Abstract
Objective To develop an immunohistochemical assay for the diagnosis of cutaneous malignant melanoma (CMM) micrometastasis via blood and lymphatic vessels, and to evaluate its clinical significance. Methods Fifty-three patients (32 males and 21 females) histopathologically diagnosed as CMM were enrolled in this study. The patients were aged (61.2±8.4) years (range, 52- 72 years). Tissue specimens were obtained from the central area of tumor in each case , and also from removed lymph nodes in some cases. The average duration of follow-up was (65.00±5.68) months. During the follow-up, 17 patients died of the recurrence or metastasis of CMM, and 6 patients were lost to follow-up. The expressions of D2- 40, S100 and CD34 antigens in 53 tissue specimens were examined by immunohistochemical staining with three individual monoclonal antibodies, or by an immunohistochemical method using 2 two-antibody cocktails (D2-40/S100 and CD34/S100) and double-color chromogens in single tissue sections. Results Of the 53 patients, 30.19% (16/53) were positive for hematoxylin-eosin (HE) staining combined with immunohistochemical staining with individual monoclonal antibodies, and 49.06% (26/53) for the immunohistochemical method using two-antibody cocktails and double-color chromogens. Statistical differences were found in the positive rate between the two methods (χ2=3.94, P<0.05). Compared with patients without blood/lymph vessel tumor emboli, those with blood/lymph vessel tumor emboli showed higher lymph node metastasis rate (80.77%(21/26) vs. 37.04% (10/27), χ2=10.43, P<0.001), but lower five-year survival rate (42.31% (11/26) vs. 70.37%(19/27), χ2=4.25, P<0.05). Conclusions The immunohistochemical method with two-antibody cocktails is superior to HE staining combined with immunohistochemical staining with individual monoclonal antibodies in the detection of blood/lymph vessel tumor emboli. And blood/lymph vessel tumor emboli may be an important prognostic factor in patients with CMM. Key words: Melanoma; Lymphatic metastasis; S100 proteins; Antigens, CD34; D2-40; Immunohistochemical method with antibody cocktails
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