Ca2+-activated Cl- channel TMEM16A inhibition by cholesterol promotes angiogenesis in endothelial cells
© 2021 The Authors. Published by Elsevier B.V. on behalf of Cairo University..
Introduction: Ca2+-activated Cl- channel TMEM16A is expressed in endothelial cells, and contributes to many diseases such as hypertension, blood-brain barrier dysfunction, and pulmonary hypertension. It remains unclear whether TMEM16A regulates endothelial angiogenesis, which participates in many physiological and pathological processes. Cholesterol regulates many ion channels including TMEM16A, and high cholesterol levels contribute to endothelial dysfunction. It remains to be determined whether cholesterol regulates TMEM16A expression and function in endothelial cells.
Objective: This study aimed to investigate whether cholesterol regulated TMEM16A expression and function in endothelial angiogenesis.
Methods: Whole-cell patch clamp techniques were used to record Ca2+-activated Cl- currents in human aortic endothelial cells (HAECs) and HEK293 cells transfected with TMEM16A-overexpressing plasmids. Western blot was used to examine the expression of TMEM16A and DNA methyltransferase 1 (DNMT1) in HAECs. CCK-8 assay, would healing assay, and tube formation assay were used to test endothelial cell proliferation, migration and angiogenesis, respectively.
Results: TMEM16A mediates the Ca2+-activated Cl- channel in HAECs. Cholesterol treatment inhibited TMEM16A expression via upregulation of DNMT1 in HAECs, and the inhibitory effect of cholesterol on TMEM16A expression was blocked by 5-aza, the DNMT1 inhibitor. In addition, direct application of cholesterol inhibited TMEM16A currents in heterologous HEK293 cells with an IC50 of 0.1209 μM. Similarly, cholesterol directly inhibited TMEM16A currents in HAECs. Furthermore, TMEM16A knockdown increased in vitro tube formation, cell migration and proliferation of HAECs, and TMEM16A overexpression produced the opposite effect.
Conclusion: This study reveals a novel mechanism of cholesterol-mediated TMEM16A inhibition, by which cholesterol reduces TMEM16A expression via DNMT1-mediated methylation and directly inhibits channel activities. TMEM16A channel inhibition promotes endothelial cell angiogenesis.
| Medienart: |
E-Artikel |
|---|
| Erscheinungsjahr: |
2021 |
|---|---|
| Erschienen: |
2021 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:29 |
|---|---|
| Enthalten in: |
Journal of advanced research - 29(2021) vom: 01. März, Seite 23-32 |
| Sprache: |
Englisch |
|---|
| Beteiligte Personen: |
Ma, Ke [VerfasserIn] |
|---|
| Links: |
|---|
| Anmerkungen: |
Date Completed 24.09.2021 Date Revised 24.09.2021 published: Electronic-eCollection Citation Status MEDLINE |
|---|
| doi: |
10.1016/j.jare.2020.09.003 |
|---|
| funding: |
|
|---|---|
| Förderinstitution / Projekttitel: |
|
| PPN (Katalog-ID): |
NLM323970656 |
|---|
| LEADER | 01000naa a22002652 4500 | ||
|---|---|---|---|
| 001 | NLM323970656 | ||
| 003 | DE-627 | ||
| 005 | 20231225185322.0 | ||
| 007 | cr uuu---uuuuu | ||
| 008 | 231225s2021 xx |||||o 00| ||eng c | ||
| 024 | 7 | |a 10.1016/j.jare.2020.09.003 |2 doi | |
| 028 | 5 | 2 | |a pubmed24n1079.xml |
| 035 | |a (DE-627)NLM323970656 | ||
| 035 | |a (NLM)33842002 | ||
| 040 | |a DE-627 |b ger |c DE-627 |e rakwb | ||
| 041 | |a eng | ||
| 100 | 1 | |a Ma, Ke |e verfasserin |4 aut | |
| 245 | 1 | 0 | |a Ca2+-activated Cl- channel TMEM16A inhibition by cholesterol promotes angiogenesis in endothelial cells |
| 264 | 1 | |c 2021 | |
| 336 | |a Text |b txt |2 rdacontent | ||
| 337 | |a ƒaComputermedien |b c |2 rdamedia | ||
| 338 | |a ƒa Online-Ressource |b cr |2 rdacarrier | ||
| 500 | |a Date Completed 24.09.2021 | ||
| 500 | |a Date Revised 24.09.2021 | ||
| 500 | |a published: Electronic-eCollection | ||
| 500 | |a Citation Status MEDLINE | ||
| 520 | |a © 2021 The Authors. Published by Elsevier B.V. on behalf of Cairo University. | ||
| 520 | |a Introduction: Ca2+-activated Cl- channel TMEM16A is expressed in endothelial cells, and contributes to many diseases such as hypertension, blood-brain barrier dysfunction, and pulmonary hypertension. It remains unclear whether TMEM16A regulates endothelial angiogenesis, which participates in many physiological and pathological processes. Cholesterol regulates many ion channels including TMEM16A, and high cholesterol levels contribute to endothelial dysfunction. It remains to be determined whether cholesterol regulates TMEM16A expression and function in endothelial cells | ||
