| IntroductionCutaneous melanoma is characterized with high malignancy, poor prognosis and various biological behaviors affected by many factors. The most important determinative factor of the melanoma prognosis is lymph node condition, which is also associated with the tumor thickness. However, no consensus has been reached on the regional lymph node clearance , little has been done on the molecular indicator criteria making and the reports in this area are rare. Recent evidence indicates that CD44, a multifunctional adhesion receptor involved in cell -cell as well as in cell - matrix interactions, plays an important role m local progression, metastasis of malignant tumors, and guidance of nesting. CD44 variant is closely related to the metastasis. Ki -67 , the non - histone nuclear proliferation antigen with two polypeptide chains of 395kDa and 345 kDa respectively, has expression only during proliferation phases of cell cycle. Ki -67 has been used to predict meta-static potential and overall prognosis. To explore the role of expression of CD44V5 and Ki - 67 in melanomas carcinogenesis and lymph node metastasis, we used immunohistochemistry for CD44V5 and Ki -67 expression determination and analysed the correlation between CD44 V5and Ki -67 expression and clinical pathological features .MaterialA total of 70 well - characterized cutaneous melanocytic neoplasms from Liaoning tumor hospital were studied, as obtained fromJan. 1990 to Feb. 2000. These specimens were procured as 55 primary tumors, 34 lymph node metastasis, 19 combined, and 12 controls. 45 males and 25 females, ranging in age from 17 to 83 years, whose lesions had arisen in various locations such as 59 at extremity, 2 at neck, 7 at torso, 2 of unknown origin . All the samples were reexam-ined by experienced pathologist, two cases were excluded from the certified malignant melanoma. PTNM staging were given depending on the criterion made by American Combined association of cancer, 1988 : No:36 , N,2:34; Mo:60, M,:10; stage I :2, stageII:11, stage IH : 47 , stage IV: 10. Tumor thickness was measured by ocular micrometer and classified into 5 grades; <0. 75mm: 2, 0. 76 - 1. 50; 4, 1.51-3.00mm: 12, 3.01 -4.50mm: 12, >4.5mm:25. Clark grading divided the samples into 5 groups: grade I ( epidermis) : 0, grade II ( papillary layer of dermis) : 5, grade M ( shallow reticular layer of dermis) : 16, gradeIV ( deep reticular layer of dermis) : 16, grade V ( subcutaneous fat layer) ; 18. Avidin - biotin - peroxidase complex ( S - P) kit. CD44V5 mouse to man monoclonal antibody. Ki - 67 mouse to man monoclonal antibody. Antigen retrieval. S - P kit.MethodS - P immunohistochemicl staining : 2 paraffin sections of each specimen were cut at 5 fi,m thickness . After dewaxed and rehydrated , they were stained by routine S - P method. Those cases with no reactivity whatsoever were regarded as negative, whereas immunoreactivity in the others was scored semiquantitatively ( as " 1 to 1.9 ( - ) ," " 2 to 2.9( + ) ," and2=3 ( ++ ) , etc. ) by assessment of intracellular melanosome . Statistics : The SPSS program package was used in aPC computer for basic statistical calculations. The detailed methods included x2 test ,Kendalls enumeration data correlation coefficient -r and Fishers exact test.ResultsCD44V5 and Ki - 67 expression were obvious among malignant melanocytic cells,but rare in normal cells . CD44V5 and Ki -67 expression between primary tumor and its lymph node metastasis had no statistical significance.I . CD44V5 was expressed both on the membrane and in the cytoplasm , but the former was stronger than the latter. Therefore, positive result was given depending on the membrane staining.1. CD44V5 and melanoma infiltrationPositive expression rate of grade I - Iff (this grading was based on the tumor thickness) was 83.33% , ; gradelV - V 83.78% ,and there was no statistical significance (p >0. 05) . Similiarly , positive expression rate of grade I - Iff ( Clark grading) was 94. 74% , ; gradeIV - V 82.35% ,and there was no st... |
