IntroductionFollowing the implementation of the Heavy Metal Pollution Control Program in the Xiangjiang River Basin, significant progress has been achieved in the effective treatment and remediation of heavy metal contamination in the region. This study aims to identify and investigate microbial species that have adapted to long-term heavy metal contamination in soil environments, with a particular focus on their cadmium (Cd) tolerance mechanisms under stress conditions.MethodsUsing high-throughput Illumina HiSeq sequencing, differentially expressed genes were identified, annotated, and classified through comprehensive bioinformatics analyses. The associated signaling pathways were systematically examined in detail for functional interpretation.ResultsThis study highlights novel Cd-resistant genes exhibiting pronounced differential expression, thereby offering valuable insights into the Cd-tolerance mechanisms of filamentous fungi under heavy metal stress.DiscussionThese findings provide a solid foundation for improving fungal tolerance through targeted genetic modifications, such as small interfering RNA (siRNA) interference or gene overexpression, with potential applications in environmental bioremediation strategies.

Acute myeloid leukemia (AML) is maintained by a rare subpopulation of leukemic initiating cells, which drive disease progression, therapy resistance, and relapse. Transcription factor AP-2α, known for its role in tumor regulation, has an unclear function in leukemic initiating cells. Here, we found that low AP-2α expression correlated with poor AML prognosis, while high expression suppressed disease progression and improved survival. Functionally, AP-2α limited leukemic stem/initiating cell self-renewal and AML maintenance but was dispensable for normal hematopoiesis, as demonstrated using a Tfap2a conditional knockout model in hematopoietic stem cells. Moreover, loss of Tfap2a accelerated MLL-AF9 (MA9)-driven leukemogenesis in vivo, with knockout bone marrow exhibiting an expansion of leukemic granulocyte–monocyte progenitors, reflecting enhanced leukemia stem cell activity. Mechanistically, RNA-sequencing of Tfap2a-deficient cells revealed activation of inflammatory and stemness-related pathways, including JAK–STAT and TLR4 signaling. We further identified AP-2α as a direct transcriptional repressor of TLR4 by binding to its promoter, thereby exerting tumor-suppressive effects in AML-5 cells. Notably, the JAK2 inhibitor Fedratinib elevated AP-2α protein levels by attenuating STAT3 binding at the AP-2α promoter, positioning AP-2α as a downstream target of STAT3. Combining AP-2α overexpression with Fedratinib treatment or STAT3 knockdown synergistically inhibited AML-5 cell proliferation and leukemic stem cell self-renewal. These findings define a novel STAT3–AP-2α–TLR4 regulatory axis in AML and highlight AP-2α as a tumor suppressor that restrains leukemic stemness and disease progression. Therapeutic strategies that restore AP-2α expression or enhance its function—particularly in combination with JAK2 inhibition—may effectively target leukemic initiating cells in specific AML subtypes.

Hepatocellular carcinoma (HCC) is the predominant histologic subtype of primary liver cancer and accounts for approximately 90% of cases worldwide. Although immune checkpoint blockade (ICB) therapies targeting the PD-1/PD-L1 axis have demonstrated clinical promise in advanced HCC, therapeutic responses remain heterogeneous, underscoring the need to elucidate the mechanisms governing PD-L1 expression. Here, we identify potassium channel tetramerization domain-containing protein 1 (KCTD1) as a previously unrecognized regulator of PD-L1 in HCC. Mechanistically, KCTD1 enhances PD-L1 expression through stabilizing of the oncoprotein c-Myc. Immunofluorescence and co-immunoprecipitation assays reveal a direct interaction between KCTD1 and c-Myc, mediated by the BTB domain of KCTD1 and the BR-HLH-LZ domain of c-Myc. Knockdown of KCTD1 leads to decreased c-Myc and PD-L1 protein levels, concomitant with increased production of pro-inflammatory cytokines, including IFN-γ and TNF-α, and augmented CD8⁺ T cell cytotoxic activity in vitro. In a murine intrahepatic tumor model, KCTD1 knockdown synergizes with anti-PD-1 therapy, resulting in enhanced tumor infiltration by CD4⁺ and CD8⁺ T lymphocytes and improved anti-tumor efficacy. These findings establish KCTD1 as a key modulator of immune evasion in HCC and a promising target to potentiate immune checkpoint therapy.

