贲门癌自体荧光的实验研究

Autofluorescence laboratory study of carcinoma of gastric cardia

目的探讨贲门癌组织显微自体荧光特征及作为自体荧光成分的Ⅲ、Ⅳ型胶原在贲门癌组织与正常胃组织中分布的不同。方法采用激光扫描共聚焦显微镜以氩离子（Ar^＋）激光（Ex=488nm）和氦氖（He-Ne）激光（Ex=543nm）为激发光的双通道法对20例贲门癌手术标本进行自体荧光图像分析；采用免疫组化技术，半定量计分法分析Ⅲ、Ⅳ型胶原在正常胃组织和贲门癌组织中的分布差异。结果贲门癌组织自体荧光信号与正常胃组织各层相比均显著减弱（P〈0．01）。正常胃组织的Ⅳ型胶原抗体反应呈强阳性，贲门癌组织则明显减弱；丽Ⅲ型胶原在贲门癌间质中的抗体阳性反应较正常胃组织变化不明显。结论贲门癌组织与正常胃组织的显微自体荧光无论在形态、颜色、分布，还是在荧光强度上都存在巨大的差异。贲门癌组织比正常胃组织细胞外基质内Ⅳ型胶原显著减少，丽Ⅲ型胶原减少不明显；Ⅳ型胶原可能是胃底贲门黏膜细胞外基质自体荧光的主要来源。

Objective To investigate the microscopic autofluorescence characteristic of carcinoma of gastric eardia and compare the distribution of collagen type Ⅲ and Ⅳ as the component of autofluorescence in mucosa of normal and carcinoma of gastric eardia tissues. Methods A double-channel laser scanning eonfocal microscopy with Argon ion laser（excitation wavelength 488 nm） and Helium-Neon laser （excitation wavelength 543 nm） were used to detect the autofluoreseenee emitted from 20 surgical eases of carcinoma of gastric eardia and corresponding normal gastric corpus lissues. The autofluorescenee image of carcinoma of gastric eardia and normal gastric corpus was analysed and eompared to the HE stained picture. The distribution difference in extracellular matrix of two type.s collagen between normal and carcinoma tissues were measured with immunohistochemieal technique with monoclonal anti-collagen type Ⅲ and IV and semi-quantitative score method. Results The fluorescence intensity of carcinomatous tissues was markedly weaker than that of every layer of normal tissues （P 〈 0.01 ）. The immunoreaction of collagen type Ⅳ in normal tissue was strongly positive, in carcinomatous tissue was weak apparently. And the difference of immunoreaction of collagen type Ⅲ in normal and carcinomatous tissue was not apparent. Conclusions There are remarkable differences in the shape, color, distribution and fluorescence intensity of microscopic autofluorescenee between carcinoma of gastric eardia tissues and normal gastric corpus tissues. Compare to the normal tissue, the expression of collagen type Ⅳ of carcinomatous tissue in extracellular matrix is decreased significantly, while the change of collagen type Ⅲ is unapparent. And collagen type Ⅳ maybe the main source of laser-induced autofluorescence in cardiac cancer extracellular matrix.