| 520 | |a Objective: This study aimed to investigate whether cholesterol regulated TMEM16A expression and function in endothelial angiogenesis | ||
| 520 | |a Methods: Whole-cell patch clamp techniques were used to record Ca2+-activated Cl- currents in human aortic endothelial cells (HAECs) and HEK293 cells transfected with TMEM16A-overexpressing plasmids. Western blot was used to examine the expression of TMEM16A and DNA methyltransferase 1 (DNMT1) in HAECs. CCK-8 assay, would healing assay, and tube formation assay were used to test endothelial cell proliferation, migration and angiogenesis, respectively | ||
| 520 | |a Results: TMEM16A mediates the Ca2+-activated Cl- channel in HAECs. Cholesterol treatment inhibited TMEM16A expression via upregulation of DNMT1 in HAECs, and the inhibitory effect of cholesterol on TMEM16A expression was blocked by 5-aza, the DNMT1 inhibitor. In addition, direct application of cholesterol inhibited TMEM16A currents in heterologous HEK293 cells with an IC50 of 0.1209 μM. Similarly, cholesterol directly inhibited TMEM16A currents in HAECs. Furthermore, TMEM16A knockdown increased in vitro tube formation, cell migration and proliferation of HAECs, and TMEM16A overexpression produced the opposite effect | ||
| 520 | |a Conclusion: This study reveals a novel mechanism of cholesterol-mediated TMEM16A inhibition, by which cholesterol reduces TMEM16A expression via DNMT1-mediated methylation and directly inhibits channel activities. TMEM16A channel inhibition promotes endothelial cell angiogenesis | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a Research Support, Non-U.S. Gov't | |
| 650 | 4 | |a 5-aza, 5-Aza-2′-deoxycytidine | |
| 650 | 4 | |a ANOVA, analysis of variance | |
| 650 | 4 | |a Angiogenesis | |
| 650 | 4 | |a CCK-8, Cell Counting Kit-8 | |
| 650 | 4 | |a CaCCs, Ca2+-activated Cl− currents | |
| 650 | 4 | |a Cholesterol | |
| 650 | 4 | |a DMEM, Dulbecco’s Modified Eagle Medium | |
| 650 | 4 | |a DNMT1, DNA methyltransferase 1 | |
| 650 | 4 | |a EGTA, ethylene glycol-bis(2-aminoethyl ether)-N,N,N',N'-tetraacetic acid | |
| 650 | 4 | |a Endothelial cells | |
| 650 | 4 | |a FBS, fetal bovine serum | |
| 650 | 4 | |a HAECs, human aortic endothelial cells | |
| 650 | 4 | |a HEPES, N-2-hydroxyethil-piperazine-N'-2-ethanesulfonic acid | |
| 650 | 4 | |a MβCD, methyl-β cyclodextrin | |
| 650 | 4 | |a NMDG, N-methyl-D-glucamine | |
| 650 | 4 | |a PVDF, polyvinylidene fluoride | |
| 650 | 4 | |a RIPA, radio immunoprecipitation assay | |
| 650 | 4 | |a ROS, reactive oxygen species | |
| 650 | 4 | |a SE, standard error | |
| 650 | 4 | |a TMEM16A | |
| 650 | 4 | |a shRNAs, short hairpin RNAs | |
| 650 | 7 | |a Anoctamin-1 |2 NLM | |
| 650 | 7 | |a Chloride Channels |2 NLM | |
| 650 | 7 | |a Cholesterol |2 NLM | |
| 650 | 7 | |a 97C5T2UQ7J |2 NLM | |
| 650 | 7 | |a DNA (Cytosine-5-)-Methyltransferase 1 |2 NLM | |
| 650 | 7 | |a EC 2.1.1.37 |2 NLM | |
| 650 | 7 | |a DNMT1 protein, human |2 NLM | |
| 650 | 7 | |a EC 2.1.1.37 |2 NLM | |
| 650 | 7 | |a Calcium |2 NLM | |
| 650 | 7 | |a SY7Q814VUP |2 NLM | |
| 700 | 1 | |a Liu, Sitong |e verfasserin |4 aut | |
| 700 | 1 | |a Liang, Hongyue |e verfasserin |4 aut | |
| 700 | 1 | |a Wang, Guan |e verfasserin |4 aut | |
| 700 | 1 | |a Wang, Tianyu |e verfasserin |4 aut | |
| 700 | 1 | |a Luo, Shuya |e verfasserin |4 aut | |
| 700 | 1 | |a Gao, Kuan |e verfasserin |4 aut | |
| 700 | 1 | |a Wang, Hui |e verfasserin |4 aut | |
| 700 | 1 | |a Liu, Mei |e verfasserin |4 aut | |
| 700 | 1 | |a Bai, Lichuan |e verfasserin |4 aut | |
| 700 | 1 | |a Xiao, Qinghuan |e verfasserin |4 aut | |
| 773 | 0 | 8 | |i Enthalten in |t Journal of advanced research |d 2013 |g 29(2021) vom: 01. März, Seite 23-32 |w (DE-627)NLM246273542 |x 2090-1224 |7 nnns |
| 773 | 1 | 8 | |g volume:29 |g year:2021 |g day:01 |g month:03 |g pages:23-32 |
| 856 | 4 | 0 | |u http://dx.doi.org/10.1016/j.jare.2020.09.003 |3 Volltext |
| 912 | |a GBV_USEFLAG_A | ||
| 912 | |a GBV_NLM | ||
| 951 | |a AR | ||
| 952 | |d 29 |j 2021 |b 01 |c 03 |h 23-32 | ||