The transcription factor AP-2α plays a crucial role in the control of tumor development and progression, and suppresses the proliferation and migration of hepatocellular carcinoma (HCC). However, the detailed function and mechanisms of AP-2α in the pathogenesis of HCC are still elusive. In the current study, we investigated the role of AP-2α regulation in liver injury-mediated HCC development. Downregulation of Tfap2a expression was found in the livers of DEN/CCl4-induced fibrosis and HCC mouse model. Hepatocyte (Alb-Cre), hepatic stellate cell (HSC) (Lrat-Cre) and macrophage (LysM-Cre) specific Tfap2a knockout mice were generated, respectively. Conditional knockout of Tfap2a was able to promote hepatic steatosis in Tfap2aΔHep and Tfap2aΔMΦ mice, but not in Tfap2aΔHSC mice fed with normal chow. Tfap2aΔHep and Tfap2aΔMΦ mice treated with DEN/CCl4 for 6 months increased tumor burden compared to Tfap2a flox controls. Tfap2a-deleted macrophages or hepatocytes could enhance lipid droplet (LD) accumulation in hepatocytes. Mechanistically, AP-2α binds to the promoter regions of SREBP1/ACC/FASN and inhibits hepatic lipid de novo synthesis. Deletion of Tfap2a in macrophages enhances polarization of M1 macrophages with increased iNOS expression but decreased CD206 expression, which resulted in increased pro-inflammatory cytokines and decreased anti-inflammatory factors, especially the hepatoprotective factor IL-10. The m6A modification writer WTAP could reduce the mRNA stability of AP-2α in a reader YTHDC1-dependent manner, whereas knockdown of WTAP or YTHDC1 enhances AP-2α expression and decreases lipid accumulation in HCC cells. Clinically, AP-2α expression negatively correlates with the expression of FASN, WTAP, YTHDC1 and the development of liver disease. Taken together, hepatocyte- or macrophage-specific deletion of Tfap2a promotes hepatic steatosis, fibrosis, and the development of HCC. These results suggest that AP-2α has been identified as a novel therapeutic target in fibrosis and inflammation-related HCC, exerting anti-lipogenesis, anti-inflammatory, and anti-tumor multi-roles.

Potassium channel tetramerization domain-containing 1 (KCTD1) plays a critical role in transcriptional regulation and adipogenesis, but its significance in hepatocellular cancer (HCC) has not been reported. Immunohistochemistry, Western blotting and quantitative real-time PCR analysis were performed to assess the expression of KCTD1 and related genes in HCC cells. MTT assays, colony formation, cell migration, invasion and the in-vivo mouse models were utilized to evaluate the function of KCTD1 in HCC progression. Co-immunoprecipitation, chromatin immunoprecipitation and luciferase reporter assays were conducted to elucidate the molecular mechanisms of KCTD1 in HCC. KCTD1 expression was increased in human HCC tissues and closely associated with advanced tumor stages. KCTD1 overexpression enhanced growth, migration, and invasion of Huh7 and HepG2 cells both in vitro and in vivo, while KCTD1 knockdown reversed these effects in MHCC97H cells. Mechanistically, KCTD1 interacted with hypoxia-inducible factor 1 alpha (HIF-1α) and enhanced HIF-1α protein stability with the inhibited prolyl-hydroxylases (PHD)/Von Hippel-Lindau (VHL) pathway, consequently activating the Vascular Endothelial Growth Factor (VEGF)/VEGFR2 pathway in HCC cells. Sorafenib and KCTD1 knockdown synergistically inhibited intrahepatic tumor growth following in situ injection of MHCC97H cells. miR-129-5p downregulated KCTD1 by binding to KCTD1 3′UTR. Finally, 45 µg exosomes from miR-129-5p-overexpressing MHCC97H cells combined with 25 mg/kg sorafenib to decrease HCC tumor size. These results suggested that KCTD1 protects HIF-1α from degradation and activates the VEGF signaling cascade to enhance HCC progression. Therefore, KCTD1 may serve as a novel target of HCC and pave the way for an efficient combined therapy in advanced HCC.

