Decrease in GPSM2 mediated by the natural product luteolin contributes to colon adenocarcinoma treatment and increases the sensitivity to fluorouracil.

Luteolin, a monomeric substance, is a natural product of the Brucea javanica (BJ) plant. Brucea javanica oil emulsion injection (BJOEI) is a proprietary Chinese medicine purified from BJ that is widely used clinically as an anti-tumor treatment. Although a growing body of research suggests that luteolin and BJOEI have anti-tumor effects, the molecular mechanism of action has not been fully elucidated. In this study, through molecular docking technology, we found that luteolin can interact directly with GPSM2 and regulate the FoxO signaling pathway through GPSM2. In addition, the inhibitory effect of luteolin on colon adenocarcinoma (COAD) cells was found to be offset by knockdown of GPSM2. In contrast, the anti-proliferative effects of luteolin could be notably reversed by overexpression of GPSM2. The results reveal that GPSM2 is crucial in luteolin-mediated anti-proliferative effects. The mediation of anti-proliferative effects by GPSM2 has also been indirectly demonstrated in RKO and SW480 xenograft mice models. In addition, we verified that BJOEI inhibits the progression of COAD by mediating GPSM2 and regulating the FoxO signaling pathway. We also found that BJOEI achieved a better anti-tumor effect when combined with fluorouracil injection. Collectively, our data show that the anti-tumor effects of BJOEI and luteolin on COAD are GPSM2-dependent and downregulating the expression of GPSM2 to regulate the FoxO signaling pathway may be an effective way to treat COAD.

The induction of SHP-1 degradation by TAOK3 ensures the responsiveness of T cells to TCR stimulation.

Thousand-and-one-amino acid kinase 3 (TAOK3) is a serine and threonine kinase that belongs to the STE-20 family of kinases. Its absence reduces T cell receptor (TCR) signaling and increases the interaction of the tyrosine phosphatase SHP-1, a major negative regulator of proximal TCR signaling, with the kinase LCK, a component of the core TCR signaling complex. Here, we used mouse models and human cell lines to investigate the mechanism by which TAOK3 limits the interaction of SHP-1 with LCK. The loss of TAOK3 decreased the survival of naïve CD4+ T cells by dampening the transmission of tonic and ligand-dependent TCR signaling. In mouse T cells, Taok3 promoted the secretion of interleukin-2 (IL-2) in response to TCR activation in a manner that depended on Taok3 gene dosage and on Taok3 kinase activity. TCR desensitization in Taok3-/- T cells was caused by an increased abundance of Shp-1, and pharmacological inhibition of Shp-1 rescued the activation potential of these T cells. TAOK3 phosphorylated threonine-394 in the phosphatase domain of SHP-1, which promoted its ubiquitylation and proteasomal degradation. The loss of TAOK3 had no effect on the abundance of SHP-2, which lacks a residue corresponding to SHP-1 threonine-394. Modulation of SHP-1 abundance by TAOK3 thus serves as a rheostat for TCR signaling and determines the activation threshold of T lymphocytes.

Microarray profiling of LncRNA expression in the testis of pubertal mice following morning and evening exposure to 1800 MHz radiofrequency fields.

In this paper, the chronotoxicity of radiofrequency fields (RF) in the pubertal testis development and the involved molecular pathways were investigated by exposing four-week-old mice to RF (1800 MHz, SAR, 0.50 W/kg) in the morning and evening of each day for three weeks. Then, pathological changes and functional indices within the testis were determined. We also used a long non-coding RNA (lncRNA) microarray and GO/KEGG pathway analyses to determine lncRNA expression profiles and predict their potential functions. The cis and trans regulation of lncRNAs were investigated, and an interaction network was constructed using Cytoscape software. RF exposure led to a range of pathological changes in the testes of adolescent mice, as testicular weights and daily sperm productions decreased, and the testosterone secretion reduced. Furthermore, RF induced dysregulation in the expression of testicular lncRNAs. We identified 615 and 183 differentially expressed lncRNAs that were associated with morning and evening exposure to RF, respectively. From 15 differential expression lncRNAs both in morning RF group and evening RF group, we selected 6 lncRNAs to be validated by quantitative reverse transcription PCR (qRT-PCR). The differentially expressed lncRNAs induced by morning RF exposure were highly correlated with many different pathways, including Fanconi syndrome, metabolic processes, cell cycle, DNA damage, and DNA replication. Trans-regulation analyses further showed that differentially expressed lncRNAs were involved in multiple transcription factor-regulated pathways, such as TCFAP4, NFkB, HINFP, TFDP2, FoxN1, and PAX5. These transcription factors have all been shown to be involved in the modulation of testis development, cell cycle progression, and spermatogenesis. These findings suggest that the extent to which 1800 MHz RF induced toxicity in the testes and changed the expression of lncRNAs showed differences between morning exposure and evening exposure. These data indicate that differentially expressed lncRNAs play crucial roles in the RF exposure damage to the developing pubertal testis. Collectively, our findings provide a better understanding of the mechanisms underlying the toxic effects of RF exposure on testicular development.

