Anti-tumor Drug THZ1 Suppresses TGFβ2-mediated EMT in Lens Epithelial Cells via Notch and TGFβ/Smad Signaling Pathway

Abstract

Selective covalent CDK7 inhibitor THZ1 is a promising potential anti-tumor drug in many kinds of cancers. Epithelial-mesenchymal Transition (EMT) is highly related to cancer initiation, development, invasion and metastasis and other pathogenesis processes. We treated cancer cell line Hela229 and three retinoblastoma cell lines so-RB50, WERI-Rb-1, Y79 with gradient concentration of THZ1, and found that THZ1 could inhibit cell viability and EMT, suggesting that THZ1 may be a promising drug for human cervical cancer and retinoblastoma treatment. Our results verified the role of THZ1 in EMT for the first time, however, the mechanism needs further study. Here we report that THZ1 suppresses the TGFβ2 induced EMT in human SRA01/04 lens epithelial cells (LECs), rabbit primary lens epithelial cells, and whole rat lens culture semi-in vivo model. RNA-sequencing and KEGG analysis revealed that the THZ1 inhibits EMT by down-regulating phosphorylate Smad2 and Notch signaling pathway. On the other hand, we found that THZ1 could strongly inhibit LECs proliferation through G2/M phase arrest as well as attenuating of MAPK, PI3K/AKT signaling pathway. Our results uncovered the function and underlying mechanism of THZ1 in regulation of EMT, which provides a new perspective of the anti-tumor effect by THZ1 and may offer a novel treatment for PCO.

Cyclin‐dependent kinase 7 inhibitor THZ1 in cancer therapy

“…2018 46 NSCLCCell proliferation assay; wound-healing assay; qRT-PCR analysis; immunoblot analysis; cell-cycle analysis; apoptosis analysisTHZ1 induced cell cycle arrest; blocked the glycolysis pathway; promoted ubiquitination and degradation of NUDT21 Ning et al. 2019 67 Cervical cancer and retinoblastomaCell-cycle analysis; apoptosis analysis; RT-PCR analysis; Immunoblot analysis; RNA sequencing; In vitro & in vivo assayTHZ1 prevents TGFβ2-induced EMT in human LECs;THZ1 inactivates the MAPK, ERK1/2 and PI3K/AKT signaling pathways; THZ1 caused cell-cycle arrestLu et al. 2019 68 PDACCell viability assay; apoptosis analysis; cell-cycle analysis; immunoblot analysis; immunohistochemical analysis; H&E staining of tissue sections; GO term enrichment analysis; RNA sequencing; in vitro & in vivo assayTHZ1 caused cell-cycle arrest; THZ1 inhibites TNF/NF-κB signaling pathway; THZ1 downregulates Hippo, MAPK signaling pathwaysT-ALL: T-cell acute lymphoblastic leukaemia; GO: gene ontology; ChIP-seq: chromatin immunoprecipitation coupled with high-throughput sequencing; RUNX1 : Runt-related transcription factor 1; NB: neuroblastoma; qRT-PCR: quantitative real-time polymerase chain reaction; PCR: polymerase chain reaction; CHIP-qPCR: chromatin Immunoprecipitation-quantitative PCR; SCLC: small-cell lung cancer; MRI: magnetic resonance imaging; ASCL1 : achaete-scute homolog 1; INSM1 : insulinoma-associated 1; TNBC: triple-negative breast cancer; H & E: hematoxylin-eosin; CRISPR: clusters of regularly interspaced short palindromic repeats; Cas9: CRISPR associated protein 9; CDK7: cyclin-dependent kinase 7; PTCL: peripheral T-cell lymphomas; Q-ChIP: fast chromatin immunoprecipitation assay; RT-PCR: real-time PCR; RNA: ribonucleic acid; STAT: signal transducers and activators of transcription; NSCLC: non-small-cell lung cancer; NUDT21 : nudix-type motif 21; RT-PCR: real-time PCR; TGFβ2: transforming growth factor beta 2; EMT: epithelial-to-mesenchymal transition; LECs: lens epithelial cells; ERK1/2: extracellular signal-regulated kinase1/2; PI3K: phosphatidylinositol 3-kinases; AKT: protein kinase B; PDAC: pancreatic ductal adenocarcinoma; TNF: tumor necrosis factor; NF-κB: nuclear factor-κB; MAPK: mitogen-activated protein kinase.…”

Inhibition of cyclin‐dependent kinase 7 down‐regulates yes‐associated protein expression in mesothelioma cells

“…Several groups reported YAP overexpression promotes epithelial to mesenchymal transition (EMT) in cancer cell lines . Recently, the CDK7 inhibitor THZ1 was suggested to increase the stability of Snail protein in colorectal cancer cell lines, and THZ1 was thought to suppress TGFβ2‐mediated EMT in lens epithelial cells . MPM has three pathological subtypes, epithelial, sarcomatoid and biphasic.…”

Research Progress of Drug Prophylaxis for Lens Capsule Opacification after Cataract Surgery

“…Biochemical reactions of cells are based on a series of different proteins, and each chain of proteins named a signaling pathway. Several signaling pathways, including phosphoinositide 3-kinase (PI3K)/Akt pathway [ 15 ], Rho kinase pathway [ 16 ], Smad pathway [ 17 ], and mitogen-activated protein kinase (MAPK) pathway [ 18 ], have been pointed out to involve the pathogenesis of lens capsule opacification. Kayastha et al [ 15 ] found that both andrographolide and LY294002, a known PI3K inhibitor, could decrease the level of activated Akt kinase by Western blotting and explored that the inhibitory effect of andrographolide on human LECs' proliferation in vitro exhibited dose–response relationship with the IC 50 of 1 μ mol/L via BrdU incorporation assay.…”

“…TGF- β plays a key role in LECs' EMT [ 32 ] and is often used for building LECs' EMT model. Ning et al [ 17 ] discovered that THZ1, a potential antineoplastic agent, could inhibit not only cyclin-dependent kinases of tumor cells, but also the EMT of LECs activated by TGF- β . The result of Western blotting showed that THZ1 lowered the level of active Smad, α -SMA, and fibronectins, which meant the possible mechanism of THZ1 in impeding LECs' EMT is the inhibition of Smad signaling pathway.…”