Rho/Rhotekin-mediated NF-κB activation confers resistance to apoptosis

Ching-Ann Liu, Mei-Jung Wang, Chin-Wen Chi, Chew-Wun Wu, Jeou-Yuan Chen

Research output: Contribution to journalArticle

68 Citations (Scopus)

Abstract

Rhotekin (RTKN), the gene coding for the Rho effector, RTKN, was shown to be overexpressed in human gastric cancer (GC). In this study, we further showed that RTKN is expressed at a low level in normal cells and is overexpressed in many cancer-derived cell lines. The function of RTKN as an effector protein in Rho GTPase-mediated pathways regulating apoptosis was investigated. By transfection and expression of RTKN in cells that expressed endogenous RTKN at a low basal level, we showed that RTKN overexpression conferred cell resistance to apoptosis induced by serum deprivation or treatment with sodium butyrate, and the increased resistance correlated to the level of RTKN. Conversely, reducing RTKN expression by small interfering RNAs greatly sensitized cells to apoptosis. The RTKN-mediated antiapoptotic effect was blocked by the nuclear factor-κB (NF-κB) inhibitors, curcumin or parthenolide, but not by the phosphatidylinositol 3′-OH-kinase inhibitor, LY294002, or the MAP kinase inhibitor, PD98059. Reporter gene assays and electrophoretic mobility shift assay confirmed that RTKN overexpression led to constitutive activation of NF-κB through the phosphorylation of IκB by IKKβ. By using the RTKN truncation mutants, we showed that RTKN mediated Rho activity eliciting signaling pathway to activate NF-κB, with a concomitant induction of expression of the NF-κB antiapoptotic genes, cIAP-2, BCl-xL, A1, and A20. Consistent with these data, RTKN-expressing cells showed increased chemoresistance to 5-fluorouracil and paclitaxol, and the resistance was greatly attenuated by NF-κB inhibitor. In conclusion, overactivated Rho/ RTKN/NF-κB signaling pathway through overexpression of RTKN may play a key role in gastric tumorigenesis by conferring cells resistance to apoptosis, and this signaling pathway may serve as an important target for novel therapeutic approaches to the treatment of human GC.
Original languageEnglish
Pages (from-to)8731-8742
Number of pages12
JournalOncogene
Volume23
Issue number54
DOIs
Publication statusPublished - 2004
Externally publishedYes

Fingerprint

Apoptosis
Stomach Neoplasms
Phosphatidylinositol 3-Kinase
rho GTP-Binding Proteins
2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one
Curcumin
Butyric Acid
Electrophoretic Mobility Shift Assay
Reporter Genes
Fluorouracil
Small Interfering RNA
Genes
Transfection
Stomach
Carcinogenesis
Phosphotransferases
Phosphorylation
Cell Line
Serum
Neoplasms

Keywords

  • Antiapoptosis
  • Gastric cancer
  • Nuclear factor-κB
  • Rho GTPases
  • Rhotekin
  • 2 (2 amino 3 methoxyphenyl)chromone
  • 2 morpholino 8 phenylchromone
  • butyric acid
  • curcumin
  • fluorouracil
  • I kappa B
  • immunoglobulin enhancer binding protein
  • mitogen activated protein kinase inhibitor
  • paclitaxel
  • parthenolide
  • phosphatidylinositol 3 kinase inhibitor
  • protein
  • Rho factor
  • Rho guanine nucleotide binding protein
  • rhotekin
  • small interfering RNA
  • unclassified drug
  • A1 gene
  • a20 gene
  • apoptosis
  • article
  • assay
  • bcl xl gene
  • cancer cell culture
  • ciap 2 gene
  • controlled study
  • correlation analysis
  • gel mobility shift assay
  • gene
  • gene expression
  • gene function
  • gene induction
  • gene overexpression
  • genetic transfection
  • human
  • human cell
  • mutant
  • nucleotide sequence
  • priority journal
  • protein phosphorylation
  • regulatory mechanism
  • reporter gene
  • rhotekin gene
  • serum
  • signal transduction
  • Amino Acid Sequence
  • Apoptosis
  • Base Sequence
  • Cell Line, Tumor
  • DNA Primers
  • Electrophoretic Mobility Shift Assay
  • Flow Cytometry
  • Humans
  • Intracellular Signaling Peptides and Proteins
  • NF-kappa B
  • Reverse Transcriptase Polymerase Chain Reaction
  • rho GTP-Binding Proteins

