the core catalytic subunit of polycomb repressive complex 2, which catalyzes lysine 27 methylation of histone H3. Herein, a series of quinolinone derivatives were designed and synthesized based on the structure of Tazemetostat as the lead compound. Compound 9l (EZH2WT IC50 = 0.94 nM) showed stronger antiproliferative activity in HeLa cells than the lead compound. Moreover, compound 9e (EZH2WT IC50 = 1.01 nM) significantly inhibited the proliferation and induced apoptosis in A549 cells.

Design, synthesis and activity evaluation of Hedgehog inhibitor Itraconazole derivatives in A549 cells.

Hedgehog signal channel, a channel that plays a role in the occurrence and development of a variety of cancers. We introduce the composition and mechanism of Hedgehog signal channel, and the anti-tumor mechanism. A series of derivatives were designed and synthesized with the Hedgehog signal channel inhibitor Itraconazole in clinical phase II as the precursor. Compared with the other inhibitors, Itraconazole has a weaker inhibitory effect on Hedgehog, and there are few studies on improving the anti proliferation ability of Itraconazole on cells. Therefore, using Itraconazole as the lead, we obtained a series of derivatives that can effectively inhibit Hedgehog channels. Compared with Itraconazole, compounds 12g and 12n had stronger inhibitory effects in A549 cells.

Long non-coding RNA TPT1-AS1 alleviates cell injury and promotes the production of extracellular matrix by targeting the microRNA-324-5p/CDK16 axis in human dermal fibroblasts after thermal injury.

Long non-coding RNAs (lncRNAs) are associated with the healing of burn wounds in the dermis. The present study aimed to probe the role and regulatory network of the lncRNA TPT1 antisense RNA 1 (TPT1-AS1) in human dermal fibroblasts (HDFs) following thermal injury. A model of thermally injured cells was constructed with HDFs. The levels of TPT1-AS1, microRNA (miR)-324-5p and cyclin-dependent kinase (CDK)16 were determined through reverse transcription-quantitative PCR. Cell viability, cell cycle distribution, cell apoptosis rate and extracellular matrix (ECM) synthesis were assessed with a series of in vitro gain-of-function experiments and MTT, flow cytometry and western blot analyses. The binding ability of miR-324-5p and TPT1-AS1 (or the 3' untranslated region of CDK16) was identified via bioinformatics analysis and luciferase reporter assay. It was found that TPT1-AS1 and CDK16 were downregulated, but miR-324-5p was upregulated, in the HDFs after thermal injury. TPT1-AS1 elevation induced cell viability and ECM synthesis but attenuated cell cycle arrest at the G0/G1 stage and decreased the cell apoptosis rate of thermally injured HDFs. In addition, TPT1-AS1 sponged miR-324-5p to modulate CDK16 expression. Moreover, silencing CDK16 weakened the impacts of TPT1-AS1 upregulation on cell function and ECM synthesis in heat-treated HDFs. In summary, TPT1-AS1 relieved cell injury and induced ECM synthesis by sponging miR-324-5p and targeting CDK16 in the HDFs after thermal injury, implying a protective role for TPT1-AS1 in the burn wound healing process.

Silencing of long noncoding RNA SRRM2-AS exerts suppressive effects on angiogenesis in nasopharyngeal carcinoma via activating MYLK-mediated cGMP-PKG signaling pathway.

Long noncoding RNAs (lncRNAs) play a crucial role in several malignances, involving nasopharyngeal carcinoma (NPC), a heterogeneous disease. This study investigated mechanism of serine/arginine repetitive matrix protein 2-alternative splicing (SRRM2-AS) in NPC cell proliferation, differentiation, and angiogenesis. Initially, differentially expressed lncRNAs were screened out via microarray analysis. Vascular endothelial growth factor (VEGF) protein positive rate and microvessel density (MVD) were determined in NPC and adjacent tissues. NPC CNE-2 cells were treated with a series of vector and small interfering RNA to explore the effect of SRRM2-AS in NPC. The target relationship between myosin light chain kinase (MYLK) and SRRM2-AS was verified. Levels of SRRM2-AS, MYLK, cGMP, PKG, VEGF, PCNA, Ki-67, B-cell lymphoma-2 (Bcl-2), Bcl-2-associated X protein (Bax), and Caspase 3 were determined after transfection. Finally, the effect of SRRM2-AS on cell proliferation, colony formation, angiogenesis, cell cycle, and apoptosis in NPC was evaluated. SRRM2-AS was highly expressed and MYLK was poorly expressed in NPC tissues. VEGF protein positive rate and MVD were elevated in NPC tissues. MYLK was confirmed to be a target gene of SRRM2-AS. Silencing of SRRM2-AS elevated levels of MYLK, cGMP, PKG, Bax, and Caspase 3, but decreased levels of VEGF, PCNA, Ki-67, and Bcl-2. Especially, silencing of SRRM2-AS suppressed cell proliferation, colony formation and angiogenesis, blocked cell cycle, and enhanced cell apoptosis in NPC. Our results suggested that silencing of SRRM2-AS protected against angiogenesis of NPC cells by upregulating MYLK and activating the cGMP-PKG signaling pathway, which provides a new target for NPC treatment.

