• HNC
• SMAD2
• SMAD4
• diagnostic markers
• exosomal microRNA
• exosomes
• miR-17-92 polycistronic cluster
• miR-19a
• miR-19b
• quantitative PCR

• Humans
• Exosomes/genetics
• Exosomes/metabolism
• MicroRNAs/genetics
• MicroRNAs/blood
• Female
• Male
• Head and Neck Neoplasms/genetics
• Head and Neck Neoplasms/pathology
• Head and Neck Neoplasms/blood
• Head and Neck Neoplasms/diagnosis
• Middle Aged
• Biomarkers, Tumor/genetics
• Gene Expression Regulation, Neoplastic
• Prognosis
• Smad4 Protein/genetics
• Adult
• Case-Control Studies
• Smad2 Protein/genetics
• Aged
• Kaplan-Meier Estimate
• ROC Curve

• KDM6A
• KMT2
• KMT2C
• KMT2D
• PDAC

• Humans
• Carcinoma, Pancreatic Ductal/genetics
• Carcinoma, Pancreatic Ductal/metabolism
• Carcinoma, Pancreatic Ductal/pathology
• Pancreatic Neoplasms/genetics
• Pancreatic Neoplasms/metabolism
• Pancreatic Neoplasms/pathology
• Epigenesis, Genetic
• Gene Expression Regulation, Neoplastic
• Chromatin/metabolism
• Chromatin/genetics
• DNA-Binding Proteins/metabolism
• DNA-Binding Proteins/genetics
• Histone Demethylases/metabolism
• Histone Demethylases/genetics
• Animals

• R37 CA262209 NCI NIH HHS
• 5R37CA262209-02A1 NIH HHS

• EMT
• cancer stem-like cells
• esophageal squamous cell carcinoma
• microenvironment
• surface markers

• MLL2
• epithelial-mesenchymal transition
• metastasis
• miR-147b
• tumor suppressor

• Humans
• Animals
• Mice
• Cell Plasticity
• Cell Line, Tumor
• MicroRNAs/genetics
• MicroRNAs/metabolism
• Pancreatic Neoplasms/pathology
• Transforming Growth Factor beta/metabolism
• Activins/genetics

• K08 CA234222 NCI NIH HHS
• R37 CA262209 NCI NIH HHS

• cell-free DNA
• distant metastasis
• liquid biopsy
• mutation burden
• oral cancer
• whole-exome sequencing

• 108-2314-B-195 -002 -MY2 National Science and Technology Council (NSTC), Taiwan
• 110-2314-B-195 -015 -MY3 National Science and Technology Council (NSTC), Taiwan
• MMH-E-105-12 and MMH-E-108-12 Mackay Memorial Hospital

• ESCC
• Epigenetic alterations
• Genetic variation
• Oncogene
• Signaling pathway

• CRISPR/Cas9
• Cancer
• Delivery
• In Vivo Models
• Mutations
• Validation

• CRISPR-Cas Systems
• Gene Editing
• Humans
• Mutation
• Neoplasms/genetics
• Oncogenes
• RNA, Guide, CRISPR-Cas Systems/genetics

Epigenetic reprogramming is involved in multiple steps of human cancer evolution and is mediated by a variety of chromatin-modifying enzymes. Specifically, the histone lysine methyltransferase KMT2D is among the most frequently mutated genes in oral squamous cell carcinoma (OSCC). However, the mechanisms by which KMT2D affects the development of OSCC remain unclear.

In the present study, we found that the expression of KMT2D was elevated in OSCC compared to paracancerous specimens and was correlated with a more advanced tumor grade. More importantly, knockdown of KMT2D impaired their reconstitution in patient-derived organoids and decreased the expression of CD133 and β-catenin in OSCC cells. In in vitro and in vivo models, knockdown of KMT2D reduced the colony formation, migration and invasion abilities of OSCC cells and delayed tumor growth. Mechanistically, the dual-luciferase reporter and co-immunoprecipitation assays in two individual OSCC cell lines indicated that KMT2D may cooperate with MEF2A to promote the transcription activity of CTNNB1, thereby enhancing WNT signaling.

The upregulation of KMT2D contributes to stem-like properties in OSCC cells by sustaining the MEF2A-mediated transcriptional activity of CTNNB1.

