Original article
Low expression of TMEM67 is a critical predictor of poor prognosis in human urothelial carcinoma of the bladder

https://doi.org/10.1016/j.urolonc.2016.10.014Get rights and content

Highlights

  • TMEM67 expression is low in UCB tissues.

  • TMEM67 low expression predicted poor prognosis of patients with UCB.

Abstract

Objectives

The aim of the study was to evaluate the expression of TMEM67 in urothelial carcinoma of the bladder (UCB) tissues and to determine the potential relevance between the expression of TMEM67 and prognosis of UCB.

Material and methods

In this study, the expression of TMEM67 mRNA was performed by quantitative real-time PCR in 80 UCB and 54 noncancerous tissues. The expression of TMEM67 protein was identified by immunohistochemistry and western blotting. Chi-square test was conducted to verify the relevance between the expression of TMEM67 and clinical parameters. Kaplan-Meier survival analysis was demonstrated between high or low expression level of TMEM67 mRNA and recurrence-free survival probability. Cox regression analysis was conducted to evaluate the relevance between the expression of TMEM67 and the prognosis in UCB.

Results

Low expression of TMEM67 mRNA and protein was detected in most of UCB tissues using quantitative real-time polymerase chain reaction and western blotting, compared with noncancerous tissues. Low expressions of TMEM67 were associated with TNM stage, grade, and lymph node metastasis (P<0.05). Kaplan-Meier analysis showed that the low expression of TMEM67 mRNA had significantly shorter recurrence-free survival probability (P = 0.018). Cox regression analysis confirmed that low expression of TMEM67 mRNA predicted poor prognosis of patients with UCB (HR = 2.950, P = 0.029, 95% CI: 1.1167.796).

Conclusions

TMEM67 expression is low in UCB tissues, and the TMEM67 low expression predicted poor prognosis of patients with UCB.

Introduction

Urothelial carcinoma of the bladder (UCB) is the most common urothelial malignancies which accounts for 3.3% of newly diagnosed cancer cases and 2.1% of cancer deaths in the world [1]. Overall, 70% of bladder tumors present as noninvasive urothelial carcinoma, and the remainder present as muscle-invasive disease [2]. So, an accurate biomarker or prognosis factor is essential for efficient management of UCB.

TMEM67, also named Meckelin, is a transmembrane protein encoded by TMEM67/MKS3, localizing to the primary cilium and the plasma membrane [3], [4], [5]. Genotype-phenotype analyses indicated that recessive mutations at the TMEM67 locus were associated with dysfunction of primary cilia [4], [5], [6]. Primary cilia are ubiquitously present in cell surface organelles with essential functions in cellular proliferation, differentiation and development. Emerging evidences suggest that in many cancers [7], [8], [9], [10], primary cilia are markedly decreased or absent. To date, however, abnormalities in TMEM67 and their influence to UCB have not been declared.

In this study, we explored the expression status of TMEM67 in UCB and evaluated its possible biogenic role as a prognostic biomarker for patients with UCB.

Section snippets

Patients and tumor specimens

A total of 134 tissues (54 UCB and noncancerous tissues by radical cystectomy, 26 UCB tissues by TURBT) were obtained from patients with UCB, who were diagnosed by pathological examination at the Second Hospital of Tianjin Medical University from January 2005 to October 2013, and 54 noncancerous tissues were collected outside 3 cm adjacent to carcinoma tissues as control at the same time. No patient was preoperatively treated with either radiotherapy or chemotherapy and postoperatively treated

Low expression of TMEM67 in UCB tissues

To evaluate the biogenic role of TMEM67 in UCB, we examined the expression of TMEM67 in noncancerous tissues and cancer tissues by qRT-PCR, IHC, and Western blot analysis. The results showed that the expression of TMEM67 mRNA was significantly lower in UCB tissues compared to those in noncancerous tissues (Fig. 1A, P = 0.0161). As shown in Fig. 1B, the TMEM67 protein was differentially low expressed in 11 UCB samples compared to corresponding adjacent noncancerous tissues. IHC staining showed

Discussion

UCB is genetically heterogeneous [11], with frequent alterations in genes regulating chromatin state, cell cycle, and receptor kinase signaling [12], [13], [14], [15] and it is a cancer with high recurrent rate and the 5-year overall survival is still unsatisfactory for advanced disease in spite of the introduction of adjuvant chemotherapy [16]. Current patient prognosis is mainly based on TNM. The high recurrence rate requires close surveillance with cystoscopy that is an invasive test, so an

Conclusions

This study data showed that TMEM67 was significantly low expressed in UCB tissues. Low expression of TMEM67 in UCB is strongly relevance with tumor stage, grade and lymph node metastasis. TMEM67 was confirmed as a prognostic biomarker in UCB by Kaplan-Meier survival and Cox regression analysis. In combination with other biomarkers of UCB, TMEM67 might be valuable for providing potential targets for new therapeutic approaches for patients with UCB.

Acknowledgment

This work was supported by National Natural Science Foundation of China (No. 81400686), Tianjin Natural Science Foundation (No. 13JCQNJC11500).

References (34)

  • J.A. Bishop et al.

    Ciliated HPV-related carcinoma: a well-differentiated form of head and neck carcinoma that can be mistaken for a benign cyst

    Am J Surg Pathol

    (2015)
  • U.E. Lang et al.

    Loss of primary cilia correlates with cytologic severity in dysplastic melanocytic nevi

    J Cutan Pathol

    (2016)
  • D. Gate et al.

    Characterization of cancer stem cells and primary cilia in medulloblastoma

    Cns Neurol Disord Drug Targets

    (2015)
  • C.E. de Andrea et al.

    Cell cycle deregulation and mosaic loss of Ext1 drive peripheral chondrosarcomagenesis in the mouse and reveal an intrinsic cilia deficiency

    J Pathol

    (2015)
  • S.L. Park et al.

    EPO gene expression induces the proliferation, migration and invasion of bladder cancer cells through the p21WAF1mediated ERK1/2/NF-kappaB /MMP-9 pathway

    Oncol Rep

    (2014)
  • A. Greife et al.

    Canonical Notch signalling is inactive in urothelial carcinoma

    BMC Cancer

    (2014)
  • S.J. Lee et al.

    Aqueous extract of Magnolia officinalis mediates proliferative capacity, p21WAF1 expression and TNF-α-induced NF-κB activity in human urinary bladder cancer 5637 cells;involvement of p38 MAP kinase

    Oncol Rep

    (2007)
  • View full text