Silencing of Lactotransferrin expression by methylation in prostate cancer progression

Syed Shaheduzzaman; Anu Vishwanath; Bungo Furusato; Jennifer Cullen; Yongmei Chen; Lionel Bañez; Martin Nau; Lakshmi Ravindranath; Kee Hong Kim; Ahmed Mohammed; Yidong Chen; Mathias Ehrich; Vasantha Srikantan; Isabell A. Sesterhenn; David G. McLeod; Maryanne Vahey; Gyorgy Petrovics; Albert Dobi; Shiv Srivastava

doi:10.4161/cbt.6.7.4327

Silencing of Lactotransferrin expression by methylation in prostate cancer progression

Syed Shaheduzzaman, Anu Vishwanath, Bungo Furusato, Jennifer Cullen, Yongmei Chen, Lionel Bañez, Martin Nau, Lakshmi Ravindranath, Kee Hong Kim, Ahmed Mohammed, Yidong Chen, Mathias Ehrich, Vasantha Srikantan, Isabell A. Sesterhenn, David G. McLeod, Maryanne Vahey, Gyorgy Petrovics, Albert Dobi^*, Shiv Srivastava

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

47 Scopus citations

Abstract

Background: Cancer cells gain selection advantages by the coordinated silencing of protective and by the activation of cell proliferation/cell survival genes. Evaluations of epithelial cell transcriptome of benign and malignant prostate glands by laser capture microdissection (LCM) identified Lactotransferrin (LTF) as the most significantly downregulated gene in prostate cancer (CaP) cells (p < 10^-6). Frequent downregulation, significant association of LTF with PSA recurrence-free survival in CaP patients and the established anti-tumorigenic effects of LTF in experimental cancer models have provided impetus to evaluate LTF expression features and mechanisms in CaP specimens. Methods: LTF mRNA expression analysis was performed in LCM derived benign and malignant prostate epithelial cells by using Affymetrix GeneChip and QRT-PCR. LTF protein expression was assessed in tissue specimens by immunohistochemistry and in serum samples from CaP patients compared to healthy male control by using ELISA. Mechanism of LTF downregulation was analyzed in 5-azadeoxycytidine treated LNCaP and LAPC4 cells using MALDI-TOF MS. Proliferation and cell cycle analysis of CaP cells by FACS flow cytrometry was assessed in LNCaP cell cultures. Results: Quantitative analysis of LTF mRNA expression in tumor cells revealed marked downregulation of LTF with significant associations to decreased PSA recurrence-free survival of CaP patients (n = 100, p ≤ 0.0322). Moreover, low levels of LTF protein expression was observed in tumor tissues as well as in sera from CaP patients (p ≤ 0.0001). LTF promoter downstream CpG island methylation was found in LNCaP and LAPC4 cells. Furthermore, replenishing of LTF by supplementing growth media with LTF protein resulted in reduced cell growth. Cell cycle analysis revealed robust increases in apoptosis in response to LTF treatment. Conclusion: This study highlights the potential for LTF in chemoprevention and to become a biologically relevant prognostic marker of CaP, suggesting that silencing of the LTF gene may be causally linked to CaP progression.

Original language	English
Pages (from-to)	1088-1095
Number of pages	8
Journal	Cancer Biology and Therapy
Volume	6
Issue number	7
DOIs	https://doi.org/10.4161/cbt.6.7.4327
State	Published - Jul 2007
Externally published	Yes

Keywords

Chemoprevention
Downregulation
LTF
Methylation
PSA doubling time

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10.4161/cbt.6.7.4327

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Shaheduzzaman, S., Vishwanath, A., Furusato, B., Cullen, J., Chen, Y., Bañez, L., Nau, M., Ravindranath, L., Kim, K. H., Mohammed, A., Chen, Y., Ehrich, M., Srikantan, V., Sesterhenn, I. A., McLeod, D. G., Vahey, M., Petrovics, G., Dobi, A., & Srivastava, S. (2007). Silencing of Lactotransferrin expression by methylation in prostate cancer progression. Cancer Biology and Therapy, 6(7), 1088-1095. https://doi.org/10.4161/cbt.6.7.4327

