Comparing the efficacy of routine H&E staining and cytokeratin immunohistochemical staining in detection of micro-metastasis on serial sections of dye-mapped sentinel lymph nodes in colorectal carcinoma


1 Department of Pathology-Acquired Immunodeficiency Research Center, Isfahan University of Medical Sciences, Isfahan, Iran

2 Department of General Surgery, Isfahan University of Medical Sciences, Isfahan, Iran

3 Department of Pathology, School of Medicine, Arak University of Medical Sciences, Arak, Iran


Background: The significance of techniques used for detecting micro-metastasis (MM) or isolated tumor cells (ITCs) is a controversial issue among investigators. We evaluated the different techniques used on sentinel lymph node (SLN) to detect MM/ITCs.
Materials and Methods: Ninety-one SLNs of 15 patients underwent serial section with 100 ΅m interval. In each level, two sections were prepared. One section was stained with H&E and another with anti-cytokeratin antibody (immunohistochemistry). Then the sections were evaluated for detecting MM/ITCs. Results were analyzed by chi-square test.
 1656 sections of 91 SLNs of 15 patients were evaluated by a pathologist; MM was found in 1 and ITCs in 1 case. Overall, 2 out of 15 cases (13.3% of the patients) showed MM/ITCs by IHC staining. So, serial section along with using IHC was superior than serial section and routine H&E staining. But it did not affect the 5-year survival of the patients (P = 0.47).
 Using the combined techniques of serial section and IHC staining could up-stage 13.3% of colon cancer patients who were lymph node negative. In other studies with different combination of serial section, IHC staining, and PCR, investigators were able to find MM/ITCs in 3-39% of the cases. In our study, although serial section and IHC staining could up-stage 13.3% of patients, it could not affect the 5-year survival of the patients.


Di Giacomo M, Altomare D, Guanti G. Micrometatstasis in colorectal cancer. Acta Chir Iugosl 2002;49:63-6.  Back to cited text no. 1
Schulze T, Bembenek A, Schlag PM. Sentinel lymph node biopsy progress in surgical treatment of cancer. Langenbecks Arch Surg 2004;389:532-50.  Back to cited text no. 2
Schlag PM, Bembenek A, Schulze T. Sentinel node biopsy in gastrointestinal-tract cancer. Eur J Cancer 2004;40:2022-32.  Back to cited text no. 3
Yuan HY, Cheng FL, Wei ZZ, Yang GL, Chen JK. Clinical significance of detecting lymph node micrometastasis of colorectal cancer by reverse transcriptase-polymerase chain reaction (RT-PCR. Ai Zheng 2004;23:1069-73.  Back to cited text no. 4
Palma RT, Waisberg J, Bromberg SH, Simão AB, Godoy AC. Micrometastasis in regional lymph nodes of extirpated colorectal carcinoma: Immunohistochemical study using anti-cytokeratin antibodies AE1/AE3. Colorectal Dis 2003;5:164-8.  Back to cited text no. 5
Pozza E, Ascanelli S, Turini A, Tonini G, Carcoforo P, Navarra G. Impact of the sentinel lymph node in the staging of colorectal carcinoma. Chir Ital 2002;54:659-65.  Back to cited text no. 6
Wang FL, Pan ZZ, Wan DS. Mapping the sentinel lymph node ex vivo and finding in micrometastasis by CK-immunostaining in carcinoma of colon and rectum. Zhonghua Wai Ke Za Zhi 2005;43:994-7.  Back to cited text no. 7
Bembenek A, Schneider U, Gretschel S, Ulmer C, Schlag PM. Optimization of staging in colon cancer using sentinel lymph node biopsy. Chirurg 2005;76:58-67.  Back to cited text no. 8
Bembenek A, Schneider U, Gretschel S, Fischer J, Schlag PM. Detection of lymph node micrometastases and isolated tumor cells in sentinel and nonsentinel lymph nodes of colon cancer patients. World J Surg 2005;29:1172-5.  Back to cited text no. 9
Dragan R, Nebojsa M, Dejan S, Ivan P, Dragos S, Damir J, et al. Clinical application of sentinel lymph node biopsy for staging, treatment and prognosis of colon and gastric cancer. Hepatogastroenterology 2009;56:1606-11.  Back to cited text no. 10
Wasif N, Faries MB, Saha S, Turner RR, Wiese D, McCarter MD, et al. Predictors of occult nodal metastasis in colon cancer: Results from a prospective multicenter trial. Surgery 2010;147:352-7.   Back to cited text no. 11
D'Armento G, Daniele L, Mariani S, Ottaviani D, Mussa A, Cassoni P, et al. Added value of combined gene and protein expression of CK20 and CEA in non-macroscopically involved lymph nodes of colorectal cancer. Int J Surg Pathol 2009;17:93-8.   Back to cited text no. 12
Lim SJ, Feig BW, Wang H, Hunt KK, Rodriguez-Bigas MA, Skibber JM, et al. Sentinel lymph node evaluation does not improve staging accuracy in colon cancer. Ann Surg Oncol 2008;15:46-51.   Back to cited text no. 13
Kelder W, Braat AE, Karrenbeld A, Grond JA, De Vries JE, Oosterhuis JW, et al. The sentinel node procedure in colon carcinoma: A multi-centre study in The Netherlands. Int J Colorectal Dis 2007;22:1509-14.  Back to cited text no. 14
Bembenek AE, Rosenberg R, Wagler E, Gretschel S, Sendler A, Siewert JR, et al. Sentinel lymph node biopsy in colon cancer: A prospective multicenter trial. Ann Surg 2007;245:858-63.  Back to cited text no. 15
Kelder W, van den Berg A, van der Leij J, Bleeker W, Tiebosch AT, Grond JK, et al. RT-PCR and immunohistochemical evaluation of sentinel lymph nodes after in vivo mapping with Patent Blue V in colon cancer patients. Scand J Gastroenterol 2006;41:1073-8.  Back to cited text no. 16
Thomas KA, Lechner J, Shen P, Waters GS, Geisinger KR, Levine EA. Use of sentinel node mapping for cancer of the colon: 'To map or not to map." Am Surg 2006;72:606-12.  Back to cited text no. 17
Park JS, Choi GS, Kim HJ, Park SY, Park YJ, Lee SW, et al. The feasibility of an ex-vivo sentinel lymph mapping using preoperative radioisotope injection in cases of extraperitoneal rectal cancer. J Korean Soc Coloproctol 2011;27:83-9.   Back to cited text no. 18
Lips DJ, Koebrugge B, Liefers GJ, van de Linden JC, Smit VT, Pruijt HF, et al. The influence of micrometastases on prognosis and survival in stage I-II colon cancer patients: The enroute Å Study. BMC Surg 2011;11:11.  Back to cited text no. 19
Braat AE, Pol RA, Oosterhuis JW, de Vries JE, Mesker WE, Tollenaar RA. Excellent prognosis of node negative patients after sentinel node procedure in colon carcinoma: A 5-year follow-up study. Eur J Surg Oncol 2014;40:747-55.  Back to cited text no. 20