The online version contains supplementary material available at 10.1186/s13578-022-00785-8.

• CTNNB1
• KMT2D
• MEF2A
• OSCC
• Organoid
• Stemness

• Histone-Lysine N-Methyltransferase/genetics
• Histone-Lysine N-Methyltransferase/metabolism
• Histones/genetics
• Humans
• Mutation
• Neoplasms/genetics
• Prognosis

• 81372332, 81572822 and 81972908 National Natural Science Foundation of China
• WKJ-ZJ-2117 National Health Commission Science Research Fund-Zhejiang Provincial Health Key Science and Technology Plan Project
• LZ13H160002 and LZ18H160002 Key Projects from Zhejiang Provincial Natural Science Foundation
• LQ19H160005 and LY17H160044 General Project from Zhejiang Provincial Natural Science Foundation
• Zhejiang Provincial CPC Committee Talents [2019]-3 Leading Talents in Scientific and Technological Innovation from Zhejiang Provincial Ten Thousand Talents Plan
• Zjwjw2014-108 Zhejiang Provincial Program for Cultivation of High-Level Innovative Health Talents
• Zjhrss2014-150 Major Training Personnel from Zhejiang Provincial Program for Training and Development Project for 151 Talents
• 20IRTSTHN026 Innovation Research Team (in Science and Technology) in University of Henan Province
• 2019ZD025 Zhejiang Medical and Health Science and Technology Project

• National Natural Science Foundation of China

Gastrointestinal (GI) cancers such as colorectal cancer (CRC), gastric cancer (GC), esophageal cancer (EG), pancreatic duct adenocarcinoma (PDAC) or hepatocellular cancer (HCC) belong to the most commonly diagnosed types of cancer and are among the most frequent causes of cancer related death worldwide. Most types of GI cancer develop in a stepwise fashion with the occurrence of various driver mutations during tumor progression. Understanding the precise function of mutations driving GI cancer development has been regarded as a prerequisite for an improved clinical management of GI malignancies. During recent years, CRISPR/Cas9 has developed into a powerful tool for genome editing in cancer research by knocking in and knocking out even multiple genes at the same time. Within this review, we discuss recent applications for CRISPR/Cas9-based genome editing in GI cancer research including CRC, GC, EG, PDAC and HCC. These applications include functional studies of candidate genes in cancer cell lines or organoids in vitro as well as in murine cancer models in vivo, library screening for the identification of previously unknown driver mutations and even gene therapy of GI cancers.

• CRISPR/cas9
• cancer of the biliary tract
• colorectal cancer
• esophageal cancer
• gastrointestinal cancer
• hepatocellular cancer
• pancreatic cancer

• MLLs
• histone H3K4
• inhibitors
• methyltransferase
• resistant cancer

• 2020R1A6A1A03043708 National Research Foundation of Korea
• 2020R1I1A1A01051942 National Research Foundation of Korea
• 2017R1D1A1B03031165 National Research Foundation of Korea

Epigenetic mechanisms are central to understanding the molecular basis underlying tumorigenesis. Aberrations in epigenetic modifiers alter epigenomic landscapes and play a critical role in tumorigenesis. Notably, the histone lysine methyltransferase KMT2D (a COMPASS/ Set1 family member; also known as MLL4, ALR, and MLL2) is among the most frequently mutated genes in many different types of cancer. Recent studies have demonstrated how KMT2D loss induces abnormal epigenomic reprograming and rewires molecular pathways during tumorigenesis. These findings also have clinical and therapeutic implications for cancer treatment. In this review, we summarize recent advances in understanding the role of KMT2D in regulating tumorigenesis and discuss therapeutic opportunities for the treatment of KMT2D-deficient tumors.

• KMT2D
• MLL4
• cancer
• epigenetics
• histone methyltransferase

• SMARCA
• SNUC
• SWI/SNF

To understand the effect of metformin on epigenetic regulation, we analyzed histone H3 methylation, DNA methylation, and chromatin accessibility in lung cancer cells. Metformin showed little effect on DNA methylation or chromatin accessibility but significantly reduced H3K4me3 levels at the promoters of positive cell cycle regulatory genes. Metformin downregulated H3K4 methyltransferase MLL2 expression and knockdown of MLL2 resulted in suppression of H3K4me3 expression and lung cancer cell proliferation. We further evaluated the clinicopathological significance of MLL2 in tumor and matched normal tissues from 42 non-small cell lung cancer patients. MLL2 overexpression was significantly associated with poor recurrence-free survival in lung adenocarcinoma. Our study facilitates the understanding of the effect of metformin on the regulation of histone H3K4me3 at promoter regions of cell cycle regulatory genes in lung cancer cells, and MLL2 may be a potential therapeutic target for lung cancer therapy.