Cite this

Liu, C-A., Wang, M-J., Chi, C-W., Wu, C-W., & Chen, J-Y. (2004). Rho/Rhotekin-mediated NF-κB activation confers resistance to apoptosis. Oncogene, 23(54), 8731-8742. https://doi.org/10.1038/sj.onc.1208106

Rho/Rhotekin-mediated NF-κB activation confers resistance to apoptosis. / Liu, Ching-Ann; Wang, Mei-Jung; Chi, Chin-Wen; Wu, Chew-Wun; Chen, Jeou-Yuan.

In: Oncogene, Vol. 23, No. 54, 2004, p. 8731-8742.

Research output: Contribution to journalArticle

Liu, C-A, Wang, M-J, Chi, C-W, Wu, C-W & Chen, J-Y 2004, 'Rho/Rhotekin-mediated NF-κB activation confers resistance to apoptosis', Oncogene, vol. 23, no. 54, pp. 8731-8742. https://doi.org/10.1038/sj.onc.1208106
Liu, Ching-Ann ; Wang, Mei-Jung ; Chi, Chin-Wen ; Wu, Chew-Wun ; Chen, Jeou-Yuan. / Rho/Rhotekin-mediated NF-κB activation confers resistance to apoptosis. In: Oncogene. 2004 ; Vol. 23, No. 54. pp. 8731-8742.
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title = "Rho/Rhotekin-mediated NF-κB activation confers resistance to apoptosis",
abstract = "Rhotekin (RTKN), the gene coding for the Rho effector, RTKN, was shown to be overexpressed in human gastric cancer (GC). In this study, we further showed that RTKN is expressed at a low level in normal cells and is overexpressed in many cancer-derived cell lines. The function of RTKN as an effector protein in Rho GTPase-mediated pathways regulating apoptosis was investigated. By transfection and expression of RTKN in cells that expressed endogenous RTKN at a low basal level, we showed that RTKN overexpression conferred cell resistance to apoptosis induced by serum deprivation or treatment with sodium butyrate, and the increased resistance correlated to the level of RTKN. Conversely, reducing RTKN expression by small interfering RNAs greatly sensitized cells to apoptosis. The RTKN-mediated antiapoptotic effect was blocked by the nuclear factor-κB (NF-κB) inhibitors, curcumin or parthenolide, but not by the phosphatidylinositol 3′-OH-kinase inhibitor, LY294002, or the MAP kinase inhibitor, PD98059. Reporter gene assays and electrophoretic mobility shift assay confirmed that RTKN overexpression led to constitutive activation of NF-κB through the phosphorylation of IκB by IKKβ. By using the RTKN truncation mutants, we showed that RTKN mediated Rho activity eliciting signaling pathway to activate NF-κB, with a concomitant induction of expression of the NF-κB antiapoptotic genes, cIAP-2, BCl-xL, A1, and A20. Consistent with these data, RTKN-expressing cells showed increased chemoresistance to 5-fluorouracil and paclitaxol, and the resistance was greatly attenuated by NF-κB inhibitor. In conclusion, overactivated Rho/ RTKN/NF-κB signaling pathway through overexpression of RTKN may play a key role in gastric tumorigenesis by conferring cells resistance to apoptosis, and this signaling pathway may serve as an important target for novel therapeutic approaches to the treatment of human GC.",
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author = "Ching-Ann Liu and Mei-Jung Wang and Chin-Wen Chi and Chew-Wun Wu and Jeou-Yuan Chen",
