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Trametinib is a MEK1/2 inhibitor and exerts anticancer activity against a variety of cancers. However, the effect of Trametinib on colorectal cancer (CRC) is not well understood. In the current study, our results demonstrate the ability of sub-toxic doses of Trametinib to enhance TRAIL-mediated apoptosis in CRC cells. Our findings also indicate that Trametinib and TRAIL activate caspase-dependent apoptosis in CRC cells. Moreover, Mcl-1 overexpression can reduce apoptosis in CRC cells treated with Trametinib with or without TRAIL. We further demonstrate that Trametinib degrades Mcl-1 through the proteasome pathway. In addition, GSK-3β phosphorylates Mcl-1 at S159 and promotes Mcl-1 degradation. The E3 ligase FBW7, known to polyubiquitinate Mcl-1, is involved in Trametinib-induced Mcl-1 degradation. Taken together, these results provide the first evidence that Trametinib enhances TRAIL-mediated apoptosis through FBW7-dependent Mcl-1 ubiquitination and degradation., (© 2020 The Authors. Journal of Cellular and Molecular Medicine published by Foundation for Cellular and Molecular Medicine and John Wiley & Sons Ltd.)

Apoptosis of vascular smooth muscle cells (VSMCs) is a process that regulates vessel remodeling in various cardiovascular diseases. The specific mechanisms that control VSMC apoptosis remain unclear. The present study aimed to investigate whether microRNA‑494 (miR‑494) is involved in regulating VSMC apoptosis and its underlying mechanisms. Cell death ELISA and terminal deoxynucleotidyl‑transferase‑mediated dUTP nick end labeling assays were used to detect apoptosis of murine VSMCs following stimulation with tumor necrosis factor‑α (TNF‑α). The results indicated that TNF‑α upregulated VSMC apoptosis in a dose‑dependent manner. Microarray analysis was used to evaluate the expression profile of microRNAs following TNF‑α stimulation in murine VSMCs. The expression of miR‑494 was downregulated, whereas B‑cell lymphoma-2‑like 11 (BCL2L11) protein expression levels were upregulated in VSMCs following treatment with TNF‑α. Luciferase reporter assays confirmed that BCL2L11 was a direct target of miR‑494. Transfection with miR‑494 mimics decreased VSMC apoptosis and downregulated BCL2L11 protein levels. Conversely, transfection with miR‑494 inhibitors increased cell apoptosis and upregulated BCL2L11 protein levels, suggesting that miR‑494 may function as an essential regulator of BCL2L11. The increase in apoptosis caused by miR‑494 inhibitors was abolished in cells co‑transfected with BCL2L11‑targeting small interfering RNA. The findings of the present study revealed that miR‑494 inhibited TNF‑α‑induced VSMC apoptosis by downregulating the expression of BCL2L11.

The p53 protein is one of the best-known tumor suppressors. Recently discovered ASPP1 and ASPP2 are specific activators and iASPP is an inhibitor of p53. In the present study, we found that of 37 NSCLC patients, p53 alterations were detected in 20 tumors (54.1%), the mRNA expression of ASPP1 and ASPP2 was frequently dowregulated in tumor tissues, and this decreased significantly in samples expressing wild-type p53. The expression of ASPP1 and ASPP2 was downregulated and that of iASPP was upregulated in two NSCLC cell lines (the NCI-H157 cell line with altered p53 and the A549 cell line with wild-type p53). The NCI-H157 cell with higher ASPP1 and ASPP2 levels was more sensitive to cisplatin than the A549 cells with lower ASPP1 and ASPP2 levels. Downregulation of iASPP by siRNA stimulated apoptosis through p53 in two NSCLC cell lines. These findings provide new insights into the molecular mechanisms of action of the ASPP family in NSCLC and may have potent therapeutic applications.