Lung cancer remains a critical global health concern, characterized by the highest incidence and mortality rates among all cancers. Due to its heterogeneity and complexity, the molecular mechanism underlying lung cancer occurrence and progression needs to be further investigated. KCTD10 has been implicated in malignant phenotypes of several tumors, but the role of KCTD10 in lung cancer remains largely unexplored. In this study, we found that KCTD10 expression is significantly reduced in lung cancer tissues, and overexpression of KCTD10 could inhibit lung cancer progression both in vitro and in vivo. Immunoprecipitation-mass spectrometry (IP-MS), co-immunoprecipitation (Co-IP), and ubiquitination assays revealed that the BTB domain of KCTD10 interacts with Armadillo repeat domains 1–9 of β-catenin and facilitates ubiquitin-dependent degradation of β-catenin via the K48-linked ubiquitin chains, followed by the downregulation of the β-catenin downstream target gene PD-L1. Notably, the combined treatment of KCTD10 overexpression with anti-PD-1 antibodies exhibited a synergistic effect in suppressing lung cancer progression and brain metastatic colonization in mice. In addition, vascular endothelial cell-specific knockout of Kctd10 (Kctd10flox/floxCDH5CreERT2/+) promoted lung cancer metastasis and tumor angiogenesis through β-catenin signaling. Finally, we identified METTL14- mediated N6-methyladenosine (m6A) modification within the coding sequence (CDS) region of KCTD10, which enhanced KCTD10 mRNA stability in a YTHDF2-dependent manner. These findings highlight KCTD10 as a critical regulator of lung cancer progression and the tumor microenvironment, suggesting its potential as a promising therapeutic target for lung cancer.

Anti-cancer peptides (ACPs) represent a promising potential for cancer treatment, although their mechanisms need to be further elucidated to improve their application in cancer therapy. Lycosin-I, a linear amphipathic peptide isolated from the venom of Lycosa singorensis, shows significant anticancer potential. Herein, it is found that Lycosin-I, which can self-assemble into a nanosphere structure, has a multimodal mechanism of action involving lipid binding for the selective and effective treatment of leukemia. Mechanistically, Lycosin-I selectively binds to the K562 cell membrane, likely due to its preferential interaction with negatively charged phosphatidylserine, and rapidly triggers membrane lysis, particularly at high concentrations. In addition, Lycosin-I induces apoptosis, cell cycle arrest in the G1 phase and ferroptosis in K562 cells by suppressing the PI3K-AKT-mTOR signaling pathway and activating cell autophagy at low concentrations. Furthermore, intraperitoneal injection of Lycosin-I inhibits tumor growth of K562 cells in a nude mouse xenograft model without causing side effects. Collectively, the multimodal effect of Lycosin-I can provide new insights into the mechanism of ACPs, and Lycosin-I, which is characterized by high potency and specificity, can be a promising lead for the development of anti-leukemia drugs.

Background: Eosinophils, a type of white blood cell originating from the bone marrow, are widely believed to play a crucial role in inflammatory processes, including allergic reactions and parasitic infections. However, the relationship between eosinophils and liver cancer is not well understood. Methods: Tumor immune infiltration scores were calculated using single-sample Gene Set Enrichment Analysis (ssGSEA). Key modules and hub genes associated with eosinophils were screened using Weighted LASSO regression, were used to identify prognostic genes and create a risk model. The Tumor Immune Dysfunction and Exclusion (TIDE) score was used to evaluate the immunotherapeutic significance of the analysis, real-time quantitative PCR (RT-qPCR), and Western blotting were used to determine gene expression levels and the status of eosinophil infiltration in tumors. Results: A risk trait model including 4 eosinophil-associated genes (RAMP3, G6PD, SSRP1, PLOD2) was developed by univariate Cox analysis and Lasso screening. Pathologic grading (p < 0.001) and model risk scores (p < 0.001) were found to be independent predictors of hepatocellular carcinoma (HCC) patient survival. Western blotting revealed higher levels of eosinophil peroxidase (EPX) in HCC tissues compared to adjacent normal tissues. Immunohistochemistry showed that eosinophils mainly infiltrated the connective tissue around HCC. The HCC samples showed low expression of RAMP3 and high expression of G6PD, SSRP1, and PLOD2, as detected by IHC and RT-qPCR analysis. The in vivo mouse experiments showed that IL-33 treatment induced the recruitment of eosinophils and reduced the number of intrahepatic tumor nodules. Conclusion: Overall, eosinophil infiltration in HCC is significantly correlated with patient survival. The risk assessment model based on eosinophil-related genes serves as a reliable clinical prognostic indicator and provides insights for precise treatment of HCC.

