Polysaccharide isolated from persimmon leaves (Diospyros kaki Thunb.) suppresses TGF-β1-induced epithelial-to-mesenchymal transition in A549 cells

2020 ◽  
Vol 164 ◽  
pp. 3835-3845
Author(s):  
Won-Chul Lim ◽  
Jae Woong Choi ◽  
Nho-Eul Song ◽  
Chang-Won Cho ◽  
Young Kyoung Rhee ◽  
...  
Keyword(s):  
Epithelial To Mesenchymal Transition ◽  
A549 Cells ◽  
Diospyros Kaki ◽  
Mesenchymal Transition ◽  
Tgf Β1

miR-30a-5p Inhibits Epithelial-to-Mesenchymal Transition by Targeting CDK6 in Nasal Polyps

2020 ◽  
pp. 194589242093981 ◽  
Author(s):  
Ting Zhang ◽  
Yong Zhou ◽  
Bo You ◽  
Yiwen You ◽  
Yongbing Yan ◽  
...  
Keyword(s):  
Nasal Polyps ◽  
Epithelial To Mesenchymal Transition ◽  
Transforming Growth Factor ◽  
A549 Cells ◽  
Expression Level ◽  
Luciferase Reporter ◽  
Aberrant Expression ◽  
Mesenchymal Transition ◽  
Emt Markers ◽  
Tgf Β1

Background Epithelial-to-Mesenchymal Transition (EMT) is considered as a crucial event in disease development and dysregulation of microRNAs (miRNAs) is involved in the regulation of EMT in various human diseases. Emerging evidences congregated over the years have demonstrated that miR-30a-5p was decreased in diseases and its overexpression inhibited the process of diseases via attenuating EMT. Although aberrant expression of miRNAs and occurrence of EMT were previously reported in Nasal Polyps (NPs), the role of miR-30a-5p in EMT of NPs is still remains unclear. Objective The purpose of our present study was to explore the expression and potential function of miR-30a-5p in EMT of NPs. Methods The expression of miR-30a-5p and mRNA expression level were detected by quantitative real-time PCR (qRT-PCR) in transforming growth factor β1 (TGF-β1) - induced EMT model and NPs patients. Western Blot (WB) and immunohistochemistry (IHC) were performed to evaluate the protein expression level of EMT markers. The cells mobility was assessed by Wound-Healing assay. Luciferase reporter assay was utilized to verify the relationship between Cyclin-dependent kinase 6 (CDK6) and miR-30a-5p. Results Firstly, we observed that miR-30a-5p was down-regulated notably, accompanying with the alteration of EMT markers expression in NPs tissues and EMT model induced by TGF-β1 in primary Human Nasal Epithelial Cells (pHNECs) and A549 cells in vitro. Moreover, the functional assays demonstrated that overexpression of miR-30a-5p significantly inhibited EMT and cells mobility. Subsequently, CDK6 was validated as a direct target of miR-30a-5p. Finally, we performed the rescue experiments indicating that overexpression of CDK6 eliminated the suppressive effects of miR-30a-5p in TGF-β1-induced EMT in pHNECs and A549 cells. Conclusion Taken together, our results suggested that EMT was involved in NPs, and overexpression of miR-30a-5p could attenuate EMT via repressing the expression of the CDK6 in pHNECs and A549 cells.

TGF-β1 induces epithelial-to-mesenchymal transition via inhibiting mitochondrial functions in A549 cells

2018 ◽  
Vol 52 (11-12) ◽  
pp. 1432-1444 ◽  
Author(s):  
Jiaxin Zhang ◽  
Wei Zhang ◽  
Tao Zhang ◽  
Qingbiao Zhou ◽  
Jiangzheng Liu ◽  
...  
Keyword(s):  
Epithelial To Mesenchymal Transition ◽  
A549 Cells ◽  
Mesenchymal Transition ◽  
Mitochondrial Functions ◽  
Tgf Β1

scholarly journals The Epithelial-to-Mesenchymal Transition-Like Process Induced by TGF-β1 Enhances Rubella Virus Binding and Infection in A549 Cells via the Smad Pathway