note = "被引用次數:52 Export Date: 28 March 2016 CODEN: ONCNE 通訊地址: Chen, J.-Y.; Institute of Biomedical Sciences, Academia Sinica, 128 Section 2 Academia Road, Taipei 11529, Taiwan; 電子郵件: bmchen@ibms.sinica.edu.tw 分析序列編號: GENBANK: BC004558, NM_001165, NM_002046, NM_004049, NM_006290, NM_053056, NM_138578; 化學物質/CAS: 2 (2 amino 3 methoxyphenyl)chromone, 167869-21-8; 2 morpholino 8 phenylchromone, 154447-36-6; butyric acid, 107-92-6, 156-54-7, 461-55-2; curcumin, 458-37-7; fluorouracil, 51-21-8; paclitaxel, 33069-62-4; parthenolide, 20554-84-1; protein, 67254-75-5; DNA Primers; Intracellular Signaling Peptides and Proteins; NF-kappa B; rho GTP-Binding Proteins, EC 3.6.5.2; RTKN protein, human 商標: ly 294002, Sigma, United States; pd 98059, Sigma, United States 製造商: Sigma, United States 參考文獻: Arlt, A., Vorndamm, J., Muerkoster, S., Yu, H., Schmidt, W.E., Folsch, U.R., Schafer, H., (2002) Cancer Res., 62, pp. 910-916; Asin, S., Taylor, J.A., Trushin, S., Bren, G., Paya, C.V., (1999) J. Virol., 73, pp. 3893-3903; Bhat-Nakshatri, P., Sweeney, C.J., Nakshatri, H., (2002) Oncogene, 21, pp. 2066-2078; Bishop, A.L., Hall, A., (2000) Biochem. J., 348, pp. 241-255; Cammarano, M.S., Minden, A., (2001) J. Biol. Chem., 276, pp. 25876-25882; Cao, Y., Bonizzi, G., Seagroves, T.N., Greten, F.R., Johnson, R., Schmidt, E.V., Karin, M., (2001) Cell, 107, pp. 763-775; Chen, J.-Y., Funk, W.D., Wright, W.E., Shay, J.W., Minna, J.D., (1993) Oncogene, 8, pp. 2159-2166; Chiao, P.J., Na, R., Niu, J., Sclabas, G.M., Dong, Q.G., Curley, S.A., (2002) Cancer, 95, pp. 1696-1705; Chuang, S.-E., Yeh, P.-Y., Lu, Y.-S., Lai, G.-M., Liao, C.-M., Gao, M., Cheng, A.-L., (2002) Biochem. Pharmacol., 63, pp. 1709-1716; DiDonato, J.A., Hayakawa, M., Rothwarf, D.M., Zandi, E., Karin, M., (1997) Nature, 388, pp. 548-554; Engers, R., Zwaka, T.P., Gohr, L., Weber, A., Gerharz, C.D., Gabbert, H.E., (2000) Int. J. Cancer, 88, pp. 369-376; Fritz, G., Brachetti, C., Schmidt, M., Kaina, B., (2002) Br. J. Cancer, 87, pp. 635-644; Fritz, G., Just, I., Kaina, B., (1999) Int. J. Cancer, 81, pp. 682-687; Gildea, J.J., Seraj, M.J., Oxford, G., Harding, M.A., Hampton, G.M., Moskaluk, C.A., Frierson, H.F., Theodorescu, D., (2002) Cancer Res., 62, pp. 6418-6423; Ikeda, H., Nagashima, K., Yanase, M., Tomiya, T., Arai, M., Inoue, Y., Tejima, K., Fujiwara, K., (2003) Am. J. Physiol. Gastrointest. Liver Physiol. On-line; Ishizaki, T., Fujisawa, K., Maekawa, M., Watanabe, N., Saito, Y., Narumiya, S., (1997) FEBS Lett., 404, pp. 118-124; Kamai, T., Arai, S., Sumi, T., Tsujii, M., Honda, T., Yamanishi, T., Yoshida, K.I., (2003) BJU Int., 89, pp. 449-453; Kanai, M., Konda, Y., Nakajima, T., Izumi, Y., Takeuchi, T., Chiba, T., (2001) Gastroenterology, 121, pp. 56-67; Kaneko, K., Satoh, K., Masamune, A., Satoh, A., Shimosegawa, T., (2002) Pancreas, 24, pp. 251-257; Karin, M., Cao, Y., Greten, F.R., Li, Z.W., (2002) Nat. Rev. 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year = "2004",
doi = "10.1038/sj.onc.1208106",
language = "English",
volume = "23",
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journal = "Oncogene",
issn = "0950-9232",
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}