AIMS:The incidence of recurrent gliomas is high, exerting low survival rates and poor prognoses. Transcription factor AP-2α has been reported to regulate the progression of primary glioblastoma (GBM). However, the function of AP-2α in recurrent gliomas is largely unclear. METHODS:The expression of AP-2α and O6-methylguanine DNA-methyltransferase (MGMT) was detected in recurrent glioma tissues and cell lines by Western blots, the regulation mechanisms between AP-2α/MGMT promoter and RA/AP-2α promoter were studied by luciferase reporter assays, EMSA, and chIP assays. The effects of AP-2α and TMZ/RA treatment on cell viability in vitro and in vivo were investigated by MTT assays, γH2AX staining, comet assays and intracranial injection. KEY FINDINGS:AP-2α expression negatively correlates with the expression of MGMT in glioma samples. AP-2α could directly bind with the promoter of the MGMT gene, suppresses transcriptional levels of MGMT and downregulate MGMT expression in TMZ-resistant U87MG-R and T98G cells, but TMZ treatment decreases AP-2α expression and increases MGMT expression. The extended TMZ treatment and increased TMZ concentrations reversed these effects. Moreover, AP-2α overexpression combines with TMZ to decrease cell viability, concurrently with improved DNA damage marker γH2AX. Furthermore, retinoic acid (RA) activates RAR/RXR heterodimers, which bind to RA-responsive elements (RAREs) of the AP-2α promoter, and activates AP-2α expression in recurrent glioma cells. Finally, in intracranial relapsed glioma mouse model, both RA and TMZ could retard tumor development and prolong the mouse survival. SIGNIFICANCE:AP-2α activation by gene overexpression or RA treatment reveals the suppressive effects on glioma relapse, providing a novel therapeutic strategy against malignant refractory gliomas.

The coexistence of brown adipocytes with low and high thermogenic activity is a fundamental feature of brown adipose tissue heterogeneity and plasticity. However, the mechanisms that govern thermogenic adipocyte heterogeneity and its significance in obesity and metabolic disease remain poorly understood. Here we show that in male mice, a population of transcription factor jun-B (JunB)-enriched (JunB + ) adipocytes within the brown adipose tissue exhibits lower thermogenic capacity compared to high-thermogenic adipocytes. The JunB + adipocyte population expands in obesity. Depletion of JunB in adipocytes increases the fraction of adipocytes exhibiting high thermogenic capacity, leading to enhanced basal and cold-induced energy expenditure and protection against diet-induced obesity and insulin resistance. Mechanistically, JunB antagonizes the stimulatory effects of PPARγ coactivator-1α on high-thermogenic adipocyte formation by directly binding to the promoter of oestrogen-related receptor alpha, a PPARγ coactivator-1α downstream effector. Taken together, our study uncovers that JunB shapes thermogenic adipocyte heterogeneity, serving a critical role in maintaining systemic metabolic health.

程序性死亡受体1(PD-1)/程序性死亡配体1(PD-L1)信号通路主要参与免疫负调控作用,且在许多类型的肿瘤的恶性发展中具有关键作用.PD-L1的高表达可促进肝细胞癌(HCC)的侵袭,提高肿瘤复发的风险.另外,PD-L1常作为免疫检查点的阻断靶点,主要通过单抗将其中和,引发抗肿瘤免疫反应.因此,PD-L1是HCC免疫治疗中极具潜力的靶点之一.本文主要探究纳米级功能化氧化石墨烯(GO-PEI-PEG)携带PD-L1 siRNA对肝癌细胞的恶性生物学行为的影响.研究结果显示,将GO-PEI-PEG/PD-L1 siRNA转染至MHCC97H细胞后,细胞的增殖和迁移均被抑制,细胞周期阻滞在G1期,且细胞凋亡的数目增多.进一步研究发现,GO-PEI-PEG/PD-L1 siRNA对MHCC97H细胞的抑制作用是通过阻碍AKT信号通路激活实现的.这些试验结果表明,GO-PEI-PEG具备优秀的递送性能,携带PD-L1 siRNA可有效干扰PD-L1表达,进而抑制肝癌细胞的恶性生物学行为,这为治疗HCC提供了更安全、有效的递送新策略.

