PBK/TOPK in the differential diagnosis of cholangiocarcinoma from hepatocellular carcinoma and its involvement in prognosis of human cholangiocarcinoma

doi:10.1016/j.humpath.2009.05.016

Human Pathology

Volume 41, Issue 3, March 2010, Pages 415-424

https://doi.org/10.1016/j.humpath.2009.05.016 Get rights and content

Summary

The increased expression of PDZ binding kinase/lymphokine-activated killer T-cell–originated protein kinase (PBK/TOPK) is associated with some human malignant tumors. In this study, we analyzed PBK/TOPK expression in hepatic primary tumor and explored its role in cholangiocarcinoma biology. Seventy-four cholangiocarcinomas, 33 hepatocellular carcinomas, and 10 normal liver tissues were prepared from paraffin-embedded specimens. PBK/TOPK protein was assessed by immunohistochemical staining, and the survival time was analyzed with the Kaplan-Meier method. The protein, mRNA of PBK/TOPK, and cell cycle of cholangiocarcinoma cell line after PBK/TOPK suppression with small interfere RNA were studied by Western blot, semiquantitative reverse transcriptase–polymerase chain reaction, and flow cytometry, respectively. PBK/TOPK was usually expressed in normal bile duct epithelial cells and much more frequently expressed in cholangiocarcinoma (68/74) but never expressed in hepatocytes and hepatocellular carcinomas (0/33). PBK/TOPK down-regulation was related to the poor prognosis of patients with cholangiocarcinoma (P = .013). Epidermal growth factor can enhance PBK/TOPK expression in cholangiocarcinoma QBC 939 cells, but suppression of PBK/TOPK in the cells did not affect their proliferation. PBK/TOPK protein could serve as a useful indicator for histopathologic differentiation between cholangiocarcinoma and hepatocellular carcinomas and the low expression of PBK/TOPK is predicative of poor survival in cholangiocarcinoma patients.

Introduction

PDZ binding kinase/lymphokine-activated killer T-cell–originated protein kinase (PBK/TOPK) is a 322 amino acid mitotic protein kinase and a member of the MEK protein family. It is reported that PBK/TOPK can phosphorylate p38 and is suggested to be a novel MEK3/6-related mitogen-activated protein kinase [1], [2]. PBK/TOPK is hardly detected in normal tissue except the testis, but it is overexpressed in hematopoietic neoplasms, including Burkitt's lymphoma, acute lymphoblastic leukemia, multiple myeloma and promyelocytic leukemia, and some cancers such as breast carcinoma and colorectal cancer [3], [4]. Although PBK/TOPK is closely related to the above mentioned malignancies, its mechanism is still unclear. It is reported that PBK/TOPK is involved in the cytokinetic function and in DNA damage and repair [5], [6]. In sum, previous studies have discovered that PBK/TOPK, which is identified as a mitogen-activated protein kinase kinase, plays an important role in some malignant tumors and is suggested to be a molecular target for malignancy.

Cholangiocarcinoma (CC), a primary liver tumor that arises from biliary epithelial cells, increases in incidence and has poor prognosis. Unlike hepatocellular carcinoma (HCC), no predisposing factors or high-risk populations have been demonstrated for cholangiocarcinoma [7]. Recent data show that the incidence and mortality rates of intrahepatic CC (ICC) increase in several areas around the world [8], [9]. Timely diagnosis of cholangiocarcinoma is essential because surgical resection in early disease remains the only cure. Lack of a sensitive and specific early diagnostic marker as well as alternative treatment is the main reason why patients have limited survival time. The use of diverse approaches, through which are analyzed the physiological or pathological complement of proteins in cells, tissues, or biological fluids, has received substantial interest in biomarker discovery for cholangiocarcinoma [10].