TY - JOUR

T1 - Rho/Rhotekin-mediated NF-κB activation confers resistance to apoptosis

AU - Liu, Ching-Ann

AU - Wang, Mei-Jung

AU - Chi, Chin-Wen

AU - Wu, Chew-Wun

AU - Chen, Jeou-Yuan

N1 - 被引用次數:52 Export Date: 28 March 2016 CODEN: ONCNE 通訊地址: Chen, J.-Y.; Institute of Biomedical Sciences, Academia Sinica, 128 Section 2 Academia Road, Taipei 11529, Taiwan; 電子郵件: bmchen@ibms.sinica.edu.tw 分析序列編號: GENBANK: BC004558, NM_001165, NM_002046, NM_004049, NM_006290, NM_053056, NM_138578; 化學物質/CAS: 2 (2 amino 3 methoxyphenyl)chromone, 167869-21-8; 2 morpholino 8 phenylchromone, 154447-36-6; butyric acid, 107-92-6, 156-54-7, 461-55-2; curcumin, 458-37-7; fluorouracil, 51-21-8; paclitaxel, 33069-62-4; parthenolide, 20554-84-1; protein, 67254-75-5; DNA Primers; Intracellular Signaling Peptides and Proteins; NF-kappa B; rho GTP-Binding Proteins, EC 3.6.5.2; RTKN protein, human 商標: ly 294002, Sigma, United States; pd 98059, Sigma, United States 製造商: Sigma, United States 參考文獻: Arlt, A., Vorndamm, J., Muerkoster, S., Yu, H., Schmidt, W.E., Folsch, U.R., Schafer, H., (2002) Cancer Res., 62, pp. 910-916; Asin, S., Taylor, J.A., Trushin, S., Bren, G., Paya, C.V., (1999) J. Virol., 73, pp. 3893-3903; Bhat-Nakshatri, P., Sweeney, C.J., Nakshatri, H., (2002) Oncogene, 21, pp. 2066-2078; Bishop, A.L., Hall, A., (2000) Biochem. J., 348, pp. 241-255; Cammarano, M.S., Minden, A., (2001) J. Biol. Chem., 276, pp. 25876-25882; Cao, Y., Bonizzi, G., Seagroves, T.N., Greten, F.R., Johnson, R., Schmidt, E.V., Karin, M., (2001) Cell, 107, pp. 763-775; Chen, J.-Y., Funk, W.D., Wright, W.E., Shay, J.W., Minna, J.D., (1993) Oncogene, 8, pp. 2159-2166; Chiao, P.J., Na, R., Niu, J., Sclabas, G.M., Dong, Q.G., Curley, S.A., (2002) Cancer, 95, pp. 1696-1705; Chuang, S.-E., Yeh, P.-Y., Lu, Y.-S., Lai, G.-M., Liao, C.-M., Gao, M., Cheng, A.-L., (2002) Biochem. Pharmacol., 63, pp. 1709-1716; DiDonato, J.A., Hayakawa, M., Rothwarf, D.M., Zandi, E., Karin, M., (1997) Nature, 388, pp. 548-554; Engers, R., Zwaka, T.P., Gohr, L., Weber, A., Gerharz, C.D., Gabbert, H.E., (2000) Int. J. Cancer, 88, pp. 369-376; Fritz, G., Brachetti, C., Schmidt, M., Kaina, B., (2002) Br. J. Cancer, 87, pp. 635-644; Fritz, G., Just, I., Kaina, B., (1999) Int. J. Cancer, 81, pp. 682-687; Gildea, J.J., Seraj, M.J., Oxford, G., Harding, M.A., Hampton, G.M., Moskaluk, C.A., Frierson, H.F., Theodorescu, D., (2002) Cancer Res., 62, pp. 6418-6423; Ikeda, H., Nagashima, K., Yanase, M., Tomiya, T., Arai, M., Inoue, Y., Tejima, K., Fujiwara, K., (2003) Am. J. Physiol. Gastrointest. Liver Physiol. On-line; Ishizaki, T., Fujisawa, K., Maekawa, M., Watanabe, N., Saito, Y., Narumiya, S., (1997) FEBS Lett., 404, pp. 118-124; Kamai, T., Arai, S., Sumi, T., Tsujii, M., Honda, T., Yamanishi, T., Yoshida, K.I., (2003) BJU Int., 89, pp. 449-453; Kanai, M., Konda, Y., Nakajima, T., Izumi, Y., Takeuchi, T., Chiba, T., (2001) Gastroenterology, 121, pp. 56-67; Kaneko, K., Satoh, K., Masamune, A., Satoh, A., Shimosegawa, T., (2002) Pancreas, 24, pp. 251-257; Karin, M., Cao, Y., Greten, F.R., Li, Z.W., (2002) Nat. Rev. Cancer, 2, pp. 301-310; Kim, J.Y., Lee, S., Hwangbo, B., Lee, C.T., Kim, Y.W., Han, S.K., Shim, Y.S., Yoo, C.G., (2000) Biochem. Biophys. Res. Commun., 273, pp. 140-146; Kleer, C.G., Van Golen, K.L., Zhang, Y., Wu, Z.F., Ma, R., Merajver, S.D., (2002) Am. J. Pathol., 160, pp. 579-584; Kourlas, P.J., Strout, M.P., Becknell, B., Veronese, M.L., Croce, C.M., Theil, K.S., Krahe, R., Caligiuri, M.A., (2000) Proc. Natl. Acad. Sci. USA, 97, pp. 2145-2150; Kozma, R., Ahmed, S., Best, A., Lim, L., (1995) Mol. Cell. Biol., 15, pp. 1942-1952; Lai, J.-M., Lu, C.-Y., Yen-Yang, H.-F., Chang, Z.-F., (2001) Biochem. J., 359, pp. 227-233; Liu, C.-A., Wang, M.-J., Chi, C.-W., Wu, B., Chen, J.-Y., (2004) J. Biomed. Sci., 11, pp. 661-670; Machesky, L.M., Insall, R.H., (1998) Curr. 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PY - 2004

