Arenobufagin induces apoptosis in acute leukemia cells by activating Rho A/ROCK1 signaling pathway

Objective To investigate the apoptosis-inducing effect of arenobufagin (ARE) on acute leukemia cells and explore its possible molecular mechanism. Methods Acute leukemia cell lines Jurkat and MV-4-11 were treated with different concentrations of ARE (0, 10, 20, 40 or 80 nmol/L) for 24 h, or with 80 nmol/L ARE for different durations (0, 6, 12, 18 or 24 h), or with 80 nmol/L ARE for 24 h after being pretreated with 20 μmol/L Y-27632 (inhibitor of ROCK1) for 1 h, respectively. Then, MTT assay and flow cytometry were used to respectively detect the effect of ARE on cell proliferation and apoptosis in acute leukemia cells; Western blotting and co-immunoprecipitation (CoIP) assay were performed to determine the expression of apoptosis-related and Rho A/ROCK1 pathway-related proteins in the acute leukemia cells after ARE treatment. Results ARE inhibited the proliferation and induced apoptosis of acute leukemia cells in dose- and time-dependent manners (P < 0.01). Western blotting results showed that ARE activated ROCK1 signaling pathway, facilitated the translocation of BAX from cytoplasm to mitochondria, promoted the release of mitochondrial proteins (Cytochrome c, AIF), increased the cleavage/activation of Caspase 3 and Caspase 7 as well as the degradation of Poly(ADP-ribose) polymerase 1 (PARP1), and then reduced the expression of X-linked inhibitor of apoptosis (XIAP). The above effects presented dose- and time-dependence (P < 0.01). Interruption of ROCK1 pathway by the pharmacological inhibitor Y-27632 significantly attenuated ARE-mediated mitochondrial translocation of BAX, reduced cytosolic release of Cytochrome c and AIF from mitochondria, suppressed the activation of Caspase 3 and Caspase 7, the degradation of cleavage of PARP1, and down-regulated XIAP expression as well as blocked apoptosis (P < 0.01). Conclusion ARE effectively induces apoptosis of acute leukemia cells (Jurkat, MV-4-11) through the activation of Rho A/ROCK1 signaling pathway. Ausführliche Beschreibung

Gespeichert in:

Autor*in:	LI Zhiqiang [verfasserIn] JIANG Xiuxing [verfasserIn] HU Jinjiao [verfasserIn] DING Xin [verfasserIn] LEI Ling [verfasserIn] GAO Ning [verfasserIn]

Format:	E-Artikel
Sprache:	Chinesisch

Erschienen:	2022

Schlagwörter:	arenobufagin acute leukemia rho-associated kinase 1 apoptosis

Übergeordnetes Werk:	In: Di-san junyi daxue xuebao - Editorial Office of Journal of Third Military Medical University, 2021, 44(2022), 7, Seite 700-710
Übergeordnetes Werk:	volume:44 ; year:2022 ; number:7 ; pages:700-710

Links:	Link aufrufen Link aufrufen Link aufrufen Journal toc

DOI / URN:	10.16016/j.2097-0927.202109162

Katalog-ID:	DOAJ035009594

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520			\|a Objective To investigate the apoptosis-inducing effect of arenobufagin (ARE) on acute leukemia cells and explore its possible molecular mechanism. Methods Acute leukemia cell lines Jurkat and MV-4-11 were treated with different concentrations of ARE (0, 10, 20, 40 or 80 nmol/L) for 24 h, or with 80 nmol/L ARE for different durations (0, 6, 12, 18 or 24 h), or with 80 nmol/L ARE for 24 h after being pretreated with 20 μmol/L Y-27632 (inhibitor of ROCK1) for 1 h, respectively. Then, MTT assay and flow cytometry were used to respectively detect the effect of ARE on cell proliferation and apoptosis in acute leukemia cells; Western blotting and co-immunoprecipitation (CoIP) assay were performed to determine the expression of apoptosis-related and Rho A/ROCK1 pathway-related proteins in the acute leukemia cells after ARE treatment. Results ARE inhibited the proliferation and induced apoptosis of acute leukemia cells in dose- and time-dependent manners (P < 0.01). Western blotting results showed that ARE activated ROCK1 signaling pathway, facilitated the translocation of BAX from cytoplasm to mitochondria, promoted the release of mitochondrial proteins (Cytochrome c, AIF), increased the cleavage/activation of Caspase 3 and Caspase 7 as well as the degradation of Poly(ADP-ribose) polymerase 1 (PARP1), and then reduced the expression of X-linked inhibitor of apoptosis (XIAP). The above effects presented dose- and time-dependence (P < 0.01). Interruption of ROCK1 pathway by the pharmacological inhibitor Y-27632 significantly attenuated ARE-mediated mitochondrial translocation of BAX, reduced cytosolic release of Cytochrome c and AIF from mitochondria, suppressed the activation of Caspase 3 and Caspase 7, the degradation of cleavage of PARP1, and down-regulated XIAP expression as well as blocked apoptosis (P < 0.01). Conclusion ARE effectively induces apoptosis of acute leukemia cells (Jurkat, MV-4-11) through the activation of Rho A/ROCK1 signaling pathway.