Programmed death-ligand 1 (PD-L1) ensures that tumor cells escape T-cell-mediated tumor immune surveillance. However, gliomas are characteristic of the low immune response and high-resistance therapy, it is necessary to understand molecular regulatory mechanisms in glioblastoma, especially the limited regulation of PD-L1 expression. Herein, we show that low expression of AP-2α is correlated with high expression of PD-L1 in high-grade glioma tissues. AP-2α binds directly to the promoter of the CD274 gene, not only inhibits the transcriptional activity of PD-L1 but enhances endocytosis and degradation of PD-L1 proteins. Overexpression of AP-2α in gliomas enhances CD8+ T cell-mediated proliferation, effector cytokine secretion, and cytotoxicity in vitro. Tfap2a could increase the cytotoxic effect of Cd8+ T cells in CT26, B16F10, and GL261 tumor-immune models, improve anti-tumor immunity, and promote the efficacy of anti-PD-1 therapy. Finally, the EZH2/H3K27Me3/DNMT1 complex mediates the methylation modification of AP-2α gene and maintains low expression of AP-2α in gliomas. 5-Aza-dC (Decitabine) treatment combines with anti-PD-1 immunotherapy to efficiently suppress the progression of GL261 gliomas. Overall, these data support a mechanism of epigenetic modification of AP-2α that contributes to tumor immune evasion, and reactivation of AP-2α synergizes with anti-PD-1 antibodies to increase antitumor efficacy, which may be a broadly applicable strategy in solid tumors.

The intermittent fasting (IF) diet has profound benefits for diabetes prevention. However, the precise mechanisms underlying IF's beneficial effects remain poorly defined. Here, we show that the expression levels of cyclooxygenase-2 (COX-2), an enzyme that produces prostaglandins, are suppressed in white adipose tissue (WAT) of obese humans. In addition, the expression of COX-2 in WAT is markedly upregulated by IF in obese mice. Adipocyte-specific depletion of COX-2 led to reduced fractions of CD4+Foxp3+ Tregs and a substantial decrease in the frequency of CD206+ macrophages, an increase in the abundance of γδT cells in WAT under normal chow diet conditions, and attenuation of IF-induced antiinflammatory and insulin-sensitizing effects, despite a similar antiobesity effect in obese mice. Mechanistically, adipocyte-derived prostaglandin E2 (PGE2) promoted Treg proliferation through the CaMKII pathway in vitro and rescued Treg populations in adipose tissue in COX-2-deficient mice. Ultimately, inactivation of Tregs by neutralizing anti-CD25 diminished IF-elicited antiinflammatory and insulin-sensitizing effects, and PGE2 restored the beneficial effects of IF in COX-2-KO mice. Collectively, our study reveals that adipocyte COX-2 is a key regulator of Treg proliferation and that adipocyte-derived PGE2 is essential for IF-elicited type 2 immune response and metabolic benefits.