Y1 - 2004

N2 - Rhotekin (RTKN), the gene coding for the Rho effector, RTKN, was shown to be overexpressed in human gastric cancer (GC). In this study, we further showed that RTKN is expressed at a low level in normal cells and is overexpressed in many cancer-derived cell lines. The function of RTKN as an effector protein in Rho GTPase-mediated pathways regulating apoptosis was investigated. By transfection and expression of RTKN in cells that expressed endogenous RTKN at a low basal level, we showed that RTKN overexpression conferred cell resistance to apoptosis induced by serum deprivation or treatment with sodium butyrate, and the increased resistance correlated to the level of RTKN. Conversely, reducing RTKN expression by small interfering RNAs greatly sensitized cells to apoptosis. The RTKN-mediated antiapoptotic effect was blocked by the nuclear factor-κB (NF-κB) inhibitors, curcumin or parthenolide, but not by the phosphatidylinositol 3′-OH-kinase inhibitor, LY294002, or the MAP kinase inhibitor, PD98059. Reporter gene assays and electrophoretic mobility shift assay confirmed that RTKN overexpression led to constitutive activation of NF-κB through the phosphorylation of IκB by IKKβ. By using the RTKN truncation mutants, we showed that RTKN mediated Rho activity eliciting signaling pathway to activate NF-κB, with a concomitant induction of expression of the NF-κB antiapoptotic genes, cIAP-2, BCl-xL, A1, and A20. Consistent with these data, RTKN-expressing cells showed increased chemoresistance to 5-fluorouracil and paclitaxol, and the resistance was greatly attenuated by NF-κB inhibitor. In conclusion, overactivated Rho/ RTKN/NF-κB signaling pathway through overexpression of RTKN may play a key role in gastric tumorigenesis by conferring cells resistance to apoptosis, and this signaling pathway may serve as an important target for novel therapeutic approaches to the treatment of human GC.