650		4	\|a arenobufagin
650		4	\|a acute leukemia
650		4	\|a rho-associated kinase 1
650		4	\|a apoptosis
653		0	\|a Medicine (General)
700	0		\|a JIANG Xiuxing \|e verfasserin \|4 aut
700	0		\|a HU Jinjiao \|e verfasserin \|4 aut
700	0		\|a DING Xin \|e verfasserin \|4 aut
700	0		\|a LEI Ling \|e verfasserin \|4 aut
700	0		\|a GAO Ning \|e verfasserin \|4 aut
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allfields	10.16016/j.2097-0927.202109162 doi (DE-627)DOAJ035009594 (DE-599)DOAJ56854f7b76cf494385a79ba07ba31d02 DE-627 ger DE-627 rakwb chi R5-920 LI Zhiqiang verfasserin aut Arenobufagin induces apoptosis in acute leukemia cells by activating Rho A/ROCK1 signaling pathway 2022 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier Objective To investigate the apoptosis-inducing effect of arenobufagin (ARE) on acute leukemia cells and explore its possible molecular mechanism. Methods Acute leukemia cell lines Jurkat and MV-4-11 were treated with different concentrations of ARE (0, 10, 20, 40 or 80 nmol/L) for 24 h, or with 80 nmol/L ARE for different durations (0, 6, 12, 18 or 24 h), or with 80 nmol/L ARE for 24 h after being pretreated with 20 μmol/L Y-27632 (inhibitor of ROCK1) for 1 h, respectively. Then, MTT assay and flow cytometry were used to respectively detect the effect of ARE on cell proliferation and apoptosis in acute leukemia cells; Western blotting and co-immunoprecipitation (CoIP) assay were performed to determine the expression of apoptosis-related and Rho A/ROCK1 pathway-related proteins in the acute leukemia cells after ARE treatment. Results ARE inhibited the proliferation and induced apoptosis of acute leukemia cells in dose- and time-dependent manners (P < 0.01). Western blotting results showed that ARE activated ROCK1 signaling pathway, facilitated the translocation of BAX from cytoplasm to mitochondria, promoted the release of mitochondrial proteins (Cytochrome c, AIF), increased the cleavage/activation of Caspase 3 and Caspase 7 as well as the degradation of Poly(ADP-ribose) polymerase 1 (PARP1), and then reduced the expression of X-linked inhibitor of apoptosis (XIAP). The above effects presented dose- and time-dependence (P < 0.01). Interruption of ROCK1 pathway by the pharmacological inhibitor Y-27632 significantly attenuated ARE-mediated mitochondrial translocation of BAX, reduced cytosolic release of Cytochrome c and AIF from mitochondria, suppressed the activation of Caspase 3 and Caspase 7, the degradation of cleavage of PARP1, and down-regulated XIAP expression as well as blocked apoptosis (P < 0.01). Conclusion ARE effectively induces apoptosis of acute leukemia cells (Jurkat, MV-4-11) through the activation of Rho A/ROCK1 signaling pathway. arenobufagin acute leukemia rho-associated kinase 1 apoptosis Medicine (General) JIANG Xiuxing verfasserin aut HU Jinjiao verfasserin aut DING Xin verfasserin aut LEI Ling verfasserin aut GAO Ning verfasserin aut In Di-san junyi daxue xuebao Editorial Office of Journal of Third Military Medical University, 2021 44(2022), 7, Seite 700-710 (DE-627)1760645346 10005404 nnns volume:44 year:2022 number:7 pages:700-710 https://doi.org/10.16016/j.2097-0927.202109162 kostenfrei https://doaj.org/article/56854f7b76cf494385a79ba07ba31d02 kostenfrei http://aammt.tmmu.edu.cn/Upload/rhtml/202109162.htm kostenfrei https://doaj.org/toc/1000-5404 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ AR 44 2022 7 700-710