Protective effects of liraglutide on H9C2 cells cultured using CoCl2 and its mechanism of action were investigated. Hypoxia model was established using CoCl2-treated H9C2 cells. With liraglutide as the treatment factor, apoptosis, changes in nitric oxide (NO) and reactive oxygen species (ROS) activity, mitochondrial membrane potential and change in cell autophagy level were detected via Hoechst staining, enzyme-linked immunosorbent assay (ELISA), polymerase chain reaction (PCR) and western blotting (WB), respectively. Liraglutide ameliorated the CoCl2-induced decrease in H9C2 cell viability, the increases in cytotoxicity and percentage of apoptotic cells as well as oxidative stress in cells. Moreover, it stimulated the elevation of cell autophagy level. However, the protective effects of liraglutide on H9C2 cells were attenuated remarkably after adding the cell autophagy inhibitor. Liraglutide can ameliorate the CoCl2-induced oxidative stress and apoptosis in H9C2 cells via regulating cell autophagy. [ABSTRACT FROM AUTHOR]

Tetrachlorobisphenol A (TCBPA), a halogenated flame retardant and endocrine disruptor, has been detected in human urine and serum. While previous research has shown its impact on the reproductive system, investigations into its mechanisms during puberty remain limited. This study aims to explore the effects of TCBPA on Leydig cells in adolescent mice and potential underlying mechanisms. Male C57 mice of age 28 days were gavaged with 50, 100, and 200 mg/kg/day for 28 days. TCBPA did not alter body weight and testis weight but lowered testosterone levels at 100 and 200 mg/kg and reduced sperm count in the epididymis at 200 mg/kg. TCBPA lowered Leydig cell number at 200 mg/kg while it downregulated key Leydig cell gene (Lhcgr, Scarb1, Cyp11a1, Cyp17a1, Hsd3b6, Hsd17b3 and Insl3) as low as 50 mg/kg. Further study indicated that TCBPA induced reactive oxygen species and caused endoplasmic reticulum stress. In vitro study in TM3 mouse Leydig cells showed that TCBPA indeed induced reactive oxygen species and caused endoplasmic reticulum stress at 75 μM and inhibited testosterone production at this concentration and addition of antioxidant tocopherol can reverse it. These discoveries provide new insights and references for a deeper understanding of the toxic mechanisms of TCBPA on Leydig cells during puberty. [Display omitted] • Tetrachlorobisphenol A inhibits Leydig cell development in mice in puberty. • Tetrachlorobisphenol A reduces Leydig cell number in mice in puberty. • Tetrachlorobisphenol A induces ROS in mice in puberty. • Tetrachlorobisphenol A induces endoplasmic reticulum stress and apoptosis in TM3 Leydig cells. [ABSTRACT FROM AUTHOR]

Exposure to 4-nonyl phenol (4-NP) on Leydig cell (LC) development and function remains poorly understood. We explored the effects of 4-NP on LC development and elucidate the underlying mechanisms. Male (28-day-old) mice received orally 4-NP (0.125, 0.25, and 0.5 mg/kg/day) for 28 days. We found that 4-NP at ≥ 0.125 mg/kg markedly compromised serum testosterone levels and LC numbers. Gene and protein expression analysis demonstrated downregulation of key genes and their proteins involved in LC steroidogenesis, including Star, Cyp11a1, Cyp17a1, Hsd17b3, Hsd3b6, and Scarb1. Furthermore, exposure to 4-NP induced oxidative stress, as evidenced by elevated reactive oxygen species (ROS) and malondialdehyde (MDA), as well as reduced superoxide dismutase 1/2 and catalase (CAT). Apoptosis was also observed in LCs following exposure to 4-NP, as shown by an increased BAX/BCL2 ratio and caspase-3. A TM3 mouse LC line further confirmed that 4-NP induced ROS and the expression of apoptosis-related genes and proteins. In conclusion, this study demonstrates that 4-NP exposure compromises LC development through multiple mechanisms. [Display omitted] • 4-Nonylphenol inhibits Leydig cell development in mice in puberty. • 4-Nonylphenol reduces Leydig cell number in mice in puberty. • 4-Nonylphenol induces ROS in mice in puberty. • 4-Nonylphenol induces ROS and apoptosis in TM3 Leydig cells. [ABSTRACT FROM AUTHOR]

