乳腺癌及癌前病变α-微管蛋白和γ-微管蛋白免疫荧光定量分析及其意义

目的 研究人乳腺癌前病变、原位癌及浸润性癌中α-微管蛋白和γ-微管蛋白表达量的变化及其意义,进一步探讨中心体异常与乳腺癌发生发展的关系。方法 采用流式细胞术免疫荧光定量分析方法,检测30例乳腺导管上皮不典型增生、30例乳腺导管内癌和30例浸润性导管癌的中心体α-微管蛋白和γ-微管蛋白表达水平,另30例正常乳腺组织作为对照组,并与免疫组化检测所得数据进行比较。结果 在正常乳腺组织、导管上皮不典型增生、乳腺导管内癌和浸润性导管癌中,α-微管蛋白、γ-微管蛋白的过量表达程度不同（P=0．000）,浸润性导管癌组最高。随着导管上皮增生、不典型增生至进展为原位癌和浸润性导管癌的发展变化,α-微管蛋白、γ-微管蛋白表达量呈增高趋势,并与病理组织学上细胞增生和癌细胞生长分化程度明显相关。除不典型增生组和乳腺导管内癌组外,其余各组间差异均有统计学意义（P=0．001）。比较同组病例的α-微管蛋白、γ-微管蛋白表达程度,差异无统计学意义（P〈0．05）。结论 乳腺癌前病变中已有中心体蛋白的过度表达,在乳腺癌形成的早期关键性步骤中,中心体异常有可能在促使细胞过度增殖恶性转化的过程中起了重要作用。免疫荧光定量分析和免疫组化检测两种方法结合使用,可相互补充验证。

A quantitative immuno-fluorescence analysis of alpha-tubulin and gamma-tubulin proteins in precancerous lesion and carcinoma of the breast and their clinical significance

OBJECTIVE: In order to explore the correlation between the centrosome aberration and oncogenesis of the breast carcinoma, the expression of alpha-tubulin and gamma-tubulin proteins in breast precancerous lesions, ductal carcinoma in situ (DCIS) and invasive ductal carcinomas (IDC) was investigated. METHODS: Quantitative immunofluorescence analysis was performed for measuring centrosome proteins by FITC-labeled monoclonal anti-alpha and anti-gamma-tubulin antibodies in 90 cases with precancerous lesions, DCIS and IDC of the breast, respectively. Normal breast tissue from 30 cases were taken as control group. RESULTS: The average of positive (FITC-labeled) cells were 3.2, 11.6, 14.8, 23.1 (alpha-tubulin) and 3.3, 10.7, 14.5, 24.5 (gamma-tubulin) in four groups, respectively. There were significant differences of alpha-tubulin or gamma-tubulin expression among those groups (P = 0.000), respectively. The highest expression quantity was in IDC group and the lowest was in normal breast tissue. Their expression was significantly associated with cellular proliferation and differentiation. CONCLUSION: There is over-expression of the centrosome tubulin protein in the precancerous stage of the breast. The centrosome aberration may play an important role during the crucial early step of oncogenesis and it may promote the cellular cancerization or transformation into malignancy. Quantitative immuno-fluorescence analysis and immunohistochemistry can be complementary each other.