Share This Article:

Usefulness of Immuno-Magnetic Beads Conjugated with Anti-EpCAM Antibody for Detecting Endometrial Cancer Cells

Abstract Full-Text HTML Download Download as PDF (Size:948KB) PP. 1273-1282
DOI: 10.4236/jct.2013.48150    3,954 Downloads   5,720 Views   Citations

ABSTRACT

A simple and non-invasive method for detecting endometrial cancer in women with abnormal uterine bleeding is required. For this purpose, we prepared immuno-magnetic beads conjugated with anti-human EpCAM rat monoclonal antibody (mAb) for isolating exfoliated endometrial cells including endometrial cancer cells in vaginal discharge. The affinities of the anti-human EpCAM rat mAbs were analyzed by flow cytometry and immunocytochemistry and then magnetic beads were conjugated with the mAbs. The rate of retrieval of endometrial cells using the immuno-magnetic beads was calculated. Endometrial cells were isolated using the immuno-magnetic beads from the vaginal discharges of 22 patients with endometrial cancer and 16 non-malignant controls. The isolated cells were stained using endometrial cancer specific-mAbs and analyzed by flow cytometry and imaging cytometry. The immuno-magnetic beads conjugated with high-affinity mAb (clone 1456) appeared to have very low auto-fluorescence. Sufficient enrichment of Ep-CAMpositive cells using immuno-magnetic beads was observed in both simulation and clinical samples. The overall sensitivities of flow cytometry and imaging cytometry to detect endometrial cancer cells were 72.7% and 45.5%, respectively. Meanwhile, the overall specificities of flow cytometry and imaging cytometry for healthy controls were 75.0% and 81.3%, respectively. Our immuno-magnetic beads have very low auto-fluorescence, so they could be useful for fluorescent analysis, such as fluorescent immunochemical staining. In the future, these novel immuno-magnetic beads could be used for cytological study.

Conflicts of Interest

The authors declare no conflicts of interest.

Cite this paper

Y. Koga, S. Katayose, N. Onoda, T. Kasamatsu, T. Kato, S. Ikeda, M. Ishikawa, K. Ishitani, Y. Hirai, H. Matsui and Y. Matsumura, "Usefulness of Immuno-Magnetic Beads Conjugated with Anti-EpCAM Antibody for Detecting Endometrial Cancer Cells," Journal of Cancer Therapy, Vol. 4 No. 8, 2013, pp. 1273-1282. doi: 10.4236/jct.2013.48150.