spelling	10.16016/j.2097-0927.202109162 doi (DE-627)DOAJ035009594 (DE-599)DOAJ56854f7b76cf494385a79ba07ba31d02 DE-627 ger DE-627 rakwb chi R5-920 LI Zhiqiang verfasserin aut Arenobufagin induces apoptosis in acute leukemia cells by activating Rho A/ROCK1 signaling pathway 2022 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier Objective To investigate the apoptosis-inducing effect of arenobufagin (ARE) on acute leukemia cells and explore its possible molecular mechanism. Methods Acute leukemia cell lines Jurkat and MV-4-11 were treated with different concentrations of ARE (0, 10, 20, 40 or 80 nmol/L) for 24 h, or with 80 nmol/L ARE for different durations (0, 6, 12, 18 or 24 h), or with 80 nmol/L ARE for 24 h after being pretreated with 20 μmol/L Y-27632 (inhibitor of ROCK1) for 1 h, respectively. Then, MTT assay and flow cytometry were used to respectively detect the effect of ARE on cell proliferation and apoptosis in acute leukemia cells; Western blotting and co-immunoprecipitation (CoIP) assay were performed to determine the expression of apoptosis-related and Rho A/ROCK1 pathway-related proteins in the acute leukemia cells after ARE treatment. Results ARE inhibited the proliferation and induced apoptosis of acute leukemia cells in dose- and time-dependent manners (P < 0.01). Western blotting results showed that ARE activated ROCK1 signaling pathway, facilitated the translocation of BAX from cytoplasm to mitochondria, promoted the release of mitochondrial proteins (Cytochrome c, AIF), increased the cleavage/activation of Caspase 3 and Caspase 7 as well as the degradation of Poly(ADP-ribose) polymerase 1 (PARP1), and then reduced the expression of X-linked inhibitor of apoptosis (XIAP). The above effects presented dose- and time-dependence (P < 0.01). Interruption of ROCK1 pathway by the pharmacological inhibitor Y-27632 significantly attenuated ARE-mediated mitochondrial translocation of BAX, reduced cytosolic release of Cytochrome c and AIF from mitochondria, suppressed the activation of Caspase 3 and Caspase 7, the degradation of cleavage of PARP1, and down-regulated XIAP expression as well as blocked apoptosis (P < 0.01). Conclusion ARE effectively induces apoptosis of acute leukemia cells (Jurkat, MV-4-11) through the activation of Rho A/ROCK1 signaling pathway. arenobufagin acute leukemia rho-associated kinase 1 apoptosis Medicine (General) JIANG Xiuxing verfasserin aut HU Jinjiao verfasserin aut DING Xin verfasserin aut LEI Ling verfasserin aut GAO Ning verfasserin aut In Di-san junyi daxue xuebao Editorial Office of Journal of Third Military Medical University, 2021 44(2022), 7, Seite 700-710 (DE-627)1760645346 10005404 nnns volume:44 year:2022 number:7 pages:700-710 https://doi.org/10.16016/j.2097-0927.202109162 kostenfrei https://doaj.org/article/56854f7b76cf494385a79ba07ba31d02 kostenfrei http://aammt.tmmu.edu.cn/Upload/rhtml/202109162.htm kostenfrei https://doaj.org/toc/1000-5404 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ AR 44 2022 7 700-710
allfields_unstemmed	10.16016/j.2097-0927.202109162 doi (DE-627)DOAJ035009594 (DE-599)DOAJ56854f7b76cf494385a79ba07ba31d02 DE-627 ger DE-627 rakwb chi R5-920 LI Zhiqiang verfasserin aut Arenobufagin induces apoptosis in acute leukemia cells by activating Rho A/ROCK1 signaling pathway 2022 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier Objective To investigate the apoptosis-inducing effect of arenobufagin (ARE) on acute leukemia cells and explore its possible molecular mechanism. Methods Acute leukemia cell lines Jurkat and MV-4-11 were treated with different concentrations of ARE (0, 10, 20, 40 or 80 nmol/L) for 24 h, or with 80 nmol/L ARE for different durations (0, 6, 12, 18 or 24 h), or with 80 nmol/L ARE for 24 h after being pretreated with 20 μmol/L Y-27632 (inhibitor of ROCK1) for 1 h, respectively. Then, MTT assay and flow cytometry were used to respectively detect the effect of ARE on cell proliferation and apoptosis in acute leukemia cells; Western blotting and co-immunoprecipitation (CoIP) assay were performed to determine the expression of apoptosis-related and Rho A/ROCK1 pathway-related proteins in the acute leukemia cells after ARE treatment. Results ARE inhibited the proliferation and induced apoptosis of acute leukemia cells in dose- and