[Joint effects of nano-selenium and nano-cerium on the male reproductive function of mice exposed to microwave radiation].

OBJECTIVE: To study effect of nano-selenium and nano-cerium(nano cerium oxide) on the spermatogenic ability of mice irradiated by 1800 MHz microwave radiation(MR). METHODS: Forty-two ICR mice were randomly divided into groups: blank control group, solvent control group, microwave radiation model group, low, medium and high dose groups of nano-selenium+nano-cerium. In joint effects groups of nano-selenium and nano-cerium, the nano-selenium solution(60, 120 and 240 µg/kg) and the nano-cerium oxide solution(15, 30, 60 µg/kg) were administered to the stomach at 7:30 in the morning and 18:30 in the evening, respectively. The blank control group was orally administered with an equal volume of distilled water, and the solvent control group and the MR group were orally administered with an equal volume of carboxymethylcellulose sodium solution. During the second week of gastric administration, the mice were exposed to microwave radiation(1800 MHz) for 2 h every day(specific absorption ratio: 0. 2986 W/kg). After MR treatment, the daily sperm production of testis, sperm motility and sperm deformity rate in epididymis were measured, and the testicular marker enzymes [G6 PDH(6-phosphatedehydrogenase), ACP(acid phosphatase), LDH(lactate dehydrogenase)], antioxidant indexes [CAT(catalase), MDA(malondialdehyde) and T-AOC(total antioxidant capacity)] in testicular tissue were analyzed. RESULTS: Compared with the solvent control group, MR led to the decrease of sperm motility and the increase of sperm deformity rate, decreased the enzymes activities of G6 PDH, ACP and CAT, increased LDH activity and MDA content, and decreased the T-AOC level in testicular tissue, and the differences were statistically significant(P<0. 05). Compared with the MR group, the joint action of nano-selenium and nano-cerium with medium dose increased the daily sperm production of testis((18. 98±1. 27) ×10~6/g) vs. (15. 53±1. 24) ×10~6/g), decreased the sperm deformity rate(11. 74%±0. 91% vs. 16. 84%±2. 05%), and the joint action of nano-selenium and nano-cerium with medium and high dose increased the sperm motility in epididymis(61. 98%±6. 33%, 54. 17±4. 38 vs. 45. 16%±5. 01%), and the differences were statistically significant(P<0. 05). Compared with the MR group, the joint action of nano-selenium and nano-cerium with low and medium dose increased the activity of ACP(11. 07±0. 98, 14. 85±1. 39 vs. 8. 72±0. 91 nmol/(min·mg prot), P<0. 05). The joint action of nano-selenium and nano-cerium with medium and high dose increased the activity of G6 PDH(24. 12±2. 06, 21. 36±3. 65 vs. 15. 11±1. 73 nmol/(min·mg prot), P<0. 05) and decreased the activity of LDH(15. 52±1. 17, 13. 51±1. 68 vs. 22. 46±2. 01 nmol/(min·mg prot), P<0. 05). The joint action of nano-selenium and nano-cerium with medium dose increased the activity of CAT(17. 92±2. 03 vs. 11. 69±0. 87 nmol/(min·mg prot), P<0. 05) and decreased the content of MDA(5. 17 ±0. 62 vs. 9. 03 ±0. 63 nmol/mg prot, P<0. 05). The joint action of nano-selenium and nano-cerium with low, medium and high dose increased the level of T-AOC(22. 06±1. 54, 29. 36±2. 39, 21. 01±2. 47 vs. 12. 88±1. 82 U/mg prot, P<0. 05). CONCLUSION: The joint addition of nano-selenium and nano-cerium can improve the reproductive function of male mice exposed to MR, and can effectively alleviate the changes of mouse testicular marker enzyme activity and the decline of antioxidant capacity caused by MR.