References

[1] F. Amant, P. Moerman, P. Neven, D. Timmerman, E. Van Limbergen and I. Vergote, “Endometrial Cancer,” Lancet, Vol. 366, No. 9484, 2005, pp. 491-505. http://dx.doi.org/10.1016/S0140-6736(05)67063-8
[2] R. Siegel, D. Naishadham and A. Jemal, “Cancer Statistics, 2013,” CA: A Cancer Journal for Clinicians, Vol. 63, No. 1, 2013, pp. 11-30. http://dx.doi.org/10.3322/caac.21166
[3] T. Matsuda, T. Marugame, K. Kamo, K. Katanoda, W. Ajiki, T. Sobue and Group Japan Cancer Surveillance Research, “Cancer Incidence and Incidence Rates in Japan in 2006: Based on Data from 15 Population-Based Cancer Registries in the Monitoring of Cancer Incidence in Japan (MCIJ) Project,” Japanese Journal of Clinical Oncology, Vol. 42, No. 2, 2012, pp. 139-147. http://dx.doi.org/10.1093/ jjco/hyr184
[4] R. A. Smith, D. Brooks, V. Cokkinides, D. Saslow and O. W. Brawley, “Cancer Screening in the United States, 2013: A Review of Current American Cancer Society Guidelines, Current Issues in Cancer Screening, and New Guidance on Cervical Cancer Screening and Lung Cancer Screening,” CA: A Cancer Journal for Clinicians, Vol. 63, No. 2, 2013, pp. 88-105. http://dx.doi.org/10.3322/caac.21174
[5] R. A. Smith, V. Cokkinides and O. W. Brawley, “Cancer Screening in the United States, 2009: A Review of Current American Cancer Society Guidelines and Issues in Cancer Screening,” CA: A Cancer Journal for Clinicians, Vol. 59, No. 1, 2009, pp. 27-41. http://dx.doi.org/10.3322/caac.20008
[6] J. Holst, O. Koskela and B. von Schoultz, “Endometrial Findings Following Curettage in 2018 Women According to Age and Indications,” Annales Chirurgiae et Gynaecologiae, Vol. 72, No. 5, 1983, pp. 274-277.
[7] T. Gredmark, S. Kvint, G. Havel and L. A. Mattsson, “Histopathological Findings in Women with Postmenopausal Bleeding,” British Journal of Obstetrics and Gynaecology, Vol. 102, No. 2, 1995, pp. 133-136. http://dx.doi.org/10.1111/j.1471-0528.1995.tb09066.x
[8] Y. C. Choo, K. C. Mak, C. Hsu, T. S. Wong and H. K. Ma, “Postmenopausal Uterine Bleeding of Nonorganic Cause,” Obstetrics and Gynecology, Vol. 66, No. 2, 1985, pp. 225-228.
[9] B. Gull, B. Karlsson, I. Milsom and S. Granberg, “Can Ultrasound Replace Dilation and Curettage? A Longitudinal Evaluation of Postmenopausal Bleeding and Transvaginal Sonographic Measurement of the Endometrium as Predictors of Endometrial Cancer,” American Journal of Obstetrics and Gynecology, Vol. 188, No. 2, 2003, pp. 401-408.
[10] E. Kondo, T. Tabata, Y. Koduka, K. Nishiura, K. Tanida, T. Okugawa and N. Sagawa, “What Is the Best Method of Detecting Endometrial Cancer in Outpatients? Endometrial Sampling, Suction Curettage, Endometrial Cytology,” Cytopathology, Vol. 19, No. 1, 2008, pp. 28-33. http://dx.doi.org/10.1111/j.1365-2303.2007.00509.x
[11] B. R. Kipp, F. Medeiros, M. B. Campion, T. J. Distad, L. M. Peterson, G. L. Keeney, K. C. Halling and A. C. Clayton, “Direct Uterine Sampling with the Tao Brush Sampler Using a Liquid-Based Preparation Method for the Detection of Endometrial Cancer and Atypical Hyperplasia: A Feasibility Study,” Cancer, Vol. 114, No. 4, 2008, pp. 228-235. http://dx.doi.org/10.1002/cncr.23636
[12] Y. Koga, M. Yasunaga, M. Kajikawa, E. Shimizu, R. Takamatsu, R. Kataoka, Y. Murase, Y. Sasajima, T. Kasamatsu, T. Kato, T. Onda, S. Ikeda, M. Ishikawa, K. Ishitani, H. Ohta and Y. Matsumura, “Novel Virtual Cytological Analysis for the Detection of Endometrial Cancer Cells Using Autoscan Fluoromicroscopy,” Cancer Science, Vol. 102, No. 5, 2011, pp. 1068-1075. http://dx.doi.org/10.1111/ j.1349-7006.2011.01903.x
[13] Y. Koga, M. Yasunaga, S. Katayose, Y. Moriya, T. Akasu, S. Fujita, S. Yamamoto, H. Baba and Y. Matsumura, “Improved Recovery of Exfoliated Colonocytes from Feces Using Newly Developed Immunomagnetic Beads,” Gastroenterology Research & Practice, Vol. 2008, 2008, Article ID: 605273. http://dx.doi.org/10.1155/2008/605273
[14] S. Grenier, M. Sandig, D. W. Holdsworth and K. Mequanint, “Interactions of Coronary Artery Smooth Muscle Cells with 3D Porous Polyurethane Scaffolds,” Journal of Biomedical Materials Research Part A, Vol. 89, No. 2, 2009, pp. 293-303. http://dx.doi.org/10.1002/jbm.a.31972
[15] W. M. Merritt, A. A. Kamat, J. Y. Hwang, J. BottsfordMiller, C. Lu, Y. G. Lin, D. Coffey, W. A. Spannuth, E. Nugent, L. Y. Han, C. N. Landen, A. M. Nick, R. L. Stone, K. Coffman, E. Bruckheimer, R. R. Broaddus, D. M. Gershenson, R. L. Coleman and A. K. Sood, “Clinical and Biological Impact of EphA2 Overexpression and Angiogenesis in Endometrial Cancer,” Cancer Biological Therapy, Vol. 10, No. 12, 2010, pp. 1306-1314. http://dx.doi.org/10.4161/cbt.10.12.13582
