| Background Primary cutaneous malignant melanoma(PCMM)is a common skin tumor with high malignancy. Its incidence rate is low. With the social and environmental degradation, changes in lifestyle and the aging of the population, the incidence of melanoma increases year by year in recent years. There are about 2 million new cases each year. The tumor is highly malignant degree with earlier lymphatic or hematogenous metastasis, and the unsatisfied effect of treatment after metastasis resulted in high mortality rate, so early diagnosis and treatment of melanoma are significant to control the prognosis of the disease. Rho C(Ras homology C)protein is a member of Rho subfamily in the small molecule G protein superfamily and plays an important role in signal transduction. Cell proliferation, cytoskeleton, cell activity, tumor angiogenesis and cell matrix degradation are related to tumor invasion and metastasis.Rho C can influence these processes by regulating downstream target effector molecules. Research shows that the upregulation of Rho C expression in somekind of tumers is related to the invasion and transfer,such as in breast cancer, ovarian cancer, esophageal squamous cell carcinoma, pancreatic cancer, gastric cancer, liver cancer and other malignant tumors.ROCK(Rho associated coiled coil formingprotein kinase) is one of the downstream effectors of Rho C protein.As a member of serine / threonine protein kinase,combined with Rho protein family relying on GTP. It plays an important role in regulating variety cell biological processes, involved in the development process of liver cancer, gastric cancer, esophageal squamous cell carcinoma and other tumors. Research shows that Rho C and ROCK1 have important biological function in tumors, but whether they two are involved in the development process of melanoma still not be reported in China.As the similarity regulation characteristics of them,we study on the expression of Rho C and ROCK1 in cutaneous malignant melanoma and nevocytic nevus by using SP immunohistochemical method, observing their expression in different tissues and the association with clinical pathological factors,and try to investigate the function of Rho C and ROCK1 in the pathogenesis of malignant melanoma and the correlation of them.Objective To detect the expression of Rho C and ROCK1 in cutaneous malignant melanoma and nevus respectively and explore their relationship as well as function in the pathogenesis of malignant melanoma.Materials and methods 1.The source of the materials: Specimens were randomly selected from Pathology Department in First Affiliated Hospital of Zhengzhou Universitybetween 2007 to 2014.These 90 primary cutaneous melanocytic tumor(skin and lymphoid tissue paraffin block specimens)all have complete clinical and pathological data,including 60 cases of primary cutaneous malignant melanoma and 30 cases of nevocytic nevus. Analyze the clinical and pathological data,including age,sex,Clark classification and TNM stage and so on. 2.Method: Detected the expression of Rho C and ROCK1 in 60 cases of melanoma tissues and 30 cases of nevocytic nevus tissue by using immunohistochemical SP method.The concrete steps according to reagent box explanation. Using PBS instead of primary antibody as negative contrast.The positive contrast of Rho C and ROCK1 were provided by the company. 3.Statistical analysis: The data was analyzed by SPSS 19.0 statistical software.The difference was analysed by Chi-Square test, the correlation was analysed by Spearman correlation analysis.The inspection level is α=0.05.Results Expression of Rho C protein in cutaneous malignant melanoma and nevocytic nevus The positive expression of Rho C was mainly localized in cytoplasm and(or) cell membrane, showing brown or yellowish brown diffuse distribution. The positive rate of Rho C in malignant melanoma and nevocytic nevus were 70.00%, 16.67%. Rho C expression in melanoma and lymph node is positive,in nevocytic nevus is weak or negative.The difference between cutaneous malignant melanoma and nevus cell nevus has statistical significance(c2=22.801,P=0.000).2 Expression of ROCK1 protein in cutaneous malignant melanoma and nevocytic nevus The positive expression of ROCK1 was mainly localized in cytoplasm and(or) cell membrane, showing brown or yellowish brown diffuse distribution. The positive rate of ROCK1 in cutaneous malignant melanoma and nevocytic nevus were 81.67%, 13.33%. ROCK1 expression in melanoma and lymph node is positive,in nevocytic nevus is weak or negative.The difference between cutaneous malignant melanoma and nevus cell nevus has statistical significance(c2=38.575,P=0.000). 3 The relationship between expression of Rho C and pathological in cutaneous malignant melanoma Among 60 cases of cutaneous malignant melanoma,the positive rate of Clark I to III was 56.25%(18/32);the positive rate of Clark IV to V was 85.71%(24/28),the latter is much higher than the former(P < 0.05). The positive rate of TNM stage I-III A was 58.82%(20/34);Rho C positive rate of stage IIIB to V was 84.62%(22/26), significantly higher than that of I-IIIA(P < 0.05). The positive rate of the cases with distant lymph node metastasis was 76.92%(10/13). the positive rate of the cases without distant lymph node metastasis was 68.09%(32/47), the difference was not statistically significant(P>0.05). 4 The relationship between expression of ROCK1 and pathological in cutaneous malignant melanoma Among 60 cases of cutaneous malignant melanoma, ROCK1 positive rate of Clark I to III was 75.00%(24/32);the positive rate of Clark IV to V was 92.86%(25/28), significantly higher than the former(P < 0.05). The positive rate of TNM stage I-III A was 73.53%(25/34);ROCK1 positive rate of stage IIIB to V was 92.31%(24/26), significantly higher than that of I-IIIA(P < 0.05). The positive rate of thecases with distant lymph node metastasis was 84.61%(11/13),the positive rate of the cases without distant lymph node metastasis was 80.85%(38/47), the difference was not statistically significant(P>0.05). 5 The correlation between Rho C and ROCK1 protein The Spearman correlation analysis showed the expression of Rho C and ROCK1 had remarkable positive correlation.(r=0.348, P<0.05).Conclusion 1.The expression of Rho C and ROCK1 proteins in cutaneous malignant melanoma were significantly higher than nevocytic nevus, suggesting that the two molecules may become a potential marker of melanoma. 2.The expression of Rho C and ROCK1 proteins in cutaneous malignant melanoma were correlated with the tumor grade, stage and invasion, suggesting that the high expression of these two molecules may promote invasion and metastasis of cutaneous malignant melanoma. 3.Rho C expression was positively correlated with ROCK1, suggesting that these two molecules may play a synergistic role in the invasion and metastasis of melanoma. The high expression of Rho C can promote the development of cutaneous malignant melanoma by effecting ROCK1 expression. |
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