time-dependent manners (P < 0.01). Western blotting results showed that ARE activated ROCK1 signaling pathway, facilitated the translocation of BAX from cytoplasm to mitochondria, promoted the release of mitochondrial proteins (Cytochrome c, AIF), increased the cleavage/activation of Caspase 3 and Caspase 7 as well as the degradation of Poly(ADP-ribose) polymerase 1 (PARP1), and then reduced the expression of X-linked inhibitor of apoptosis (XIAP). The above effects presented dose- and time-dependence (P < 0.01). Interruption of ROCK1 pathway by the pharmacological inhibitor Y-27632 significantly attenuated ARE-mediated mitochondrial translocation of BAX, reduced cytosolic release of Cytochrome c and AIF from mitochondria, suppressed the activation of Caspase 3 and Caspase 7, the degradation of cleavage of PARP1, and down-regulated XIAP expression as well as blocked apoptosis (P < 0.01). Conclusion ARE effectively induces apoptosis of acute leukemia cells (Jurkat, MV-4-11) through the activation of Rho A/ROCK1 signaling pathway. arenobufagin acute leukemia rho-associated kinase 1 apoptosis Medicine (General) JIANG Xiuxing verfasserin aut HU Jinjiao verfasserin aut DING Xin verfasserin aut LEI Ling verfasserin aut GAO Ning verfasserin aut In Di-san junyi daxue xuebao Editorial Office of Journal of Third Military Medical University, 2021 44(2022), 7, Seite 700-710 (DE-627)1760645346 10005404 nnns volume:44 year:2022 number:7 pages:700-710 https://doi.org/10.16016/j.2097-0927.202109162 kostenfrei https://doaj.org/article/56854f7b76cf494385a79ba07ba31d02 kostenfrei http://aammt.tmmu.edu.cn/Upload/rhtml/202109162.htm kostenfrei https://doaj.org/toc/1000-5404 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ AR 44 2022 7 700-710
allfieldsGer	10.16016/j.2097-0927.202109162 doi (DE-627)DOAJ035009594 (DE-599)DOAJ56854f7b76cf494385a79ba07ba31d02 DE-627 ger DE-627 rakwb chi R5-920 LI Zhiqiang verfasserin aut Arenobufagin induces apoptosis in acute leukemia cells by activating Rho A/ROCK1 signaling pathway 2022 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier Objective To investigate the apoptosis-inducing effect of arenobufagin (ARE) on acute leukemia cells and explore its possible molecular mechanism. Methods Acute leukemia cell lines Jurkat and MV-4-11 were treated with different concentrations of ARE (0, 10, 20, 40 or 80 nmol/L) for 24 h, or with 80 nmol/L ARE for different durations (0, 6, 12, 18 or 24 h), or with 80 nmol/L ARE for 24 h after being pretreated with 20 μmol/L Y-27632 (inhibitor of ROCK1) for 1 h, respectively. Then, MTT assay and flow cytometry were used to respectively detect the effect of ARE on cell proliferation and apoptosis in acute leukemia cells; Western blotting and co-immunoprecipitation (CoIP) assay were performed to determine the expression of apoptosis-related and Rho A/ROCK1 pathway-related proteins in the acute leukemia cells after ARE treatment. Results ARE inhibited the proliferation and induced apoptosis of acute leukemia cells in dose- and time-dependent manners (P < 0.01). Western blotting results showed that ARE activated ROCK1 signaling pathway, facilitated the translocation of BAX from cytoplasm to mitochondria, promoted the release of mitochondrial proteins (Cytochrome c, AIF), increased the cleavage/activation of Caspase 3 and Caspase 7 as well as the degradation of Poly(ADP-ribose) polymerase 1 (PARP1), and then reduced the expression of X-linked inhibitor of apoptosis (XIAP). The above effects presented dose- and time-dependence (P < 0.01). Interruption of ROCK1 pathway by the pharmacological inhibitor Y-27632 significantly attenuated ARE-mediated mitochondrial translocation of BAX, reduced cytosolic release of Cytochrome c and AIF from mitochondria, suppressed the activation of Caspase 3 and Caspase 7, the degradation of cleavage of PARP1, and down-regulated