Chemotherapy resistance represents a major issue associated with gastric cancer (GC) treatment, and arises through multiple mechanisms, including modulation of the cell-cycle check point. Several ubiquitin kinases, including RING finger protein 138 (RNF138), have been reported to mediate the G2/M phase arrest. In this study, we investigated the role of RNF138 in the development of cisplatin resistance of two GC cell lines. We show that RNF138 levels are higher in cisplatin-resistant cell lines, compared with cisplatin-sensitive cells, and RNF138 expression was elevated during drug withdrawal following the cisplatin treatment. Using gene overexpression and silencing, we analyzed the impact of altering RNF138 level on GC cell viability, apoptosis, and cell cycle phenotypes in two isogenic cisplatin-sensitive and resistant cell lines. We show that RNF138 overexpression increased GC cell viability, decreased apoptosis and delayed cell cycle progression in the cisplatin-sensitive GC cells. Conversely, RNF138 silencing produced opposite phenotypes in the cisplatin-resistant cells. Moreover, RNF138-dependent phosphorylation of Chk1 was seen in GC cells, indicating a novel connection between cisplatin-induced DNA damage and apoptosis. Collectively, these data suggest that RNF138 modulates the cisplatin resistance in the GC cells, thus serving as a potential drug target to challenge chemotherapy failure. In addition, RNF138 can also be used as a marker to monitor the development of cisplatin resistance in GC treatment. [ABSTRACT FROM AUTHOR]

Background: Ras association domain family 1A gene (RASSFlA) is a candidate suppressor gene, Lack of RASSF1A expression was found in lung cancer. High DNA methylation at the promoter region is the main reason for inactivating RASSF1A transcription. Methods: In this study, we examined RASSF1A's methylation status and its mRNA expression level between non-small cell lung cancer cell line A549 and anti-Cisplatin cell strain A549DDP, Furthermore, methylation of A549DDP was reversed by treatment of 5-Aza-2' - deoxycytidine (5-Aza-cdR),a DNA methyltransferase inhibitor. Results: We found that RASSF1A's methylation status and its mRNA expression were obvious differences between A549 and A549DDP. 5-Aza-CdR treatment remarkablly reduced cell vability of A549DDP. Moreover, 5-Aza-CdR treatment induced A549DDP cell apoptosis in a dose dependent manner with declining cell percentage in S and G2/M stage, and increasing proportion in G0/G1 stage. Cell motility was blocked in G0/G1 stage. All of A549DDP cells showed unmethylated expression, its high methylation status was reversed in a dose-dependent manner within a certain range. Conclusions: The abnormal gene methylation status of RASSF1A is a molecular biomarker in lung cancer diagnosis, treatment and prognosis. [ABSTRACT FROM AUTHOR]

Background: The significance of CXCL5 in pancreatic cancer is unclear, although it has been implicated in the malignant process of many different types of cancer. Research on the impact of CXCL5 on immune cell infiltration and the malignant phenotype of pancreatic cancer is needed. This study aimed to examine the connection between CXCL5 expression and immune cell infiltration and the malignant phenotype of pancreatic cancer. Methods: Tissue samples and clinical information were collected from 90 patients with pancreatic cancer. Tumour tissues and adjacent tissues were made into a tissue microarray and stained for immunohistochemistry analysis. Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and Western blot analysis were performed to measure the expression level of CXCL5. CXCL5-overexpressing/CXCL5-knockdown cell lines were constructed via transfection for cytological experiments. CCK-8, cell apoptosis, cell cycle, cell invasion, and cell colony formation assays were used to detect the effect of CXCL5 on the malignant phenotype of pancreatic cancer cells. Finally, a mouse model of pancreatic cancer was constructed for in vivo verification. Results: Compared with control cells, pancreatic cancer cells overexpressing CXCL5 exhibited increased proliferation, migration, and invasion but decreased apoptosis. Conversely, knockdown of CXCL5 did not enhance the malignant phenotype of pancreatic cancer cells. Spearman correlation analysis indicated that there was a significant negative correlation between CXCL5 levels and the CD8 IRS. However, there was a significant positive correlation between FOXP3 IRS and CXCL5 levels. Conclusions: CXCL5 is highly expressed in pancreatic cancer and promotes the malignant phenotype of pancreatic cancer cells. CXCL5 is associated with immunosuppressive FOXP3 + T-cell infiltration, which facilitates the formation of an immunosuppressive microenvironment (with low CD8 + T-cell infiltration). [ABSTRACT FROM AUTHOR]