AB - Rhotekin (RTKN), the gene coding for the Rho effector, RTKN, was shown to be overexpressed in human gastric cancer (GC). In this study, we further showed that RTKN is expressed at a low level in normal cells and is overexpressed in many cancer-derived cell lines. The function of RTKN as an effector protein in Rho GTPase-mediated pathways regulating apoptosis was investigated. By transfection and expression of RTKN in cells that expressed endogenous RTKN at a low basal level, we showed that RTKN overexpression conferred cell resistance to apoptosis induced by serum deprivation or treatment with sodium butyrate, and the increased resistance correlated to the level of RTKN. Conversely, reducing RTKN expression by small interfering RNAs greatly sensitized cells to apoptosis. The RTKN-mediated antiapoptotic effect was blocked by the nuclear factor-κB (NF-κB) inhibitors, curcumin or parthenolide, but not by the phosphatidylinositol 3′-OH-kinase inhibitor, LY294002, or the MAP kinase inhibitor, PD98059. Reporter gene assays and electrophoretic mobility shift assay confirmed that RTKN overexpression led to constitutive activation of NF-κB through the phosphorylation of IκB by IKKβ. By using the RTKN truncation mutants, we showed that RTKN mediated Rho activity eliciting signaling pathway to activate NF-κB, with a concomitant induction of expression of the NF-κB antiapoptotic genes, cIAP-2, BCl-xL, A1, and A20. Consistent with these data, RTKN-expressing cells showed increased chemoresistance to 5-fluorouracil and paclitaxol, and the resistance was greatly attenuated by NF-κB inhibitor. In conclusion, overactivated Rho/ RTKN/NF-κB signaling pathway through overexpression of RTKN may play a key role in gastric tumorigenesis by conferring cells resistance to apoptosis, and this signaling pathway may serve as an important target for novel therapeutic approaches to the treatment of human GC.

KW - Antiapoptosis

KW - Gastric cancer

KW - Nuclear factor-κB

KW - Rho GTPases

KW - Rhotekin

KW - 2 (2 amino 3 methoxyphenyl)chromone

KW - 2 morpholino 8 phenylchromone

KW - butyric acid

KW - curcumin

KW - fluorouracil

KW - I kappa B

KW - immunoglobulin enhancer binding protein

KW - mitogen activated protein kinase inhibitor

KW - paclitaxel

KW - parthenolide

KW - phosphatidylinositol 3 kinase inhibitor

KW - protein

KW - Rho factor

KW - Rho guanine nucleotide binding protein

KW - rhotekin

KW - small interfering RNA

KW - unclassified drug

KW - A1 gene

KW - a20 gene

KW - apoptosis

KW - article

KW - assay

KW - bcl xl gene

KW - cancer cell culture

KW - ciap 2 gene

KW - controlled study

KW - correlation analysis

KW - gel mobility shift assay

KW - gene

KW - gene expression

KW - gene function

KW - gene induction

KW - gene overexpression

KW - genetic transfection

KW - human

KW - human cell

KW - mutant

KW - nucleotide sequence

KW - priority journal

KW - protein phosphorylation

KW - regulatory mechanism

KW - reporter gene

KW - rhotekin gene

KW - serum

KW - signal transduction

KW - Amino Acid Sequence

KW - Apoptosis

KW - Base Sequence

KW - Cell Line, Tumor

KW - DNA Primers

KW - Electrophoretic Mobility Shift Assay

KW - Flow Cytometry

KW - Humans

KW - Intracellular Signaling Peptides and Proteins

KW - NF-kappa B

KW - Reverse Transcriptase Polymerase Chain Reaction

KW - rho GTP-Binding Proteins

U2 - 10.1038/sj.onc.1208106

DO - 10.1038/sj.onc.1208106

M3 - Article

VL - 23

SP - 8731

EP - 8742

JO - Oncogene

JF - Oncogene

SN - 0950-9232

IS - 54

ER -