XIAP expression as well as blocked apoptosis (P < 0.01). Conclusion ARE effectively induces apoptosis of acute leukemia cells (Jurkat, MV-4-11) through the activation of Rho A/ROCK1 signaling pathway. arenobufagin acute leukemia rho-associated kinase 1 apoptosis Medicine (General) JIANG Xiuxing verfasserin aut HU Jinjiao verfasserin aut DING Xin verfasserin aut LEI Ling verfasserin aut GAO Ning verfasserin aut In Di-san junyi daxue xuebao Editorial Office of Journal of Third Military Medical University, 2021 44(2022), 7, Seite 700-710 (DE-627)1760645346 10005404 nnns volume:44 year:2022 number:7 pages:700-710 https://doi.org/10.16016/j.2097-0927.202109162 kostenfrei https://doaj.org/article/56854f7b76cf494385a79ba07ba31d02 kostenfrei http://aammt.tmmu.edu.cn/Upload/rhtml/202109162.htm kostenfrei https://doaj.org/toc/1000-5404 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ AR 44 2022 7 700-710
allfieldsSound	10.16016/j.2097-0927.202109162 doi (DE-627)DOAJ035009594 (DE-599)DOAJ56854f7b76cf494385a79ba07ba31d02 DE-627 ger DE-627 rakwb chi R5-920 LI Zhiqiang verfasserin aut Arenobufagin induces apoptosis in acute leukemia cells by activating Rho A/ROCK1 signaling pathway 2022 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier Objective To investigate the apoptosis-inducing effect of arenobufagin (ARE) on acute leukemia cells and explore its possible molecular mechanism. Methods Acute leukemia cell lines Jurkat and MV-4-11 were treated with different concentrations of ARE (0, 10, 20, 40 or 80 nmol/L) for 24 h, or with 80 nmol/L ARE for different durations (0, 6, 12, 18 or 24 h), or with 80 nmol/L ARE for 24 h after being pretreated with 20 μmol/L Y-27632 (inhibitor of ROCK1) for 1 h, respectively. Then, MTT assay and flow cytometry were used to respectively detect the effect of ARE on cell proliferation and apoptosis in acute leukemia cells; Western blotting and co-immunoprecipitation (CoIP) assay were performed to determine the expression of apoptosis-related and Rho A/ROCK1 pathway-related proteins in the acute leukemia cells after ARE treatment. Results ARE inhibited the proliferation and induced apoptosis of acute leukemia cells in dose- and time-dependent manners (P < 0.01). Western blotting results showed that ARE activated ROCK1 signaling pathway, facilitated the translocation of BAX from cytoplasm to mitochondria, promoted the release of mitochondrial proteins (Cytochrome c, AIF), increased the cleavage/activation of Caspase 3 and Caspase 7 as well as the degradation of Poly(ADP-ribose) polymerase 1 (PARP1), and then reduced the expression of X-linked inhibitor of apoptosis (XIAP). The above effects presented dose- and time-dependence (P < 0.01). Interruption of ROCK1 pathway by the pharmacological inhibitor Y-27632 significantly attenuated ARE-mediated mitochondrial translocation of BAX, reduced cytosolic release of Cytochrome c and AIF from mitochondria, suppressed the activation of Caspase 3 and Caspase 7, the degradation of cleavage of PARP1, and down-regulated XIAP expression as well as blocked apoptosis (P < 0.01). Conclusion ARE effectively induces apoptosis of acute leukemia cells (Jurkat, MV-4-11) through the activation of Rho A/ROCK1 signaling pathway. arenobufagin acute leukemia rho-associated kinase 1 apoptosis Medicine (General) JIANG Xiuxing verfasserin aut HU Jinjiao verfasserin aut DING Xin verfasserin aut LEI Ling verfasserin aut GAO Ning verfasserin aut In Di-san junyi daxue xuebao Editorial Office of Journal of Third Military Medical University, 2021 44(2022), 7, Seite 700-710 (DE-627)1760645346 10005404 nnns volume:44 year:2022 number:7 pages:700-710 https://doi.org/10.16016/j.2097-0927.202109162 kostenfrei https://doaj.org/article/56854f7b76cf494385a79ba07ba31d02 kostenfrei http://aammt.tmmu.edu.cn/Upload/rhtml/202109162.htm kostenfrei https://doaj.org/toc/1000-5404 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ AR 44 2022 7 700-710
