A novel ETV6-miR-429-CRKL regulatory circuitry contributes to aggressiveness of hepatocellular carcinoma

Guo, Chunmei; Gao, Chao; Zhao, Dongting; Li, Jiahui; Wang, Jinxia; Sun, Xujuan; Liu, Qinlong; Hao, Lihong; Greenaway, Frederick T.; Tian, Ye; Liu, Shuqing; Sun, Ming‐Zhong

doi:10.1186/s13046-020-01559-1

Cited by 16 publications

(45 citation statements)

References 53 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Our previous work also indicated that CRKL upregulation potentially promoted HCC progression by enhancing the in vitro malignant behaviors of HepG2 cells ( Guo et al, 2018 ). The in vitro data on miR-124-3p and CRKL expression as well as invasion and migration activities in HCC cells from current study are consistent with our previous studies ( Guo et al, 2018 , 2020 ).…”

Section: Discussionsupporting

confidence: 92%

“…Twenty three frozen HCC human tissues including their corresponding non-tumor liver tissues (14 male and 9 female patients, 8 patients under ≥ 60 age and 15 < 60 age) were used for comparative miR-124-3p expression analysis. However, the CRKL protein expression analysis from these tissues was already reported in our previous work ( Guo et al, 2018 , 2020 ). In the current work, we quantified miR-124-3p in these tissues and correlated miR-124-3p expression from the current study, with CRKL expression from a previous study.…”

Section: Methodsmentioning

confidence: 81%

“…In the current work, 23 frozen HCC human tissues including their corresponding non-tumor liver tissues were used for miR-124-3p. The CRKL protein expression analysis from these tissues was reported in our previous work ( Guo et al, 2018 , 2020 ). In the current study, we quantified miR-124-3p in these tissues and correlated with tissue expression of CRKL from a previous study.…”

Section: Introductionmentioning

confidence: 94%

See 2 more Smart Citations

miR-124-3p Suppresses the Invasiveness and Metastasis of Hepatocarcinoma Cells via Targeting CRKL

Majid

Wang

Nawaz

et al. 2020

Front. Mol. Biosci.

Self Cite

View full text Add to dashboard Cite

Abnormal expressions of microRNAs are involved in growth and progression of human cancers including hepatocellular carcinoma (HCC). An adaptor protein CRKL plays a pivotal role in HCC growth, whereas miR-124-3p downregulation is associated with clinical stage and the poor survival of patients. However, the relationship between miR-124-3p and CRKL and the molecular mechanisms through which they regulate HCC metastasis remains unclear. In the current work, we explored miR-124-3p and its correlation with CRKL expression in HCC patient tissues. We found that miR-124-3p deficiency is inversely co-related with CRKL overexpression in tumorous tissues of HCC patients, which was also consistent in HCCLM3 and Huh7 HCC cell lines. Target validation data shows that miR-124-3p directly targets CRKL. The overexpression of miR-124-3p reverses the CRKL expression at both mRNA and protein levels and inhibits the cell development, migration, and invasion. Mechanistic investigations showed that CRKL downregulation suppresses the ERK pathway and EMT process, and concomitant decrease in invasion and metastasis of HCC cells. The expressions of key molecules in the ERK pathway such as RAF, MEK, ERK1/2, and pERK1/2 and key promoters of EMT such as N -cadherin and vimentin were downregulated, whereas E -cadherin, a key suppression indicator of EMT, was upregulated. MiR-124-3p-mediated CRKL suppression led to BAX/BCL-2 increase and C-JUN downregulation, which inhibited the cell proliferation and promoted the apoptosis in HCC cells. Collectively, our data illustrates that miR-124-3p acts as an important tumor-suppressive miRNA to suppress HCC carcinogenesis through targeting CRKL. The miR-124-3p-CRKL axial regulated pathway may offer valuable indications for cancer research, diagnosis, and treatment.

show abstract

Section: Discussionsupporting

confidence: 92%

Section: Methodsmentioning

confidence: 81%

Section: Introductionmentioning

confidence: 94%

See 1 more Smart Citation

miR-124-3p Suppresses the Invasiveness and Metastasis of Hepatocarcinoma Cells via Targeting CRKL

Majid

Wang

Nawaz

et al. 2020

Front. Mol. Biosci.

Self Cite

View full text Add to dashboard Cite

show abstract

“…CRKL is a major tyrosine-phosphorylated protein in CML cells, pCRKL plays a special role in CML pathogenesis, and the constitutive phosphorylation of CRKL is unique to CML, which makes it a useful target for therapeutic intervention [42][43][44]. We previous reported that CRKL is associated with proliferation, migration and invasion of hepatocarcinoma and clear cell renal cell carcinoma cells [45][46][47][48][49][50]. However, the exact role of CRKL in CML is unknown.…”

Section: Discussionmentioning

confidence: 99%

CRKL but not CRKII Inhibits Erythropoiesis and Megakaryopoiesis of CML via Inactivating Raf/MEK/ERK/Elk-1 Pathway