This study aimed at understanding the effect of metformin on histone H3 methylation, DNA methylation, and chromatin accessibility in lung cancer cells. Metformin significantly reduced H3K4me3 level at the promoters of positive cell cycle regulatory genes such as CCNB2, CDK1, CDK6, and E2F8. Eighty-eight genes involved in cell cycle showed reduced H3K4me3 levels in response to metformin, and 27% of them showed mRNA downregulation. Metformin suppressed the expression of H3K4 methyltransferases MLL1, MLL2, and WDR82. The siRNA-mediated knockdown of MLL2 significantly downregulated global H3K4me3 level and inhibited lung cancer cell proliferation. MLL2 overexpression was found in 14 (33%) of 42 NSCLC patients, and a Cox proportional hazards analysis showed that recurrence-free survival of lung adenocarcinoma patients with MLL2 overexpression was approximately 1.32 (95% CI = 1.08–4.72; p = 0.02) times poorer than in those without it. Metformin showed little effect on DNA methylation and chromatin accessibility at the promoter regions of cell cycle regulatory genes. The present study suggests that metformin reduces H3K4me3 levels at the promoters of positive cell cycle regulatory genes through MLL2 downregulation in lung cancer cells. Additionally, MLL2 may be a potential therapeutic target for reducing the recurrence of lung adenocarcinoma.

• H3K4 methylation
• MLL2
• cell cycle
• lung cancer
• metformin

• EMT
• Kazakh's ESCC
• MALAT1
• TGF-β1

• Epithelial-mesenchymal transition
• LncRNA GAS5
• PTEN
• microRNA 106b

• CRISPR
• Cancer
• Epithelial–mesenchymal transition
• Gene editing

• CRISPR-Cas Systems/genetics
• Embryonic Development/genetics
• Epithelial-Mesenchymal Transition/genetics
• Gene Editing/methods
• Humans
• Organogenesis/genetics
• Signal Transduction/genetics

Paired box 6 (PAX6) is a transcription factor that has oncogenic features. In breast cancer, PAX6 facilitates tumor progression; however, the underlying mechanism is largely unknown. The majority of breast cancer-related mortalities are associated with metastasis of cancer cells. Therefore, the present study aimed to investigate the role of PAX6 in breast tumor metastasis. PAX6 was stably overexpressed in breast cancer cells to perform tumor migration and metastasis assays in vitro and in vivo. In addition, the expression of PAX6 and transforming growth factor β (TGF-β)-SMAD signaling associated proteins on human breast cancer tissue array, as well as key factors involved in epithelial-mesenchymal transition (EMT) were assayed to explore the mechanism underlying metastasis of breast cancer cells. The expression levels of PAX6 were demonstrated to be increased in human breast cancer tissues and associated with poor clinical outcomes. Overexpression of PAX6 markedly promoted metastasis. Further investigation revealed that PAX6 overexpression increased TGF-β-SMAD signaling pathway and induced EMT. These results suggested that highly expressed PAX6 led to EMT through TGF-β-SMAD signaling pathway, thereby promoting cell metastasis and ultimately affecting survival in patients with breast cancer. Taken together, findings indicated that PAX6 may serve as a therapeutic target for the clinical treatment of breast cancer and the underlying mechanism could be used to overcome metastasis of cancer cells.

Acute lymphoblastic leukemia (ALL), the most common childhood malignancy, is characterized by recurring structural chromosomal alterations and genetic alterations, whose detection is critical in diagnosis, risk stratification and prognostication. However, the genetic mechanisms that give rise to ALL remain poorly understood.

Using next-generation sequencing (NGS) in matched germline and tumor samples from 140 pediatric Chinese patients with ALL, we landscaped the gene mutations and estimated the mutation frequencies in this disease.