MYSM1 maintains ribosomal protein gene expression in hematopoietic stem cells to prevent hematopoietic dysfunction.

Ribosomopathies are congenital disorders caused by mutations in the genes encoding ribosomal and other functionally related proteins. They are characterized by anemia, other hematopoietic and developmental abnormalities, and p53 activation. Ribosome assembly requires coordinated expression of many ribosomal protein (RP) genes; however, the regulation of RP gene expression, especially in hematopoietic stem cells (HSCs), remains poorly understood. MYSM1 is a transcriptional regulator essential for HSC function and hematopoiesis. We established that HSC dysfunction in Mysm1 deficiency is driven by p53; however, the mechanisms of p53 activation remained unclear. Here, we describe the transcriptome of Mysm1-deficient mouse HSCs and identify MYSM1 genome-wide DNA binding sites. We establish a direct role for MYSM1 in RP gene expression and show a reduction in protein synthesis in Mysm1-/- HSCs. Loss of p53 in mice fully rescues Mysm1-/- anemia phenotype but not RP gene expression, indicating that RP gene dysregulation is a direct outcome of Mysm1 deficiency and an upstream mediator of Mysm1-/- phenotypes through p53 activation. We characterize a patient with a homozygous nonsense MYSM1 gene variant, and we demonstrate reduced protein synthesis and increased p53 levels in patient hematopoietic cells. Our work provides insights into the specialized mechanisms regulating RP gene expression in HSCs and establishes a common etiology of MYSM1 deficiency and ribosomopathy syndromes.

CX-5461-loaded nucleolus-targeting nanoplatform for cancer therapy through induction of pro-death autophagy.

Various drugs have been designed in the past to act on intracellular targets. For the desired effects to be exerted, these drugs should reach and accumulate in specific subcellular organelles. CX-5461 represents a potent small-molecule inhibitor of rRNA synthesis that specifically inhibits the transcription driven by RNA polymerase (Pol) I and induces tumor cell death through triggering a pro-death autophagy. In the current study an innovative kind of CX-5461-loaded mesoporous silica nano-particles enveloped by polyethylene glycol (PEG), polydopamine (PDA) and AS-1411 aptamer (MSNs-CX-5461@PDA-PEG-APt) with the aim of treating cancer cells was constructed, in which the high-surface-area MSNs allowed for high drug loading, PDA acted as gatekeeper to prevent the leakage of CX-5461 from MSNs, PEG grafts on PDA surfaces increased the stable and biocompatible property in physiological condition, and AS-1411 aptamer promoted the nucleolar accumulation of CX-5461. MSNs-CX-5461@PDA-PEG-APt was characterized regarding releasing characteristics, steadiness, encapsulation of drugs, phase boundary potential as well as sizes of particles. Expectedly, In vitro assays showed that aptamer AS-1411 significantly increased the nucleolar accumulation of CX-5461. The aptamer-tagged CX-5461-loaded MSNs demonstrated to be more cytotoxic to cervical cancer cells compared to the control MSNs, due to relatively strong inhibition of rRNA transcription and induction of pro-death autophagy. The in vivo treatment with AS-1411-tagged CX-5461-loaded MSNs showed a stronger distribution in tumor tissues by animal imaging assay and a significantly higher inhibition effect on the growth of HeLa xenografts compared to AS-1411-untagged CX-5461-loaded MSNs. In addition, histology analysis indicated that MSNs-CX-5461@PDA-PEG-APt did not exhibit any significant toxicity on main organs. These results collectively suggested that MSNs-CX-5461@PDA-PEG-APt represents both a safe and potentially nucleolus-targeting anti-cancer drug. STATEMENT OF SIGNIFICANCE: Many drugs function in specific subcellular organelles. CX-5461 is a specific inhibitor of nucleolar rRNA synthesis. Here, we reported a novel aptamer-tagged nucleolus-targeting CX-5461-loaded nanoparticle, which specifically accumulated in nucleoli and significantly inhibited the tumor growth in vitro and in vivo through inhibiting rRNA transcription and triggering a pro-death autophagy.