人精子细胞膜结合型透明质酸酶在乳腺癌的表达及临床病理意义

Expression of PH20 in primary and metastatic breast cancer and its pathological significance

目的 研究人精子细胞膜结合型透明质酸酶(PH20)在人原发、转移乳腺癌中的蛋白及mRNA表达,探讨其在肿瘤转移中的作用。方法 合成两个高抗原性人PH20多肽,免疫于兔皮下,检测、鉴定、纯化抗PH20抗体。应用免疫组织化学抗生物素蛋白-生物素复合物(ABC)法检测53例乳腺癌原发及淋巴结转移灶PH20的表达;Western印迹法检测5例新鲜乳腺癌组织PH20的表达。合成两条PH20寡核苷酸探针,应用原位杂交检测乳腺组织芯片,包括28例乳腺癌组织,5例乳腺癌旁组织,15例正常乳腺组织。结果 PH20免疫组织化学示:正常乳腺组织无表达(0/3),53例乳腺癌中31例呈阳性表达(58.4％),其中乳腺癌不伴有淋巴结转移阳性率48.2％,乳腺癌伴有淋巴结转移原发癌70.8％,转移癌83.3％。Western印迹法示5例乳腺癌均有PH20相应阳性条带的表达。寡核苷酸探针原位杂交显示75％(21/28)乳腺癌中有PH20mRNA表达。17例正常乳腺组织中2例乳腺小叶呈弱阳性表达。结论 PH20的表达似乎与乳腺癌的浸润转移行为呈正相关,提示透明质酸酶可能消化乳腺癌周围组织,并释放纤维母细胞生长因子-2而促进其肿瘤生长和转移。

Objective To evaluate the expression pattern of PH20 in primary and metastatic breast cancer and its relationship to tumor metastatic potential. Methods Anti-PH20 antibody was synthesized by injection of conjugated human PH20 peptides into rabbits. Immunohistochemical study was performed on 53 cases of human breast cancer. Western blot was used to detect PH20 expression in 5 cases of breast cancer with available fresh tissue. Two oligonucleotide probes were prepared for in-situ hybridization using breast tissue microarray . Results Normal breast tissue did not express PH20 (0/3) , while 58. 4% (31/53) of breast cancer cases did. The highest expression rate was found in metastatic foci in regional lymph nodes (83. 3% ) ,followed by primary breast cancer tissue in cases with lymph node secondaries (70. 8% ). The breast cancer cases with no any metastasis had an expression rate of 48. 2% . The immunohistochemical staining results were further confirmed by Western blotting. In-situ hybridization showed PH20 RNA in 75% of the breast cancer tissue (21/28). Two of the 17 cases of normal breast tissue showed weak expression in some ductolobular units. Conclusions The expression of PH20 has a positive correlation with metastatic potential in breast cancer. It is possible that PH20 may play an important role in the invasive growth and metastasis of breast cancer cells, via mechanisms such as digestion of surrounding stromal tissue and release of FGF-2.