CBX8 Promotes Epithelial-mesenchymal Transition, Migration, and Invasion of Lung Cancer through Wnt/β-catenin Signaling Pathway.

Lung cancer (LC) is primarily responsible for cancer-related deaths worldwide. Epithelial-mesenchymal transition (EMT) is a process in which epithelial cells acquire mesenchymal features and is associated with the development of tumors. CBX8, a member of the PcG protein family, plays a critical role in various cancers, containing LC. However, specific regulatory mechanisms of CBX8 in LC progression are not fully understood. This study aimed to investigate the regulatory role of CBX8 in LC progression. Bioinformatics was used to analyze the relationship between CBX8 level and tumor and the enrichment pathway of CBX8 enrichment. qRT-PCR was used to detect the differential expression of CBX8 in LC cells and normal lung epithelial cells. The effects of knockdown or overexpression of CBX8 on the proliferation, migration and invasion of LC cells were evaluated by CCK- -8 assay and Transwell assay, and the levels of proteins associated with the EMT pathway and Wnt/ β-catenin signaling pathway were detected by western blot. Bioinformatics analysis revealed that CBX8 was highly expressed in LC and enriched on the Wnt/β-catenin signaling pathway. The expression level of CBX8 was significantly elevated in LC cells. Knockdown of CBX8 significantly inhibited cell proliferation, migration and invasion, and decreased the expression levels of EMT-related proteins and Wnt/β-catenin pathway-related proteins. Conversely, overexpression of CBX8 promoted cell proliferation, migration and invasion, and increased the expression levels of EMT-related proteins and Wnt/β-catenin pathway-related proteins. The Wnt inhibitor IWP-4 alleviated the effects produced by overexpression of CBX8. Collectively, these data demonstrated that CBX8 induced EMT through Wnt/β-- catenin signaling, driving migration and invasion of LC cells.

Cai X ，Lv Y ，Pan J ，Cao Z ，Zhang J ，Li Y ，Zheng H ... -  《-》

ING5 inhibits lung cancer invasion and epithelial-mesenchymal transition by inhibiting the WNT/β-catenin pathway.

ING5 is the last member of the Inhibitor of Growth (ING) candidate tumor suppressor family that has been implicated in multiple cellular functions, including cell cycle regulation, apoptosis, and chromatin remodeling. Our previous study showed that ING5 overexpression inhibits lung cancer aggressiveness and epithelial-mesenchymal transition (EMT), with unknown mechanisms. Western blotting was used to detect total and phosphorylated levels of β-catenin and EMT-related proteins. Immunofluorescent staining was used to observe E-cadherin expression. Proliferation and colony formation, wound healing, and Transwell migration and invasion assays were performed to study the proliferative and invasive abilities of cancer cells. ING5 overexpression promotes phosphorylation of β-catenin at Ser33/37, leading to a decreased β-catenin protein level. Small hairpin RNA-mediated ING5 knockdown significantly increased the β-catenin level and inhibited phosphorylation of β-catenin S33/37. Treatment with the WNT/β-catenin inhibitor XAV939 inhibited ING5-knockdown promoted proliferation, colony formation, migration, and invasion of lung cancer A549 cells, with increased phosphorylation of β-catenin S33/37 and a decreased β-catenin level. XAV939 also impaired ING5-knockdown-induced EMT, as indicated by upregulated expression of the EMT marker E-cadherin, an epithelial marker; and decreased expression of N-cadherin, a mesenchymal marker, and EMT-related transcription factors, including Snail, Slug, Twist, and Smad3. Furthermore, XAV939 could inhibit the activation of both IL-6/STAT3 and PI3K/Akt signaling pathways. ING5 inhibits lung cancer invasion and EMT by inhibiting the WNT/β-catenin pathway.

Liu XL ，Meng J ，Zhang XT ，Liang XH ，Zhang F ，Zhao GR ，Zhang T ... -  《-》

TNNT1 accelerates migration, invasion and EMT progression in lung cancer cells.

Clinically, most patients with lung cancer (LC) die from tumor spread and metastasis. Specific metastasis-related molecules can provide reference for clinical prediction of efficacy, evaluation of prognosis, and search for the best treatment plan. Troponin T1 (TNNT1) is highly expressed in various cancer tissues, which affects malignant behavior of tumor cells and is related to patients' survival and prognosis. However, the role and molecular mechanism of TNNT1 in LC invasion and metastasis have not yet been investigated. Gene expression profiling interactive analysis (GEPIA) online analysis was used to analyze TNNT1 expression in LC tissues. Quantitative real-time-polymerase chain reaction (qRT-PCR) or western blot were performed to measure TNNT1 or epithelial-to-mesenchymal transition (EMT)-related and Wnt/β-catenin pathway-related protein expression in LC cells. After TNNT1 knockdown, cell scratch healing and transwell assays were introduced to assess cell migration and invasion, respectively. TNNT1 expression in LC tissues and cells was increased. TNNT1 knockdown notably impaired LC cell migration, invasion and EMT. TNNT1 knockdown inhibited Wnt/β-catenin pathway of LC cells. Lithium chloride (LiCl) addition partially restored the inhibition of TNNT1 knockdown on migration, invasion, EMT and Wnt/β-catenin of LC cells. TNNT1 knockdown attenuated LC migration, invasion and EMT, possibly through Wnt/β-catenin signaling.