language	Chinese
source	In Di-san junyi daxue xuebao 44(2022), 7, Seite 700-710 volume:44 year:2022 number:7 pages:700-710
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container_title	Di-san junyi daxue xuebao
authorswithroles_txt_mv	LI Zhiqiang @@aut@@ JIANG Xiuxing @@aut@@ HU Jinjiao @@aut@@ DING Xin @@aut@@ LEI Ling @@aut@@ GAO Ning @@aut@@
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Methods Acute leukemia cell lines Jurkat and MV-4-11 were treated with different concentrations of ARE (0, 10, 20, 40 or 80 nmol/L) for 24 h, or with 80 nmol/L ARE for different durations (0, 6, 12, 18 or 24 h), or with 80 nmol/L ARE for 24 h after being pretreated with 20 μmol/L Y-27632 (inhibitor of ROCK1) for 1 h, respectively. Then, MTT assay and flow cytometry were used to respectively detect the effect of ARE on cell proliferation and apoptosis in acute leukemia cells; Western blotting and co-immunoprecipitation (CoIP) assay were performed to determine the expression of apoptosis-related and Rho A/ROCK1 pathway-related proteins in the acute leukemia cells after ARE treatment. Results ARE inhibited the proliferation and induced apoptosis of acute leukemia cells in dose- and time-dependent manners (P < 0.01). Western blotting results showed that ARE activated ROCK1 signaling pathway, facilitated the translocation of BAX from cytoplasm to mitochondria, promoted the release of mitochondrial proteins (Cytochrome c, AIF), increased the cleavage/activation of Caspase 3 and Caspase 7 as well as the degradation of Poly(ADP-ribose) polymerase 1 (PARP1), and then reduced the expression of X-linked inhibitor of apoptosis (XIAP). The above effects presented dose- and time-dependence (P < 0.01). Interruption of ROCK1 pathway by the pharmacological inhibitor Y-27632 significantly attenuated ARE-mediated mitochondrial translocation of BAX, reduced cytosolic release of Cytochrome c and AIF from mitochondria, suppressed the activation of Caspase 3 and Caspase 7, the degradation of cleavage of PARP1, and down-regulated XIAP expression as well as blocked apoptosis (P < 0.01). 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callnumber-first	R - Medicine
author	LI Zhiqiang
spellingShingle	LI Zhiqiang misc R5-920 misc arenobufagin misc acute leukemia misc rho-associated kinase 1 misc apoptosis misc Medicine (General) Arenobufagin induces apoptosis in acute leukemia cells by activating Rho A/ROCK1 signaling pathway
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topic_title	R5-920 Arenobufagin induces apoptosis in acute leukemia cells by activating Rho A/ROCK1 signaling pathway arenobufagin acute leukemia rho-associated kinase 1 apoptosis
topic	misc R5-920 misc arenobufagin misc acute leukemia misc rho-associated kinase 1 misc apoptosis misc Medicine (General)
topic_unstemmed	misc R5-920 misc arenobufagin misc acute leukemia misc rho-associated kinase 1 misc apoptosis misc Medicine (General)
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title	Arenobufagin induces apoptosis in acute leukemia cells by activating Rho A/ROCK1 signaling pathway
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title_full	Arenobufagin induces apoptosis in acute leukemia cells by activating Rho A/ROCK1 signaling pathway
author_sort	LI Zhiqiang
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author_browse	LI Zhiqiang JIANG Xiuxing HU Jinjiao DING Xin LEI Ling GAO Ning
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title_sort	arenobufagin induces apoptosis in acute leukemia cells by activating rho a/rock1 signaling pathway
callnumber	R5-920
title_auth	Arenobufagin induces apoptosis in acute leukemia cells by activating Rho A/ROCK1 signaling pathway