2021 ◽  
Vol 9 (3) ◽  
pp. 662
Author(s):  
Ngan Thi Kim Pham ◽  
Quang Duy Trinh ◽  
Kazuhide Takada ◽  
Chika Takano ◽  
Mari Sasano ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Rubella Virus ◽  
Epithelial To Mesenchymal Transition ◽  
A549 Cells ◽  
Strong Increase ◽  
Flow Cytometry Analysis ◽  
Virus Binding ◽  
Smad Pathway ◽  
Mesenchymal Transition ◽  
Tgf Β1

Virus–host cell interactions in rubella virus (RuV) are of great interest in current research in the field, as their mechanism is not yet well understood. By hypothesizing that the epithelial-to-mesenchymal transition (EMT) may play a role in RuV infection, this study aimed to investigate the influence of TGF-β1-induced EMT of human lung epithelial A549 cells on the infectivity of RuV. A549 cells were cultured and treated with TGF-β1 for 1 to 2 days prior to virus infection (with a clinical strain). Viral infectivity was determined by flow cytometry analysis of cells harvested at 24 and 48 h post-infection (hpi) and by titration of supernatants collected at 48 hpi. The results showed that the percentages of the TGF-β1-treated A549 cells that were positive for RuV were at least twofold higher than those of the control, and the viral progeny titers in the supernatants collected at 48 hpi were significantly higher in the treatment group than in the control group. In addition, the virus binding assay showed a strong increase (more than threefold) in the percentages of RuV-positive cells, as determined by flow cytometry analysis and further confirmed by real-time PCR. Such an enhancement effect on RuV infectivity was abolished using LY364947 or SB431542, inhibitors of the TGF-β/Smad signaling pathway. The findings suggest that the TGF-β1-induced EMT-like process enhances RuV binding and infection in A549 cells via the Smad pathway. Further studies are necessary to identify possible proteins that facilitate viral binding and entry into treated cells.

scholarly journals Overexpression of GRB2 Enhances Epithelial to Mesenchymal Transition of A549 Cells by Upregulating SNAIL Expression

Cells ◽  
2018 ◽  
Vol 7 (8) ◽  
pp. 97 ◽  
Author(s):  
Payal Mitra ◽  
Pazhanichamy Kalailingam ◽  
Hui Tan ◽  
Thirumaran Thanabalu
Keyword(s):  
Epithelial To Mesenchymal Transition ◽  
A549 Cells ◽  
Adaptor Protein ◽  
Sh3 Domain ◽  
Sh2 Domain ◽  
Migration And Invasion ◽  
Mesenchymal Transition ◽  
Β Receptor ◽  
Tgf Β1 ◽  
E Cadherin

GRB2 is an adaptor protein which interacts with phosphorylated TGF-β receptor and is critical for mammary tumour growth. We found that TGF-β1-induced EMT increased GRB2 expression in A549 cells (non-small cell lung cancer). Overexpression of GRB2 (A549GRB2) enhanced cell invasion while knocking down GRB2 (A549GRB2KD) reduced cell migration and invasion, probably due to increased vinculin and reduced Paxillin patches in A549GRB2KD cell. TGF-β1-induced EMT was more pronounced in A549GRB2 cells and attenuated in A549GRB2KD cells. This could be due to the reduced expression of E-cadherin in A549GRB2 and increased expression of E-cadherin in A549GRB2KD cells, even before TGF-β1 stimulation. Expression of SNAIL was elevated in A549GRB2 cells and was further enhanced by TGF-β1 stimulation, suggesting that GRB2 down-regulates E-cadherin by enhancing the expression of SNAIL. The N-SH3 domain of GRB2 was critical for suppressing E-cadherin expression, while the C-SH3 domain of GRB2 mediating interaction with proteins such as N-WASP was critical for promoting invasion, and the SH2 domain was critical for suppressing E-cadherin expression and invasion. Thus, our data suggests that GRB2 enhances EMT by suppressing E-cadherin expression and promoting invasion probably through N-WASP to promote metastasis.

scholarly journals Ovalitenone Inhibits the Migration of Lung Cancer Cells via the Suppression of AKT/mTOR and Epithelial-to-Mesenchymal Transition

Molecules ◽  
2021 ◽  
Vol 26 (3) ◽  
pp. 638
Author(s):  
Kittipong Sanookpan ◽  
Nongyao Nonpanya ◽  
Boonchoo Sritularak ◽  
Pithi Chanvorachote
Keyword(s):  
Lung Cancer ◽  
Cell Migration ◽  
Cancer Cells ◽  
Colony Formation ◽  
Cancer Metastasis ◽  
Epithelial To Mesenchymal Transition ◽  
A549 Cells ◽  
Lung Cancer Cells ◽  
Migration And Invasion ◽  
Mesenchymal Transition