[16] S.-K. Wang and C. S. P. Sung, “Spectroscopic Characterization of Model Urea, Urethane Compound, and Diamine Extender for Polyurethane-Urea,” Macromolecules, Vol. 35, No. 3, 2002, pp. 877-882. http://dx.doi.org/10.1021/ ma011316+
[17] K. El-Sahwi, S. Bellone, E. Cocco, F. Casagrande, M. Bellone, M. Abu-Khalaf, N. Buza, F. A. Tavassoli, P. Hui, D. Ruttinger, D. A. Silasi, M. Azodi, P. E. Schwartz, T. J. Rutherford, S. Pecorelli and A. D. Santin, “Overexpression of EpCAM in Uterine Serous Papillary Carcinoma: Implications for EpCAM-Specific Immunotherapy with Human Monoclonal Antibody Adecatumumab (MT201),” Molecular Cancer Therapy, Vol. 9, No. 1, 2010, pp. 57-66. http://dx.doi.org/10.1158/1535-7163.MCT-09-0675
[18] E. Yilmaz, M. Koyuncuoglu, I. B. Gorken, E. Okyay, B. Saatli, E. C. Ulukus and U. Saygili, “Expression of Matrix Metalloproteinase-2 and Survivin in Endometrioid and Nonendometrioid Endometrial Cancers and Clinicopathologic Significance,” Journal of Gynecologic Oncology, Vol. 22, No. 2, 2011, pp. 89-96. http://dx.doi.org/10.3802/jgo.2011.22.2.89
[19] S. K. Weber, A. Sauerwald, M. Polcher, M. Braun, M. Debald, N. B. Serce, W. Kuhn, G. Brunagel-Walgenbach and C. Rudlowski, “Detection of Lymphovascular Invasion by D2-40 (Podoplanin) Immunoexpression in Endometrial Cancer,” International Journal of Gynecological Cancer, Vol. 22, No. 8, 2012, pp. 1442-1448. http://dx.doi.org/10.1097/IGC.0b013e318269139b
[20] T. Boren, Y. Xiong, A. Hakam, R. Wenham, S. Apte, Z. Wei, S. Kamath, D. T. Chen, H. Dressman and J. M. Lancaster, “MicroRNAs and Their Target Messenger RNAs Associated with Endometrial Carcinogenesis,” Gynecologic Oncology, Vol. 110, No. 2, 2008, pp. 206-215. http://dx.doi.org/10.1016/ j.ygyno.2008.03.023
[21] W. Wu, Z. Lin, Z. Zhuang and X. Liang, “Expression Profile of Mammalian microRNAs in Endometrioid Adenocarcinoma,” European Journal of Cancer Prevention, Vol. 18, No. 1, 2009, pp. 50-55. http://dx.doi.org/10.1097/CEJ.0b013e328305a07a
[22] T. K. Chung, T. H. Cheung, N. Y. Huen, K. W. Wong, K. W. Lo, S. F. Yim, N. S. Siu, Y. M. Wong, P. T. Tsang, M. W. Pang, M. Y. Yu, K. F. To, S. C. Mok, V. W. Wang, C. Li, A. Y. Cheung, G. Doran, M. J. Birrer, D. I. Smith and Y. F. Wong, “Dysregulated Micrornas and Their Predicted Targets Associated with Endometrioid Endometrial Adenocarcinoma in Hong Kong Women,” International Journal of Cancer, Vol. 124, No. 6, 2009, pp. 1358-1365. http://dx.doi.org/10.1002/ijc.24071
[23] E. Bignotti, M. Ragnoli, L. Zanotti, S. Calza, M. Falchetti, S. Lonardi, S. Bergamelli, E. Bandiera, R. A. Tassi, C. Romani, P. Todeschini, F. E. Odicino, F. Facchetti, S. Pecorelli and A. Ravaggi, “Diagnostic and Prognostic Impact of Serum HE4 Detection in Endometrial Carcinoma Patients,” British Journal of Cancer, Vol. 104, No. 9, 2011, pp. 1418-1425. http://dx.doi.org/10.1038/bjc.2011.109
[24] E. Kalogera, N. Scholler, C. Powless, A. Weaver, R. Drapkin, J. Li, S. W. Jiang, K. Podratz, N. Urban and S. C. Dowdy, “Correlation of Serum HE4 with Tumor Size and Myometrial Invasion In Endometrial Cancer,” Gynecologic Oncology, Vol. 124, No. 2, 2012, pp. 270-275. http://dx.doi.org/10.1016/ j.ygyno.2011.10.025
[25] I. Mutz-Dehbalaie, D. Egle, S. Fessler, M. Hubalek, H. Fiegl, C. Marth and A. Widschwendter, “HE4 Is an Independent Prognostic Marker in Endometrial Cancer Patients,” Gynecologic Oncology, Vol. 126, No. 2, 2012, pp. 186-191. http://dx.doi.org/10.1016/j.ygyno.2012.04.022
[26] R. G. Moore, C. M. Miller, A. K. Brown, K. Robison, M. Steinhoff and G. Lambert-Messerlian, “Utility of Tumor Marker HE4 to Predict Depth of Myometrial Invasion in Endometrioid Adenocarcinoma of the Uterus,” International Journal of Gynecological Cancer, Vol. 21, No. 7, 2011, pp. 1185-1190. http://dx.doi.org/10.1097/IGC.0b013e3182229ad8
[27] R. Angioli, F. Plotti, S. Capriglione, R. Montera, P. Damiani, R. Ricciardi, A. Aloisi, D. Luvero, E. V. Cafa, N. Dugo, M. Angelucci and P. Benedetti-Panici, “The Role of Novel Biomarker HE4 in Endometrial Cancer: A Case Control Prospective Study,” Tumour Biology, Vol. 34, No. 1, 2013, pp. 571-576. http://dx.doi.org/10.1007/ s13277-012-0583-0

  
comments powered by Disqus

Copyright © 2018 by authors and Scientific Research Publishing Inc.

Creative Commons License

This work and the related PDF file are licensed under a Creative Commons Attribution 4.0 International License.