MicroRNA­146a/Toll­like receptor 4 signaling protects against severe burn­induced remote acute lung injury in rats via anti­inflammation.

The present study investigated the preventive effects of microRNA (miR)­146a against severe burn­induced remote acute lung injury (ALI) in rats and the underlying mechanism. The surface area of the skin was immersed in 100˚C water for 5­10 sec on the dorsal surface. The expression level of miR­146a was significantly downregulated in rats with burn­induced ALI. Downregulation of miR­146a increased inflammation, and inducible nitric oxide synthase (iNOS) and cyclooxygenase­2 (COX­2) expression in a model of ALI in vitro via the promotion of the Toll­like receptor (TLR)4/nuclear factor (NF)­κB signaling pathway. In addition, the overexpression of miR­146a reduced inflammation, and iNOS and COX­2 protein expression in the model of ALI in vitro via the suppression of the TLR4/NF­κB signaling pathway. A TLR4 inhibitor reduced the function of anti­miR­146a on inflammation in the model of ALI. Collectively, the results of the present study demonstrated the preventive effects of miR­146a against severe burn­induced remote ALI in rats through the anti­inflammatory­regulated TLR4/NF­κB signaling pathway.

Relationship between GSTM1/GSTT1 null genotypes and renal cell carcinoma risk: a meta-analysis.

The results from the published studies on the relationship between GSTM1/GSTT1 null genotypes and renal cell carcinoma (RCC) risk are still conflicting. This meta-analysis was performed to evaluate the relationship between GSTM1/GSTT1 null genotypes and RCC susceptibility. Association studies were identified from the databases of PubMed, Embase, Cochrane Library, and CBM-disc (China Biological Medicine Database) on 1 February 2012, and eligible investigations from 1950 to 2012 were synthesized using meta-analysis method. Results were expressed as odds ratios (ORs) for dichotomous data, and 95% confidence intervals (CIs) were also calculated. Six studies were identified for the analysis of association between polymorphic deletion of GSTM1/GSTT1 and RCC risk. There was no association between GSTM1/GSTT1 null genotype and RCC susceptibility (GSTM1: N = 6, p-heterogeneity = 0.07, OR = 1.07, 95% CI: 0.85-1.35, p = 0.57; GSTT1: N = 6, p-heterogeneity < 0.00001, OR = 0.98, 95% CI: 0.58-1.65, p = 0.94). Interestingly, null genotype of GSTT1 was associated with RCC risk in Caucasians and Asians (Caucasians: N = 4, p-heterogeneity = 0.38, OR = 0.76, 95% CI: 0.61-0.95, p = 0.01; Asians: N = 1, OR = 2.39, 95% CI: 1.63-3.51, p < 0.00001). For the GSTM1-GSTT1 interaction analysis, the dual null genotype of GSTM1/GSTT1 was not significantly associated with RCC susceptibility (N = 4, p-heterogeneity = 0.006, OR = 1.17, 95% CI: 0.98-1.39, p = 0.09). However, the dual null genotype of GSTM1-GSTT1 was associated with RCC risk in Asians (N = 1, OR = 2.06, 95% CI: 1.36-3.13, p = 0.007). In conclusion, our study results suggest that GSTT1 null genotype is associated with the RCC susceptibility in Caucasians and Asians, and the dual null genotype of GSTM1-GSTT1 is associated with RCC risk in Asians. However, more genetic epidemiological investigations are required to further explore this relationship.

[Experimental research on the effects of Chinese herb drugs on immune dysfunction in severely scalded rats].

OBJECTIVE: To observe the modulation effect of a concoction of Chinese herb drugs on immune dysfunction in severely burned rats. METHODS: One hundred healthy Wistar rats were randomly divided into A group (n = 30, with Chinese herb drug feeding after burns), B group (n = 30, with bouillon feeding after burns), C group (n = 30, with ordinary feeding after burns), and D group (n = 10, with sham burns as normal controls). The rats in A, B and C groups were subjected to 30% TBSA full-thickness burns and received conventional treatment. The rats in A group received 2 ml of Chinese herbal drug at 37 degrees C by gavage two times a day beginning from 2 postburn-hours(PBH). The rats in B group received 2 ml bouillon by gavage instead, and otherwise treatment was the same as A group, while the rats in C group were not fed by gavage. The activity of natural killer cell and T lymphocyte, and the levels of IgA, IgG, IgM, C3, C4 in A, B, C groups were examined on 3, 7, 14 PBD, and these indices were also determined in D group. RESULTS: Compared with D group, the amount of the CD3+, CD4+ lymphocyte, the ratio of the CD4+/CD8+, the level of IgA, IgG, IgM, C3, C4, the activity of NK cells, and the density of the sIgA in A, B, C groups were obviously decreased, but the amount of the CD8+ were obviously increased (P < 0.05 or P < 0.01). Furthermore, the above indices in A group improved more quickly when compared with B and C groups. CONCLUSION: The concoction of Chinese herb drugs can improve the distribution of T lymphocyte subsets, increase the activity of NK cells, promote the secretion of sIgA in intestinal mucous membrane and promote recovery of IgM, IgG, C3, C4 levels, thereby improves the immune function of the body.