Our results showed that the top driver oncogenes having a mutation prevalence over 5% in childhood ALL included KRAS (8.76%), NRAS (6.4%), FLT3 (5.7%) and KMT2D (5.0%). While the most frequently mutated genes were KRAS, NRAS and FLT3 in B cell ALL (B-ALL), the most common mutations were enriched in NOTCH1 (23.1%), FBXW7 (23.1%) and PHF6 (11.5%) in T cell ALL (T-ALL). These mutant genes are involved in key molecular processes, including the Ras pathway, the Notch pathway, epigenetic modification, and cell-cycle regulation. Strikingly, more than 50% of mutations occurred in the high-hyperdiploid (HeH) ALL existed in Ras pathway, especially FLT3 (20%). We also found that the epigenetic regulator gene KMT2D, which is frequently mutated in ALL, may be involved in driving leukemia transformation, as evidenced by an in vitro functional assay.

Overall, this study provides further insights into the genetic basis of ALL and shows that Ras mutations are predominant in childhood ALL, especially in the high-hyperdiploid subtype in our research.

• Acute lymphoblastic leukemia
• Genomics
• KMT2D
• Molecular pathogenesis
• Pediatrics

Background: Esophageal squamous cell carcinoma (ESCC) is a common cancer with poor prognosis. The molecular pathogenesis underlying ESCC remains to be explored. Leucine-rich ɑ-2-glycoprotein 1 (LRG1) has been implicated in the pathogenesis of various cancer types, however its role in ESCC is unknown.

Materials and Methods: Data from the public database was analyzed to address the expression of LRG1 in ESCC. Gain-of-function studies were performed in select ESCC cell lines by over-expression or addition of recombinant LRG1, while loss-of-function studies achieved by small interfering RNA mediated knockdown. Wound healing and transwell assays were conducted to investigate ESCC cell migration and invasion upon manipulating LRG1 levels. Western blot and Immunofluorescence staining were used to examine the changes in epithelial to mesenchymal transition (EMT) and TGFβ signaling pathway.

Results: LRG1 mRNA levels were found to be significantly down-regulated in patients with ESCC as well as in several ESCC cell lines. Silencing of LRG1 promoted, while overexpression of LRG1 inhibited ESCC cell migration and invasion. In line with this, Silencing of LRG1 enhanced, while overexpression of LRG1 reduced TGFβ signaling and EMT of ESCC cells.

Conclusion/Significance: LRG1 suppresses ESCC cell migration and invasion via negative modulation of TGFβ signaling and EMT. Down-regulation of LRG1 in ESCC patients may favor tumor metastasis and disease progression.

• Epithelial to Mesenchymal Transition
• Esophageal Squamous Cell Carcinoma
• Invasion
• LRG1
• Migration
• TGFβ signaling

• CLDN8
• Colon cancer
• Growth
• IL22
• LncRNA LINC00662
• Metastasis
• miR-340-5p

• 3' Untranslated Regions
• Animals
• Cell Line, Tumor
• Cell Movement
• Cell Proliferation
• Claudins/genetics
• Claudins/metabolism
• Colonic Neoplasms/genetics
• Colonic Neoplasms/metabolism
• Colonic Neoplasms/pathology
• Female
• Gene Expression Regulation, Neoplastic
• HCT116 Cells
• Humans
• Interleukins/genetics
• Interleukins/metabolism
• MAP Kinase Signaling System
• Male
• Mice
• Mice, Nude
• MicroRNAs/genetics
• Neoplasm Metastasis
• Neoplasm Transplantation
• Prognosis
• RNA, Long Noncoding/genetics
• Up-Regulation
• Interleukin-22

• mixed-lineage leukemia 2
• oral squamous cell carcinoma
• prognosis
• tumor-infiltrating lymphocytes

Novel therapeutic strategies are urgently needed for patients with a delayed diagnosis of pancreatic ductal adenocarcinoma (PDAC) in order to improve their chances of survival. Recent studies have shown potent anti-neoplastic effects of curcumin and its analogues. In addition, the role of histone methyltransferases on cancer therapeutics has also been elucidated. However, the relationship between these two factors in the treatment of pancreatic cancer remains unknown. Our working hypothesis was that L48H37, a novel curcumin analog, has better efficacy in pancreatic cancer cell growth inhibition in the absence of histone-lysine N-methyltransferase 2D (KMT2D).