In recent studies, PBK/TOPK mRNA has been detected at low level in liver tissue [2], but PBK/TOPK protein and its function have not been revealed in the liver. In our previous work, PBK/TOPK expression was detected in 19 types of both normal and tumor tissues by using tissue chip assay and immunohistochemical staining. We found that PBK/TOPK was only expressed in normal bile duct cell and cholangiocarcinoma, so we used many surgical specimens of cholangiocarcinoma and HCC to analyze PBK/TOPK expression and survival time of the patients with cholangiocarcinoma. We found, for the first time, that PBK/TOPK was only expressed in cholangiocarcinoma tissues and its down-regulation was significantly related to prognosis of the patients, which suggested that PBK/TOPK could be used as an indicator for diagnosis and prognosis of cholangiocarcinoma. In addition, a preliminary study on the function of PBK/TOPK in cholangiocarcinoma cell line has been performed.

Section snippets

Patients and clinical samples

One hundred seven liver tumors and 10 normal liver tissues (100 surgical specimens and 17 needle biopsies) during 2003 to 2007 in Xijing Hospital were included in our retrospective study. The specimens were composed of 74 CCs (including 17 needle biopsies), 33 HCCs, and 10 normal liver tissues obtained from the donor livers. All the tissues were retrieved from the consecutive patients of the pathology archives in Xijing Hospital. These specimens came from 82 male patients and 35 female patients

Clinical data and histopathology

Human cholangiocarcinoma specimens were composed of 51 cases of ICC (34 surgical specimens and 17 needle biopsies) and 23 cases of hilar CC. All the HCC cases were surgical specimens and the normal liver tissues were obtained from the donor livers. Histological grading of cholangiocarcinoma was assessed based on the World Health Organization Classification of Tumors [16]. In 57 cases of the cholangiocarcinoma specimens, 39 cases were well or intermediately differentiated (Fig. 1A and B), and 18

Discussion

Cholangiocarcinoma is the second most frequent primary malignant epithelial liver tumor [20]. In addition, intrahepatic cholangiocarcinoma is increasing worldwide [9], [21]. The diagnosis, poor prognosis, evaluation and management of these tumors are still challenges [22], [23] and molecular mechanisms underlying the development, growth and metastasis in biliary tract cancer are still unclear. The worldwide increase in incidence, mortality and poor prognosis of cholangiocarcinoma impel us to

Acknowledgments

We are very grateful to Professor Yumei Zhou and Donglei Jiang (Department of Foreign Languages, Fourth Military Medical University) for their help in English writing.

References (34)

AbeY. et al.
Cloning and expression of a novel MAPKK-like protein kinase, lymphokine-activated killer T-cell-originated protein kinase, specifically expressed in the testis and activated lymphoid cells
J Biol Chem
(2000)
GaudetS. et al.
Characterization of PDZ-binding kinase, a mitotic kinase
Proc Natl Acad Sci U S A
(2000)
ParkJ.H. et al.
PDZ-binding kinase/T-LAK cell-originated protein kinase, a putative cancer/testis antigen with an oncogenic activity in breast cancer
Cancer Res
(2006)
ZhuF. et al.
Bidirectional signals transduced by TOPK-ERK interaction increase tumorigenesis of HCT116 colorectal cancer cells
Gastroenterology
(2007)
AyllonV. et al.
PBK/TOPK promotes tumour cell proliferation through p38 MAPK activity and regulation of the DNA damage response
Oncogene
(2007)
AbeY. et al.
A mitotic kinase TOPK enhances Cdk1/cyclin B1-dependent phosphorylation of PRC1 and promotes cytokinesis
J Mol Biol
(2007)
MetairieS. et al.
Intra-hepatic cholangiocarcinoma
J Chir (Paris)
(2004)
MartinR. et al.
Intrahepatic cholangiocarcinoma: Current management
Minerva Chir
(2003)
PatelT.
Increasing incidence and mortality of primary intrahepatic cholangiocarcinoma in the United States
Hepatology
(2001)
BonneyG.K. et al.
Circulating markers of biliary malignancy: opportunities in proteomics?
Lancet Oncol
(2008)

WangJ.H. et al.

Detection of human parvovirus B19 in papillary thyroid carcinoma

Br J Cancer

(2008)

BrownD.C. et al.

Ki67 protein: the immaculate deception?