@article{e3b3600bd42b4ad39525297bd534ef83,

title = "Silencing of Lactotransferrin expression by methylation in prostate cancer progression",

abstract = "Background: Cancer cells gain selection advantages by the coordinated silencing of protective and by the activation of cell proliferation/cell survival genes. Evaluations of epithelial cell transcriptome of benign and malignant prostate glands by laser capture microdissection (LCM) identified Lactotransferrin (LTF) as the most significantly downregulated gene in prostate cancer (CaP) cells (p < 10-6). Frequent downregulation, significant association of LTF with PSA recurrence-free survival in CaP patients and the established anti-tumorigenic effects of LTF in experimental cancer models have provided impetus to evaluate LTF expression features and mechanisms in CaP specimens. Methods: LTF mRNA expression analysis was performed in LCM derived benign and malignant prostate epithelial cells by using Affymetrix GeneChip and QRT-PCR. LTF protein expression was assessed in tissue specimens by immunohistochemistry and in serum samples from CaP patients compared to healthy male control by using ELISA. Mechanism of LTF downregulation was analyzed in 5-azadeoxycytidine treated LNCaP and LAPC4 cells using MALDI-TOF MS. Proliferation and cell cycle analysis of CaP cells by FACS flow cytrometry was assessed in LNCaP cell cultures. Results: Quantitative analysis of LTF mRNA expression in tumor cells revealed marked downregulation of LTF with significant associations to decreased PSA recurrence-free survival of CaP patients (n = 100, p ≤ 0.0322). Moreover, low levels of LTF protein expression was observed in tumor tissues as well as in sera from CaP patients (p ≤ 0.0001). LTF promoter downstream CpG island methylation was found in LNCaP and LAPC4 cells. Furthermore, replenishing of LTF by supplementing growth media with LTF protein resulted in reduced cell growth. Cell cycle analysis revealed robust increases in apoptosis in response to LTF treatment. Conclusion: This study highlights the potential for LTF in chemoprevention and to become a biologically relevant prognostic marker of CaP, suggesting that silencing of the LTF gene may be causally linked to CaP progression.",

keywords = "Chemoprevention, Downregulation, LTF, Methylation, PSA doubling time",

author = "Syed Shaheduzzaman and Anu Vishwanath and Bungo Furusato and Jennifer Cullen and Yongmei Chen and Lionel Ba{\~n}ez and Martin Nau and Lakshmi Ravindranath and Kim, {Kee Hong} and Ahmed Mohammed and Yidong Chen and Mathias Ehrich and Vasantha Srikantan and Sesterhenn, {Isabell A.} and McLeod, {David G.} and Maryanne Vahey and Gyorgy Petrovics and Albert Dobi and Shiv Srivastava",

note = "Funding Information: an ongoing grant from the US Army Medical Research and Material Command, and by NIH Grant RO1 DK065977. Funding Information: Financial disclosure. This work was funded by the CPDR through",

year = "2007",

month = jul,

doi = "10.4161/cbt.6.7.4327",

language = "English",

volume = "6",

pages = "1088--1095",

journal = "Cancer Biology and Therapy",

issn = "1538-4047",

number = "7",

}

Shaheduzzaman, S, Vishwanath, A, Furusato, B, Cullen, J, Chen, Y, Bañez, L, Nau, M, Ravindranath, L, Kim, KH, Mohammed, A, Chen, Y, Ehrich, M, Srikantan, V, Sesterhenn, IA, McLeod, DG, Vahey, M, Petrovics, G , Dobi, A & Srivastava, S 2007, 'Silencing of Lactotransferrin expression by methylation in prostate cancer progression', Cancer Biology and Therapy, vol. 6, no. 7, pp. 1088-1095. https://doi.org/10.4161/cbt.6.7.4327

TY - JOUR

T1 - Silencing of Lactotransferrin expression by methylation in prostate cancer progression

AU - Shaheduzzaman, Syed

AU - Vishwanath, Anu

AU - Furusato, Bungo

AU - Cullen, Jennifer

AU - Chen, Yongmei

AU - Bañez, Lionel

AU - Nau, Martin

AU - Ravindranath, Lakshmi

AU - Kim, Kee Hong

AU - Mohammed, Ahmed

AU - Chen, Yidong

AU - Ehrich, Mathias

AU - Srikantan, Vasantha

AU - Sesterhenn, Isabell A.

AU - McLeod, David G.