abstract	Objective To investigate the apoptosis-inducing effect of arenobufagin (ARE) on acute leukemia cells and explore its possible molecular mechanism. Methods Acute leukemia cell lines Jurkat and MV-4-11 were treated with different concentrations of ARE (0, 10, 20, 40 or 80 nmol/L) for 24 h, or with 80 nmol/L ARE for different durations (0, 6, 12, 18 or 24 h), or with 80 nmol/L ARE for 24 h after being pretreated with 20 μmol/L Y-27632 (inhibitor of ROCK1) for 1 h, respectively. Then, MTT assay and flow cytometry were used to respectively detect the effect of ARE on cell proliferation and apoptosis in acute leukemia cells; Western blotting and co-immunoprecipitation (CoIP) assay were performed to determine the expression of apoptosis-related and Rho A/ROCK1 pathway-related proteins in the acute leukemia cells after ARE treatment. Results ARE inhibited the proliferation and induced apoptosis of acute leukemia cells in dose- and time-dependent manners (P < 0.01). Western blotting results showed that ARE activated ROCK1 signaling pathway, facilitated the translocation of BAX from cytoplasm to mitochondria, promoted the release of mitochondrial proteins (Cytochrome c, AIF), increased the cleavage/activation of Caspase 3 and Caspase 7 as well as the degradation of Poly(ADP-ribose) polymerase 1 (PARP1), and then reduced the expression of X-linked inhibitor of apoptosis (XIAP). The above effects presented dose- and time-dependence (P < 0.01). Interruption of ROCK1 pathway by the pharmacological inhibitor Y-27632 significantly attenuated ARE-mediated mitochondrial translocation of BAX, reduced cytosolic release of Cytochrome c and AIF from mitochondria, suppressed the activation of Caspase 3 and Caspase 7, the degradation of cleavage of PARP1, and down-regulated XIAP expression as well as blocked apoptosis (P < 0.01). Conclusion ARE effectively induces apoptosis of acute leukemia cells (Jurkat, MV-4-11) through the activation of Rho A/ROCK1 signaling pathway.
abstractGer	Objective To investigate the apoptosis-inducing effect of arenobufagin (ARE) on acute leukemia cells and explore its possible molecular mechanism. Methods Acute leukemia cell lines Jurkat and MV-4-11 were treated with different concentrations of ARE (0, 10, 20, 40 or 80 nmol/L) for 24 h, or with 80 nmol/L ARE for different durations (0, 6, 12, 18 or 24 h), or with 80 nmol/L ARE for 24 h after being pretreated with 20 μmol/L Y-27632 (inhibitor of ROCK1) for 1 h, respectively. Then, MTT assay and flow cytometry were used to respectively detect the effect of ARE on cell proliferation and apoptosis in acute leukemia cells; Western blotting and co-immunoprecipitation (CoIP) assay were performed to determine the expression of apoptosis-related and Rho A/ROCK1 pathway-related proteins in the acute leukemia cells after ARE treatment. Results ARE inhibited the proliferation and induced apoptosis of acute leukemia cells in dose- and time-dependent manners (P < 0.01). Western blotting results showed that ARE activated ROCK1 signaling pathway, facilitated the translocation of BAX from cytoplasm to mitochondria, promoted the release of mitochondrial proteins (Cytochrome c, AIF), increased the cleavage/activation of Caspase 3 and Caspase 7 as well as the degradation of Poly(ADP-ribose) polymerase 1 (PARP1), and then reduced the expression of X-linked inhibitor of apoptosis (XIAP). The above effects presented dose- and time-dependence (P < 0.01). Interruption of ROCK1 pathway by the pharmacological inhibitor Y-27632 significantly attenuated ARE-mediated mitochondrial translocation of BAX, reduced cytosolic release of Cytochrome c and AIF from mitochondria, suppressed the activation of Caspase 3 and Caspase 7, the degradation of cleavage of PARP1, and down-regulated XIAP expression as well as blocked apoptosis (P < 0.01). Conclusion ARE effectively induces apoptosis of acute leukemia cells (Jurkat, MV-4-11) through the activation of Rho A/ROCK1 signaling pathway.