Potassium channel tetramerization domain-containing 1 (KCTD1) plays a critical role in transcriptional regulation and adipogenesis, but the significance of KCTD1 in hepatocellular cancer (HCC) has not been reported. In this study, KCTD1 expression is increased in human HCC tissues and closely associated with advanced tumor stages. KCTD1 overexpression enhances HCC growth, migration, and invasion both in vitro and in vivo , while KCTD1 knockdown reverses these effects. Mechanistically, KCTD1 interacts with hypoxia-inducible factor 1 alpha (HIF-1α), enhances HIF-1α protein stability with the inhibited prolyl-hydroxylases (PHD)/Von Hippel-Lindau (VHL) pathway, consequently activating the Vascular Endothelial Growth Factor (VEGF)/VEGFR2 pathway in HCC cells. Sorafenib and KCTD1 knockdown synergistically inhibit intrahepatic tumor growth. miR-129-5p downregulates KCTD1 expression by binding with KCTD1 3′ UTR. Finally, exosomes derived from miR-129-5p-overexpressing HCC cells combine with sorafenib to decrease HCC tumor size. These results suggest that KCTD1 expression protects HIF-1α from degradation and activates the VEGF signaling cascade to enhance HCC progression. Therefore, KCTD1 may serve as a novel target of HCC and pave the way for an efficient combined therapy for advanced human HCC.Funding Information: This work was supported by the National Natural Science Foundation of China (No. 81872256), Key grant of research and development in Hunan Province (No. 2020DK2002), Natural Science Foundation in Hunan Province (NO.2021JJ30453), Key project of Hunan Provincial Education Department (No.19A310), Cooperative Innovation Center of Engineering and New Products for Developmental Biology of Hunan Province (No.20134486), as well as City University of Hong Kong Strategic Research Grant (No. 7005264).Declaration of Interests: These authors declare no competing financial interests.Ethics Approval Statement: This study was approved by the ethical principles and guidelines for Experiments on Animals and HCC tissues of Hunan Normal University (2018-035). Informed consent was gotten from all participants.

Programmed cell death receptor ligand 1 (PD-L1)/PD-1 signaling has been exploited to design inhibitors that deliver promising clinical outcome albeit with limited efficacy. Herein, we prepare graphene oxide (GO)-PEI-PEG with low cytotoxicity and long stability and GO-PEI-PEG delivers PD-L1 siRNAs to hepatocellular carcinoma (HCC) cells by the endocytosis-lysosome pathway. The functional GO-PEI-PEG/PD-L1 siRNAs decrease PD-L1 and PD-1 abundance, increase pro-inflammation cytokine IFN-γ and TNF-α release, and improve the proliferation activity of Jurkat T cells. Since GO-PEI-PEG targets the mouse liver effectively, the intrahepatic tumors in C57BL/6 mice are treated with GO-PEI-PEG/Pd-l1 siRNAs via the tail vein, resulting in shrinkage of the HCC tumors and boosting the anti-tumor efficacy in combination with oral sorafenib. A single treatment improves the total CD3+ and cytotoxic CD8+ T cell infiltration in the HCC tumor tissues and even spleen and upregulates the expression of Perforin, Gzmb, Ifng, Il-1b and Tnfa in the tumors after the combined treatment. Both the single and combined treatments enhance reactive oxygen species (ROS) accumulation, and improved HCC ferroptosis. The results suggest that GO-PEI-PEG delivered PD-L1 siRNAs combined with oral sorafenib can activate the adaptive immunity and tumor ferroptosis and reveal an effective therapy to treat advanced HCC patients.

肝癌是肝脏中最常见的恶性肿瘤,是全球癌症相关死亡的第三大主要原因.肝癌发病具有隐蔽性、细胞异质性、耐药性的特点,且肿瘤易侵袭、转移和复发,使得其临床治疗效果欠佳.转录因子AP-2α在肝癌中作为肿瘤抑制因子发挥作用,其表达与肝癌患者的预后呈现正相关.SOX9在细胞分化、性别决定和肿瘤发生中起主要作用.在肝癌细胞中,SOX9异常增加可以促进癌细胞的生长.本研究利用JASPAR软件预测到SOX9的启动子区域含有潜在的AP-2α结合位点,通过萤光素酶和凝胶迁移试验(EMSA)证实了AP-2α可以与SOX9的启动子区域直接结合,抑制SOX9的转录活性.通过实时定量PCR和蛋白质免疫印迹发现,AP-2α抑制了SOX9的mRNA和蛋白质水平表达.这些结果提示了AP-2α可与癌基因SOX9的启动子区域结合,负调控肝癌细胞中SOX9的表达.