AU - Vahey, Maryanne

AU - Petrovics, Gyorgy

AU - Dobi, Albert

AU - Srivastava, Shiv

N1 - Funding Information: an ongoing grant from the US Army Medical Research and Material Command, and by NIH Grant RO1 DK065977. Funding Information: Financial disclosure. This work was funded by the CPDR through

PY - 2007/7

Y1 - 2007/7

N2 - Background: Cancer cells gain selection advantages by the coordinated silencing of protective and by the activation of cell proliferation/cell survival genes. Evaluations of epithelial cell transcriptome of benign and malignant prostate glands by laser capture microdissection (LCM) identified Lactotransferrin (LTF) as the most significantly downregulated gene in prostate cancer (CaP) cells (p < 10-6). Frequent downregulation, significant association of LTF with PSA recurrence-free survival in CaP patients and the established anti-tumorigenic effects of LTF in experimental cancer models have provided impetus to evaluate LTF expression features and mechanisms in CaP specimens. Methods: LTF mRNA expression analysis was performed in LCM derived benign and malignant prostate epithelial cells by using Affymetrix GeneChip and QRT-PCR. LTF protein expression was assessed in tissue specimens by immunohistochemistry and in serum samples from CaP patients compared to healthy male control by using ELISA. Mechanism of LTF downregulation was analyzed in 5-azadeoxycytidine treated LNCaP and LAPC4 cells using MALDI-TOF MS. Proliferation and cell cycle analysis of CaP cells by FACS flow cytrometry was assessed in LNCaP cell cultures. Results: Quantitative analysis of LTF mRNA expression in tumor cells revealed marked downregulation of LTF with significant associations to decreased PSA recurrence-free survival of CaP patients (n = 100, p ≤ 0.0322). Moreover, low levels of LTF protein expression was observed in tumor tissues as well as in sera from CaP patients (p ≤ 0.0001). LTF promoter downstream CpG island methylation was found in LNCaP and LAPC4 cells. Furthermore, replenishing of LTF by supplementing growth media with LTF protein resulted in reduced cell growth. Cell cycle analysis revealed robust increases in apoptosis in response to LTF treatment. Conclusion: This study highlights the potential for LTF in chemoprevention and to become a biologically relevant prognostic marker of CaP, suggesting that silencing of the LTF gene may be causally linked to CaP progression.

AB - Background: Cancer cells gain selection advantages by the coordinated silencing of protective and by the activation of cell proliferation/cell survival genes. Evaluations of epithelial cell transcriptome of benign and malignant prostate glands by laser capture microdissection (LCM) identified Lactotransferrin (LTF) as the most significantly downregulated gene in prostate cancer (CaP) cells (p < 10-6). Frequent downregulation, significant association of LTF with PSA recurrence-free survival in CaP patients and the established anti-tumorigenic effects of LTF in experimental cancer models have provided impetus to evaluate LTF expression features and mechanisms in CaP specimens. Methods: LTF mRNA expression analysis was performed in LCM derived benign and malignant prostate epithelial cells by using Affymetrix GeneChip and QRT-PCR. LTF protein expression was assessed in tissue specimens by immunohistochemistry and in serum samples from CaP patients compared to healthy male control by using ELISA. Mechanism of LTF downregulation was analyzed in 5-azadeoxycytidine treated LNCaP and LAPC4 cells using MALDI-TOF MS. Proliferation and cell cycle analysis of CaP cells by FACS flow cytrometry was assessed in LNCaP cell cultures. Results: Quantitative analysis of LTF mRNA expression in tumor cells revealed marked downregulation of LTF with significant associations to decreased PSA recurrence-free survival of CaP patients (n = 100, p ≤ 0.0322). Moreover, low levels of LTF protein expression was observed in tumor tissues as well as in sera from CaP patients (p ≤ 0.0001). LTF promoter downstream CpG island methylation was found in LNCaP and LAPC4 cells. Furthermore, replenishing of LTF by supplementing growth media with LTF protein resulted in reduced cell growth. Cell cycle analysis revealed robust increases in apoptosis in response to LTF treatment. Conclusion: This study highlights the potential for LTF in chemoprevention and to become a biologically relevant prognostic marker of CaP, suggesting that silencing of the LTF gene may be causally linked to CaP progression.

KW - Chemoprevention

KW - Downregulation

KW - LTF

KW - Methylation

KW - PSA doubling time

UR - http://www.scopus.com/inward/record.url?scp=42449093747&partnerID=8YFLogxK

U2 - 10.4161/cbt.6.7.4327

DO - 10.4161/cbt.6.7.4327

M3 - Article

C2 - 17568188

AN - SCOPUS:42449093747

SN - 1538-4047

VL - 6

SP - 1088

EP - 1095

JO - Cancer Biology and Therapy

JF - Cancer Biology and Therapy

IS - 7

ER -

Silencing of Lactotransferrin expression by methylation in prostate cancer progression

Abstract

Keywords

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