Ge X ，Du G ，Zhou Q ，Yan B ，Yue G ... -  《-》

Forkhead Box S1 mediates epithelial-mesenchymal transition through the Wnt/β-catenin signaling pathway to regulate colorectal cancer progression.

Recent studies have shown that the fox family plays a vital role in tumorigenesis and progression. Forkhead Box S1 (FOXS1), as a newly identified subfamily of the FOX family, is overexpressed in certain types of malignant tumors and closely associated with patient's prognosis. However, the role and mechanism of the FOXS1 in colorectal cancer (CRC) remain unclear. FOXS1 level in CRC tissues and cell lines was analyzed by western blot and quantitative real-time polymerase chain reaction (qRT-PCR). Immunohistochemistry (IHC) was used to detect the relationship between FOXS1 expression and clinicopathological features in 136 patients in our unit. The expression of FOXS1 was knocked down in CRC cells using small interfering RNA (siRNA) technology. Cell proliferation was assessed by CCK8 assay, colony formation, and 5-Ethynyl-20-deoxyuridine (EdU) incorporation assay. Flow cytometry detected apoptosis and wound healing, and Transwell assays determined cell migration and invasion. Western blotting was used to detect the levels of proteins associated with the Wnt/β-catenin signaling pathway. Then, we used short hairpin RNA (shRNA) to knock down FOXS1 to see the effect of FOXS1 on the proliferation, migration, invasion, and metastasis of CRC cells in vivo. Finally, we investigated the impact of Wnt activator LiCl on the proliferation, migration, invasion, and metastasis of CRC cells after FOXS1 knockdown. Compared to those in normal groups, FOXS1 overexpressed in CRC tissues and CRC cells (P < 0.05). Upregulation of FOXS1 association with poor prognosis of CRC patients. si-FOXS1 induced apoptosis and inhibited proliferation, migration, invasion, the epithelial-mesenchymal transition (EMT), and the Wnt/β-catenin signaling pathway in vitro; sh-FOXS1 inhibited the volume and weight of subcutaneous xenografts and the number of lung metastases in vivo. LiCl, an activator of Wnt signaling, partially reversed the effect of FOXS1 overexpression on CRC cells. FOXS1 could function as an oncogene and promote CRC cell proliferation, migration, invasion and metastasis through the Wnt/βcatenin signaling pathway, FOXS1 may be a potential target for CRC treatment.

Zhang L ，Ren CF ，Yang Z ，Gong LB ，Wang C ，Feng M ，Guan WX ... -  《Journal of Translational Medicine》

Golgi Phosphoprotein 3 Promotes Cervical Cancer Progression via Wnt/β-catenin Mediated Epithelial-Mesenchymal Transition.

Cervical cancer (CC) is one of the fatal malignancies affecting the life expectancy of women worldwide. Golgi Phosphoprotein 3 (GOLPH3) has been shown to play a key role in the development of various tumors. However, the role of GOLPH3 in the development of CC is unclear. GOLPH3 levels were measured using quantitative real-time polymerase chain reaction (qRT-PCR) and western blot assays. Cell Counting Kit-8 (CCK-8) and colony formation assays were used to detect cell proliferation. Xenograft tumor models were used to explore the effects of GOLPH3 on tumor growth of mice, and immunohistochemistry assay was performed to determine the expression of GOLPH3 and Ki-67. Transwell assay was performed to evaluate cell migration and invasion. Western blot assay was used to analyze the signaling molecules-related proteins regulated by GOLPH3. GOLPH3 was upregulated in human CC tissues from the GEO database (GSE39001 and GSE63514), and further demonstrated that GOLPH3 level was elevated in human CC cells. GOLPH3 enhanced CC cell proliferation, and knockdown of GOLPH3 suppressed tumor growth and decreased Ki-67 level in xenograft mice. In addition, GOLPH3 aggravated the migration and invasion of CC cells. The data indicated that Wnt/β-catenin signaling might be one of the key targets of GOLPH3. Blockage of the Wnt/β-catenin pathway by XAV-939 significantly affected the effects of GOLPH3 on cell proliferation and epithelial-mesenchymal transition (EMT) related molecules, whereas LiCl (a Wnt/β-catenin signal activator) reversed these above effects. GOLPH3 promotes cell proliferation, migration and invasion in CC, possibly by regulating the Wnt/β-catenin signaling pathway, which may provide a new idea for the development of CC therapeutic targets.

Yang H ，Ma X ，Song B ，Wang D ，Chai Z ... -  《-》