Histopathology

(2002)

AngelopoulouR. et al.

Evaluation of immunohistochemical markers of germ cells' proliferation in the developing rat testis: a comparative study

Tissue Cell

(2008)

ShiZ. et al.

Antisense RhoC gene suppresses proliferation and invasion capacity of human QBC939 cholangiocarcinoma cells

Hepatobiliary Pancreat Dis Int

(2007)

LiY. et al.

Detection of parvovirus B19 nucleic acids and expression of viral VP1/VP2 antigen in human colon carcinoma

Am J Gastroenterol

(2007)

NakanumaY. et al.

Intrahepatic cholangiocarcinoma

HirohashiS. et al.

Hepatocellular carcinoma

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Cell growth inhibition by 3-deoxysappanchalcone is mediated by directly targeting the TOPK signaling pathway in colon cancer
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Colorectal cancer is one of the most common causes of cancer death worldwide. Unfortunately, chemotherapies are limited due to many complications and development of resistance and recurrence. The T-lymphokine-activated killer cell-originated protein kinase (TOPK) is highly expressed and activated in colon cancer, and plays an important role in inflammation, proliferation, and survival of cancer cells. Therefore, suppressing TOPK activity and its downstream signaling cascades is considered to be a rational therapeutic/preventive strategy against colon cancers.
3-Deoxysappanchalcone (3-DSC), a component of Caesalpinia sappan L., is a natural oriental medicine. In this study, we investigated the effects of 3-DSC on colon cancer cell growth and elucidated its underlying molecular mechanism of targeting TOPK.
To evaluate the effects of 3-DSC against colon cancer, we performed cell proliferation assays, propidium iodide- and annexin V-staining analyses and Western blotting. Targeting TOPK by 3-DSC was identified by a kinase-binding assay and computational docking models.
3-DSC inhibited the kinase activity of TOPK, but not mitogen-activated protein kinase (MEK). The direct binding of 3-DSC with TOPK was explored using a computational docking model and binding assay in vitro and ex vivo. 3-DSC inhibited colon cancer cell proliferation and anchorage-independent cell growth, and induced G2/M cell cycle arrest and apoptosis. Treatment of colon cancer cells with 3-DSC induced expression of protein that are involved in cell cycle (cyclin B1) and apoptosis (cleaved-PARP, cleaved-caspase-3, and cleaved-caspase-7), and suppressed protein expressions of extracellular signal-regulated kinase (ERK)-1/2, ribosomal S6 kinase (RSK), and c-Jun, which are regulated by the upstream kinase, TOPK.
3-DSC suppresses colon cancer cell growth by directly targeting the TOPK- mediated signaling pathway.
Novel selective TOPK inhibitor SKLB-C05 inhibits colorectal carcinoma growth and metastasis
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Citation Excerpt :
In this context, TOPK serves as an upstream activator of MAPK and mitosis relevant kinase, and plays a pivotal role in cancer-advancing activities such as tumor proliferation, apoptosis, inflammation, and metastasis [10,12–15]. TOPK was reported aberrantly expressed in cancers such as CRC [16], breast [17], prostate [18], lung [19], and cholangiocarcinoma [20], etc. CRC is one of the most common cancer types worldwide.
The mitogen-activated protein kinase (MAPK) signaling pathway member T-LAK cell–originated protein kinase/PDZ-binding kinase (TOPK/PBK) is closely involved in tumorigenesis and progression. Its overexpression in colorectal carcinoma (CRC) exacerbates tumor malignancy, promotes metastasis and results in dismal prognosis. Therefore, targeting TOPK is a promising approach for CRC therapy. Here, we report the development of a TOPK selective inhibitor SKLB-C05, with subnanomolar inhibitory potency. In vitro, SKLB-C05 exhibited excellent cytotoxicity and anti-migration and invasion activity on TOPK high-expressing CRC cells and induced cell apoptosis. These activities could attribute to its inhibition of TOPK downstream signaling including extracellular signal-regulated kinase 1/2 (ERK1/2), p38, and c-Jun N-terminal kinase 1, 2, and 3 (JNK1/2/3), as well as downregulation of FAK/Src- MMP signaling. Furthermore, SKLB-C05 disrupted cell mitosis and blocked CRC cell cycle. In vivo, oral administration of SKLB-C05 at concentrations of 20 and 10 mg kg⁻¹·day⁻¹ dramatically attenuated CRC tumor xenograft growth and completely suppressed hepatic metastasis of HCT116 cells, respectively. Thus, these findings suggest that SKLB-C05 is a specific TOPK inhibitor with potent anti-CRC oncogenic activity in vitro and in vivo.
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This network includes genes such as cluster of differentiation 80 (CD80), which regulates T-cell activation by binding to cytotoxic T-lymphocyte antigen-4 (CTLA4) (Suppl Fig. 1). Amongst the transcripts deregulated by an average >2-fold (Suppl Table 1, Fig. 1B), there is PDZ-binding kinase, a kinase produced by lymphokine-activated killer T-cells, which was previously found to be a CCA-specific transcript associated with prognosis [19]. It is recognised that deregulation of the immune system may be a cause of cancer progression by creating a favourable microenvironment for cancer growth and escape.
Although biliary tract cancers (BTCs) are known to have an inflammatory component, a detailed characterisation of immune-related transcripts has never been performed. In these studies, nCounter PanCancer Immune Profiling Panel was used to assess the expression of 770 immune-related transcripts in the tumour tissues (TTs) and matched adjacent tissues (ATs) of resected BTCs. Cox regression analysis and Kaplan–Meier methods were used to correlate findings with relapse-free survival (RFS). The first analysis in the TT and AT of an exploratory set (n = 22) showed deregulation of 39 transcripts associated with T-cell activation. Risk of recurrence was associated with a greater number of genes deregulated in AT in comparison to TT. Analysis in the whole set (n = 53) showed a correlation between AT cytotoxic T-lymphocyte antigen-4 (CTLA4) expression and RFS, which maintained statistical significance at multivariate analysis. CTLA4 expression correlated with forkhead box P3 (FOXP3) expression, suggesting enrichment in T regulatory cells. CTLA4 is known to act by binding to the cluster of differentiation 80 (CD80). No association was seen between AT CD80 expression and RFS. However, CD80 expression differentiated prognosis in patients who received adjuvant chemotherapy. We showed that the immunomodulatory transcriptome is deregulated in resected BTCs. Our study includes a small number of patients and does not enable to draw definitive conclusions; however, it provides useful insights into potential transcripts that may deserve further investigation in larger cohorts of patients.
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Original contributionPBK/TOPK in the differential diagnosis of cholangiocarcinoma from hepatocellular carcinoma and its involvement in prognosis of human cholangiocarcinoma

