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Quantitation of rare circulating tumor cells by folate receptor α ligand-targeted PCR in bladder transitional cell carcinoma and its potential diagnostic significance

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Tumor Biology

Abstract

Numerous attempts for detection of circulating tumor cells (CTC) have been made to develop reliable assays for early diagnosis of cancers. In this study, we validated the application of folate receptor α (FRα) as the tumor marker to detect CTC through tumor-specific ligand PCR (LT-PCR) and assessed its utility for diagnosis of bladder transitional cell carcinoma (TCC). Immunohistochemistry for FRα was performed on ten bladder TCC tissues. Enzyme-linked immunosorbent assay (ELISA) for FRα was performed on both urine and serum specimens from bladder TCC patients (n = 64 and n = 20, respectively) and healthy volunteers (n = 20 and n = 23, respectively). Western blot analysis and qRT-PCR were performed to confirm the expression of FRα in bladder TCC cells. CTC values in 3-mL peripheral blood were measured in 57 bladder TCC patients, 48 healthy volunteers, and 15 subjects with benign urologic pathologies by the folate receptor α ligand-targeted PCR. We found that FRα protein was overexpressed in both bladder TCC cells and tissues. The levels of FRα mRNA were also much higher in bladder cancer cell lines 5637 and SW780 than those of leukocyte. Values of FRα were higher in both serum and urine specimens of bladder TCC patients than those of control. CTC values were also higher in 3-mL peripheral blood of bladder TCC patients than those of control (median 26.5 Cu/3 mL vs 14.0 Cu/3 mL). Area under the receiver operating characteristic (ROC) curve for bladder TCC detection was 0.819, 95 % CI (0.738–0.883). At the cutoff value of 15.43 Cu/3 mL, the sensitivity and the specificity for detecting bladder cancer are 82.14 and 61.9 %, respectively. We concluded that quantitation of CTCs through FRα ligand-PCR could be a promising method for noninvasive diagnosis of bladder TCC.

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Acknowledgments

This work was supported by grants from the National Nature Science Foundation of China (NSFC 81372507 and 81101584), National Basic Research Program of China (973 Program) (Grant no. 338 2014CB745200), Shenzhen Peacock Foundation (KQC201106020032A), and Shenzhen Basic Research Key Project (JCYJ20120614455816670). International S&T Cooperation Program of China (2014DFA31050).

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Authors and Affiliations

  1. Department of Urological Surgery, Shenzhen Second People’s Hospital, The First Affiliated Hospital of Shenzhen University, Shenzhen, China

    Fuming Qi, Yuchen Liu, Xiangjun Zou & Zhiming Cai

  2. Guangdong and Shenzhen Key Laboratory of Male Reproductive Medicine and Genetics, Institute of Urology, Peking University Shenzhen Hospital, Shenzhen PKU-HKUST Medical Center, Shenzhen, China

    Fuming Qi, Yuchen Liu, Jiaqiang Li, Ye Xia, Xuliang Wang, Cailing Li & Yaoting Gui

  3. Shantou University Medical College, Shantou, China

    Fuming Qi

  4. Shenzhen Key Laboratory of Genitourinary Tumor, Shenzhen Second People’s Hospital, The First Affiliated Hospital of Shenzhen University, Shenzhen, 518035, China

    Yuchen Liu, Rongchang Zhao, Xiangjun Zou, Lei Zhang, Jiaqiang Li, Yongqiang Wang, Feiyang Li, Xiaowen Zou, Ye Xia, Xuliang Wang, Li Xing & Zhiming Cai

  5. Department of Urology, Sun Yat-Sen University Cancer Center, Guangzhou, China

    Fangjian Zhou

  6. Department of Urology, Zhujiang Hospital of Southern Medical University, Guangzhou, China

    Lei Zhang, Li Xing & Chunxiao Liu

  7. Department of Research and Development, GenoSaber Biotech Co., Ltd., Shanghai, China

    Rongchang Zhao

  8. Shenzhen Tumor Immuno-gene Therapy Clininical Application Engineering Lab, Biobank of Shenzhen Second People’s Hospital, The First Affiliated Hospital of Shenzhen University, 802 Room, Yinhua Building, 3002 West Sungang Road, Futian District, Shenzhen, 518035, China

    Jingxiao Lu, Junlong Tang & Xiaojuan Sun

Authors
  1. Fuming Qi
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  2. Yuchen Liu
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  3. Rongchang Zhao
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  5. Lei Zhang
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  6. Jiaqiang Li
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  7. Yongqiang Wang
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  8. Feiyang Li
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  9. Xiaowen Zou
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  10. Ye Xia
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  11. Xuliang Wang
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  12. Li Xing
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  13. Cailing Li
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  14. Jingxiao Lu
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  15. Junlong Tang
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  16. Fangjian Zhou
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  17. Chunxiao Liu
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  18. Yaoting Gui
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  19. Zhiming Cai
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  20. Xiaojuan Sun
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Corresponding authors

Correspondence to Zhiming Cai or Xiaojuan Sun.

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Fuming Qi and Yuchen Liu contributed equally to this work.

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Qi, F., Liu, Y., Zhao, R. et al. Quantitation of rare circulating tumor cells by folate receptor α ligand-targeted PCR in bladder transitional cell carcinoma and its potential diagnostic significance. Tumor Biol. 35, 7217–7223 (2014). https://doi.org/10.1007/s13277-014-1894-0

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  • DOI: https://doi.org/10.1007/s13277-014-1894-0

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