Cancer metastasis is the major cause of about 90% of cancer deaths. As epithelial-to-mesenchymal transition (EMT) is known for potentiating metastasis, this study aimed to elucidate the effect of ovalitenone on the suppression of EMT and metastasis-related behaviors, including cell movement and growth under detached conditions, and cancer stem cells (CSCs), of lung cancer cells. Methods: Cell viability and cell proliferation were determined by 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazo-liumbromide (MTT) and colony formation assays. Cell migration and invasion were analyzed using a wound-healing assay and Boyden chamber assay, respectively. Anchorage-independent cell growth was determined. Cell protrusions (filopodia) were detected by phalloidin-rhodamine staining. Cancer stem cell phenotypes were assessed by spheroid formation. The proteins involved in cell migration and EMT were evaluated by Western blot analysis and immunofluorescence staining. Results: Ovalitenone was used at concentrations of 0–200 μM. While it caused no cytotoxic effects on lung cancer H460 and A549 cells, ovalitenone significantly suppressed anchorage-independent growth, CSC-like phenotypes, colony formation, and the ability of the cancer to migrate and invade cells. The anti-migration activity was confirmed by the reduction of filopodia in the cells treated with ovalitenone. Interestingly, we found that ovalitenone could significantly decrease the levels of N-cadherin, snail, and slug, while it increased E-cadherin, indicating EMT suppression. Additionally, the regulatory signaling of focal adhesion kinase (FAK), ATP-dependent tyrosine kinase (AKT), the mammalian target of rapamycin (mTOR), and cell division cycle 42 (Cdc42) was suppressed by ovalitenone. Conclusions: The results suggest that ovalitenone suppresses EMT via suppression of the AKT/mTOR signaling pathway. In addition, ovalitenone exhibited potential for the suppression of CSC phenotypes. These data reveal the anti-metastasis potential of the compound and support the development of ovalitenone treatment for lung cancer therapy.

scholarly journals Serum containing drugs of Gua Lou Xie Bai decoction (GLXB-D) can inhibit TGF-β1-Induced Epithelial to Mesenchymal Transition (EMT) in A549 Cells

2018 ◽  
Vol 16 (1) ◽  
pp. 407-414
Author(s):  
Rui-qin Li ◽  
Bai-yan Wang ◽  
Yu-wen Ding ◽  
Rui Zhang ◽  
Jun-xia Zhang ◽  
...  
Keyword(s):  
Pulmonary Fibrosis ◽  
Transforming Growth Factor ◽  
A549 Cells ◽  
Alveolar Epithelial Cells ◽  
Collagen I ◽  
Potential Candidate ◽  
Mesenchymal Transition ◽  
Thiazolyl Blue Tetrazolium Bromide ◽  
Tgf Β1 ◽  
E Cadherin

AbstractThe present study explores the mechanism of resistance to pulmonary fibrosis by observing the possible effects of serum containing drugs prepared from Gua Lou Xie Bai decoction (GLXB-D) on transforming growth factor beta 1 (TGF-β1) induced Epithelial-mesenchymal transition (EMT) of A549 human alveolar epithelial cells. The inhibition rate was observed with the help of thiazolyl blue tetrazolium bromide (MTT) in 24 h and 48 h treated cells. Inverted microscope and transmission electron microscope (TEM) were used to study the changes in the morphology and ultrastructure of the cells. The expressions of E-cadherin and Vimentin were comparatively analyzed by Western blotting, while the expressions of Collagen I and III were analyzed by ELISA. The data obtained indicated that the expression of epithelial marker E-cadherin was decreased, while the expressions of EMT markers such as Vimentin and Collagen I and III were increased in 24 h after TGF-β1 induction. However, the serum containing drugs of GLXB-D were found to inhibit the TGF-β1 induced proliferation of cells, increase the expression of E-cadherin and decrease the expression of Vimentin, collagen I and III. In conclusion, the serum containing drugs of GLXB-D effectively reduced pulmonary fibrosis, mainly via the reversal of EMT induction by TGF-β1. Thus, it can be considered as a potential candidate for the development of better treatment methods for pulmonary fibrosis.

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