Guo

Zhang

Yan

et al. 2020

Preprint

Self Cite

View full text Add to dashboard Cite

Background: As members of the CT10 regulation of kinase (CRK) adaptor protein family, CRK-like (CRKL) and CRKII are involved in cell proliferation, survival, adhesion, migration and differentiation. However, the exact role and underlying mechanism of CRKL and CRKII in leukemic cell differentiation are still unknown. Methods: Quantitative real-time qPCR (qRT-PCR) was used to detect the expression levels of CRKL and CRKII in chronic myeloid leukemia (CML) patients and complete remission (CR) patients; Western blotting (WB) was used to measure the expression levels of CRKL and CRKII during hemin-induced erythroid differentiation of K562 cells; Benzidine staining, isobaric tags for relative and absolute quantitation (iTRAQ) proteomic analysis, cDNA microarray assay, qRT-PCR and WB were used to examine the effects of CRKL and CRKII deregulation on erythroid and megakaryocyte differentiation of K562 cells; PD98059 was used to investigate the underlying mechanism of CRKL in erythropoiesis and megakaryopoiesis. Results: CRKL was found to be overexpressed in chronic myeloid leukemia (CML) patients compared with normal samples, while its expression level was lower in CR patients than in corresponding CML patients. The CRKL expression level was significantly decreased during the erythroid differentiation of K562 cells following hemin treatment. Moreover, CRKL downregulation promoted erythroid and megakaryocyte differentiation of K562 cells accompanied by increased expression level of the erythroid differentiation markers γ-globin, glycophorin (GPA) and the megakaryocyte differentiation markers CD41, CD61. Furthermore, gene microarray and iTRAQ quantitative proteomic analysis showed that CRKL downregulation increased hemoglobin (HB) molecules HBD, HBA1, HBA2, HBZ, HBE1, HBG1 and globin transcription factor 1 (GATA1), high-mobility group protein (HMGB2) expression levels. Mechanistically, CRKL inhibited erythroid and megakaryocyte differentiation of K562 cell via inactivating Raf/MEK/ERK/Elk-1 pathway. Conversely, CRKII was only slightly overexpressed in CML patients and had no effect on erythroid differentiation of K562 cells. Conclusions: Taken together, our results demonstrate that CRKL but not CRKII contributes to development, progression, erythropoiesis and megakaryopoiesis of CML, providing novel insights into effective diagnosis and therapy for CML patients.

show abstract

“…For IHC staining, the experiment was performed using manufacturer's instructions for kits SP-9001 (Zhong Shan-Golden Bridge Biological Technology). IHC assays were scored as previously describe [36].…”

Section: Immunocytochemical (Icc) and Immunohistochemical (Ihc) Stainingmentioning

confidence: 99%

Nrf2 overexpression increases risk of high tumor mutation burden in acute myeloid leukemia through inhibiting MSH2

Liu

Wang

et al. 2020

Preprint

View full text Add to dashboard Cite

Background: Nuclear factor erythroid 2-related factor 2 (Nrf2, also called NFE2L2) has been shown to play a pivotal role in preventing cancer cells from being affected by chemotherapy. Gene mutation is a crucial reason of chemotherapy-resistance in acute myeloid leukemia (AML). However, the relationship between Nrf2 and tumor mutation burden and its mechanism in regulating chemotherapy-resistance remains unclear. Methods: The whole-exome sequencing analysis were used to measure tumor mutation. RNA sequencing, Oncomine, qRT-PCR, Western blotting and immunocytochemistry were employed to detect differences in genes and proteins. The KEGG pathway enrichment analysis and GeneMANIN were performed pathway analysis. Functional assays, such as annexin V/PI, Hoechst33342 staining and DCFH were performed to examine the apoptosis and reactive oxygen species (ROS) of AML cells in vitro. Subcutaneous xenograft model was established to investigate in vivo growth. Results: Nrf2 expression was associated with tumor mutation burden in AML. Patients with Nrf2 overexpression had higher frequency of gene mutation and drug resistance. Nrf2 overexpression protected the AML cells from apoptosis induced by cytarabine in vitro and increased the risk of gene mutant drug resistance in vivo. Furthermore, Nrf2 overexpression inhibited MSH2 protein expression, which caused DNA mismatch repair (MMR) deficiency. Mechanistically, the inhibition of MSH2 by Nrf2 was in a ROS-independent manner. Further studies showed that an increased activation of JNK/c-Jun signaling in Nrf2 overexpression cells, which inhibited the expression of MSH2 protein. Conclusions: Our findings provided evidence that high Nrf2 expression inhibited MSH2 expression, caused MMR deficiency and increased the tumor mutation burden, which can induce gene instability-dependent drug resistance in AML. This study demonstrates the reason why the high Nrf2 expression leads to the increase of gene mutation frequency in AML, and provides a new strategy for clinical practice.

show abstract

A novel ETV6-miR-429-CRKL regulatory circuitry contributes to aggressiveness of hepatocellular carcinoma

Cited by 16 publications

References 53 publications

miR-124-3p Suppresses the Invasiveness and Metastasis of Hepatocarcinoma Cells via Targeting CRKL

miR-124-3p Suppresses the Invasiveness and Metastasis of Hepatocarcinoma Cells via Targeting CRKL

CRKL but not CRKII Inhibits Erythropoiesis and Megakaryopoiesis of CML via Inactivating Raf/MEK/ERK/Elk-1 Pathway

Nrf2 overexpression increases risk of high tumor mutation burden in acute myeloid leukemia through inhibiting MSH2

Contact Info

Product

Resources

About