Summary

Introduction

Section snippets

Patients and clinical samples

Clinical data and histopathology

Discussion

Acknowledgments

Cloning and expression of a novel MAPKK-like protein kinase, lymphokine-activated killer T-cell-originated protein kinase, specifically expressed in the testis and activated lymphoid cells

J Biol Chem

Characterization of PDZ-binding kinase, a mitotic kinase

Proc Natl Acad Sci U S A

PDZ-binding kinase/T-LAK cell-originated protein kinase, a putative cancer/testis antigen with an oncogenic activity in breast cancer

Cancer Res

Bidirectional signals transduced by TOPK-ERK interaction increase tumorigenesis of HCT116 colorectal cancer cells

Gastroenterology

PBK/TOPK promotes tumour cell proliferation through p38 MAPK activity and regulation of the DNA damage response

Oncogene

A mitotic kinase TOPK enhances Cdk1/cyclin B1-dependent phosphorylation of PRC1 and promotes cytokinesis

J Mol Biol

Intra-hepatic cholangiocarcinoma

J Chir (Paris)

Intrahepatic cholangiocarcinoma: Current management

Minerva Chir

Increasing incidence and mortality of primary intrahepatic cholangiocarcinoma in the United States

Hepatology

Circulating markers of biliary malignancy: opportunities in proteomics?

Lancet Oncol