为研究AP-2α在发育过程中的特定调控作用,利用AP-2α-LacZ转基因小鼠FX18研究AP-2α与先天性心脏病的相关性.在FX18小鼠繁殖过程中,发现该小鼠稳定遗传的突变体,鉴定其基因型,测量体质量与心脏质量,HE染色和心电图观察其表型和检测心脏功能.研究结果表明:突变体小鼠在出生后4周左右即死亡,且在胚胎期就表现出明显的发育迟缓,出生后的体质量和心脏质量均仅为正常FX18小鼠的40％～60％,同时,HE染色和心电图检测结果显示,小鼠表现出较明显的左心室肥大和室性早搏,符合先天性心脏病中动脉导管未闭的特征.因此,成功发现和建立的该突变体小鼠可作为一个较好的研究先天性心脏病小鼠模型.

Background: Potassium channel tetramerization domain-containing 1 (KCTD1) plays a critical role in transcriptional regulation and adipogenesis, but the significance of KCTD1 in hepatocellular cancer (HCC) has not been reported. Methods: KCTD1 expression was detected by immunohistochemistry, Western blotting, and quantitative RT-PCR. Malignant behaviors of cells were analyzed by MTT assays, liquid colony formation, scratch, transwell assays and FACS analysis. The in vivo functional assays were examined by mouse models using subcutaneous, tail vein, intrahepatic injection combined with sorafenib or exosome treatment. The angiogenesis was analyzed by tube formation and Matrigel plug model. The protein-protein interaction was demonstrated by Co-immunoprecipitation. The protein-DNA binding was confirmed by luciferase assays and chromatin immunoprecipitation. Results: KCTD1 expression is increased in human HCC tissues and closely associated with advanced tumor stages. KCTD1 overexpression enhances HCC growth, migration, and invasion both in vitro and in vivo , while KCTD1 knockdown reverses these effects. Moreover, KCTD1 overexpression promotes the in vitro growth, migration, and tube formation of human umbilical-vein endothelial cells (HUVECs) and induces in vivo angiogenesis. Mechanistically, KCTD1 interacts with HIF-1α to activate the VEGF pathway in HCC cells, which is concurrent with increased M2 macrophage infiltration. Sorafenib blocks the expression of KCTD1 protein and synergistically inhibits subcutaneous and intrahepatic tumor growth. miR-129-5p downregulates KCTD1 expression by binding with KCTD1 3′ UTR and abrogates the oncogenic role of KCTD1 in vitro . Finally, exosomes derived from miR-129-5p-overexpressing HCC cells combine with sorafenib to decrease HCC tumor size. Conclusions: These results suggest that KCTD1 expression activates the VEGF signaling cascade to enhance HCC progression, and angiogenesis. Therefore, KCTD1 may serve as a novel target of sorafenib and pave the way for an efficient therapy for advanced human HCC.

Curcumin, a phytochemical from rhizomes of the plant Curcuma longa, has been reported to exert potential anticancer properties in various cancer types, including acute myeloid leukemia (AML). However, the underlying mechanism remains poorly understood. The present study demonstrated that curcumin had a stronger cytotoxic activity against AML cells compared with three other types of phytochemicals (epigallocatechin gallate, genistein and resveratrol). Protein phosphorylation profiling using an antibody array identified that curcumin treatment increased the phosphorylation levels of 14 proteins and decreased those of four proteins. A protein-protein interaction network was constructed using the STRING database, in which AKT was identified as a hub protein with the highest connectivity (PRAS40, 4E-BP1, P70S6K, RAF-1 and p27). Western blotting results indicated that curcumin dose-dependently suppressed the phosphorylation of AKT, PRAS40, 4E-BP1, P70S6K, RAF-1 and p27 in AML cell lines (ML-2 and OCI-AML5). It was also demonstrated that curcumin regulated the cell cycle- and apoptosis-related proteins (cyclin D1, p21, Bcl2, cleaved-caspase-3 and cleaved-PARP), leading to cell cycle arrest and apoptosis in both ML-2 and OCI-AML5 cells. These effects of curcumin were enhanced by the AKT inhibitor afuresertib but were suppressed by the AKT activator SC-79, indicating that curcumin functions via AKT. In the AML xenograft mouse model, curcumin and afuresertib synergistically suppressed the engraftment, proliferation and survival of AML cells. Collectively, the present study demonstrated that curcumin exerted anti-AML roles by inactivating AKT and these findings may aid in the treatment of AML.