abstract_unstemmed	Objective To investigate the apoptosis-inducing effect of arenobufagin (ARE) on acute leukemia cells and explore its possible molecular mechanism. Methods Acute leukemia cell lines Jurkat and MV-4-11 were treated with different concentrations of ARE (0, 10, 20, 40 or 80 nmol/L) for 24 h, or with 80 nmol/L ARE for different durations (0, 6, 12, 18 or 24 h), or with 80 nmol/L ARE for 24 h after being pretreated with 20 μmol/L Y-27632 (inhibitor of ROCK1) for 1 h, respectively. Then, MTT assay and flow cytometry were used to respectively detect the effect of ARE on cell proliferation and apoptosis in acute leukemia cells; Western blotting and co-immunoprecipitation (CoIP) assay were performed to determine the expression of apoptosis-related and Rho A/ROCK1 pathway-related proteins in the acute leukemia cells after ARE treatment. Results ARE inhibited the proliferation and induced apoptosis of acute leukemia cells in dose- and time-dependent manners (P < 0.01). Western blotting results showed that ARE activated ROCK1 signaling pathway, facilitated the translocation of BAX from cytoplasm to mitochondria, promoted the release of mitochondrial proteins (Cytochrome c, AIF), increased the cleavage/activation of Caspase 3 and Caspase 7 as well as the degradation of Poly(ADP-ribose) polymerase 1 (PARP1), and then reduced the expression of X-linked inhibitor of apoptosis (XIAP). The above effects presented dose- and time-dependence (P < 0.01). Interruption of ROCK1 pathway by the pharmacological inhibitor Y-27632 significantly attenuated ARE-mediated mitochondrial translocation of BAX, reduced cytosolic release of Cytochrome c and AIF from mitochondria, suppressed the activation of Caspase 3 and Caspase 7, the degradation of cleavage of PARP1, and down-regulated XIAP expression as well as blocked apoptosis (P < 0.01). Conclusion ARE effectively induces apoptosis of acute leukemia cells (Jurkat, MV-4-11) through the activation of Rho A/ROCK1 signaling pathway.
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title_short	Arenobufagin induces apoptosis in acute leukemia cells by activating Rho A/ROCK1 signaling pathway
url	https://doi.org/10.16016/j.2097-0927.202109162 https://doaj.org/article/56854f7b76cf494385a79ba07ba31d02 http://aammt.tmmu.edu.cn/Upload/rhtml/202109162.htm https://doaj.org/toc/1000-5404
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Methods Acute leukemia cell lines Jurkat and MV-4-11 were treated with different concentrations of ARE (0, 10, 20, 40 or 80 nmol/L) for 24 h, or with 80 nmol/L ARE for different durations (0, 6, 12, 18 or 24 h), or with 80 nmol/L ARE for 24 h after being pretreated with 20 μmol/L Y-27632 (inhibitor of ROCK1) for 1 h, respectively. Then, MTT assay and flow cytometry were used to respectively detect the effect of ARE on cell proliferation and apoptosis in acute leukemia cells; Western blotting and co-immunoprecipitation (CoIP) assay were performed to determine the expression of apoptosis-related and Rho A/ROCK1 pathway-related proteins in the acute leukemia cells after ARE treatment. Results ARE inhibited the proliferation and induced apoptosis of acute leukemia cells in dose- and time-dependent manners (P < 0.01). 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