To determine the anti-cancer effects of L48H37 in PDAC, and the role of KMT2D on its therapeutic efficacy.

The viability and proliferation of primary (PANC-1 and MIA PaCa-2) and metastatic (SW1990 and ASPC-1) PDAC cell lines treated with L48H37 was determined by CCK8 and colony formation assay. Apoptosis, mitochondrial membrane potential (MMP), reactive oxygen species (ROS) levels, and cell cycle profile were determined by staining the cells with Annexin-V/7-AAD, JC-1, DCFH-DA, and PI respectively, as well as flow cytometric acquisition. In vitro migration was assessed by the wound healing assay. The protein and mRNA levels of relevant factors were analyzed using Western blotting, immunofluorescence and real time-quantitative PCR. The in situ expression of KMT2D in both human PDAC and paired adjacent normal tissues was determined by immunohistochemistry. In vivo tumor xenografts were established by injecting nude mice with PDAC cells. Bioinformatics analyses were also conducted using gene expression databases and TCGA.

L48H37 inhibited the proliferation and induced apoptosis in SW1990 and ASPC-1 cells in a dose- and time-dependent manner, while also reducing MMP, increasing ROS levels, arresting cell cycle at the G2/M stages and activating the endoplasmic reticulum (ER) stress-associated protein kinase RNA-like endoplasmic reticulum kinase/eukaryotic initiation factor 2α/activating transcription factor 4 (ATF4)/CHOP signaling pathway. Knocking down ATF4 significantly upregulated KMT2D in PDAC cells, and also decreased L48H37-induced apoptosis. Furthermore, silencing KMT2D in L48H37-treated cells significantly augmented apoptosis and the ER stress pathway, indicating that KMT2D depletion is essential for the anti-neoplastic effects of L48H37. Administering L48H37 to mice bearing tumors derived from control or KMT2D-knockdown PDAC cells significantly decreased the tumor burden. We also identified several differentially expressed genes in PDAC cell lines expressing very low levels of KMT2D that were functionally categorized into the extrinsic apoptotic signaling pathway. The KMT2D high- and low-expressing PDAC patients from the TCGA database showed similar survival rates,but higher KMT2D expression was associated with poor tumor grade in clinical and pathological analyses.

L48H37 exerts a potent anti-cancer effect in PDAC, which is augmented by KMT2D deficiency.

• Pancreatic neoplasms
• Curcumin analog
• Histone methyltransferase 2D
• Therapeutic effects
• Bioinformatics

The purpose of this study was mainly to explore the role and mechanism of microRNA-18a-5p (miR-18a-5p) in oral squamous cell carcinoma (OSCC). The expression of miR-18a-5p in OSCC cells and normal cells was firstly detected using reverse transcription-quantitative polymerase chain reaction (RT-qPCR). The cell viability, apoptosis, migration and invasion abilities of OSCC cells were determined by MTT, cell apoptosis, wound healing and Transwell assays respectively. Additionally, bioinformatics software analysis and luciferase reporter assays were performed to predict and confirm the candidate target of miR-18a-5p. Western blot analysis was used to assess protein expression. It was revealed that the expression of miR-18a-5p in OSCC cells was higher than that in normal cells. In vitro studies revealed that the cell viability, migration and invasion abilities of OSCC cells were promoted and cell apoptosis was inhibited by miR-18a-5p overexpression. In addition, Smad2 was identified as a target of miR-18a-5p. It was also revealed that miR-18a-5p overexpression significantly inhibited the expression of Smad2, Smad4 and E-cadherin, and the levels of Smad7, collagen I, transforming growth factor-β (TGF-β), α-smooth muscle actin (α-SMA), vimentin were enhanced. While miR-18a-5p downregulation presented the opposite effects. In conclusion, the results indicated that miR-18a-5p can regulate the biological process of OSCC by targeting Smad2 and miR-18a-5p/Smad2 may be potential therapeutic targets for OSCC.

• Chromatin
• Co-occurrence
• Epigenetic
• Lysine methyltransferase

• Bladder cancer
• H3K4me1
• KMT2D
• Metastasis
• Tumor suppressor