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目的了解XAPC7在肺腺癌细胞和组织中的表达及定位情况,为后续的功能研究提供线索。方法首先采用Western-blot技术检测XAPC7在HBE135-E6E7正常支气管上皮细胞株、A549肺腺癌细胞株、NCI-H446小细胞肺癌细胞株中的表达;再用细胞免疫荧光技术,以HBE135-E6E7和A549细胞为研究对象,用鼠抗人XAPC7单克隆抗体分析该蛋白的细胞定位情况;最后利用免疫组织化学法随机检测34例肺腺癌及相应癌旁组织中XAPC7蛋白的定位及表达。结果 Western-blot显示:与HBE135-E6E7相比,XAPC7在NCI-H446中表达明显降低(P<0.05),在A549中表达有所增加,但差异无统计学意义(P>0.05)。激光共聚焦显微镜观察发现XAPC7在A549中主要定位于细胞浆,胞核中少见,而在HBE135-E6E7中除胞浆外,胞核中也有较多分布。免疫组化显示:XAPC7在肺腺癌和肺鳞癌组织细胞的胞浆和胞核中均有表达。在34例肺腺癌中,癌组织胞浆中XAPC7蛋白的表达明显高于癌旁组织胞浆的表达(Z=-4.341,P=0.000),而在癌组织与癌旁组织的胞核中XAPC7蛋白的表达差异无统计学意义(Z=-0.167,P=0.867)。另外,XAPC7蛋白的表达与年龄、性别、肿瘤大小、组织分化和有无淋巴结转移无关(P均>0.05)。结论 XAPC7在肺腺癌细胞株中有表达,主要定位于胞浆。在肺腺癌组织和癌旁组织胞浆中的表达差异有统计学意义,为后续研究打下了基础。
Objective To understand the expression and localization of XAPC7 in lung adenocarcinoma cells and tissues and provide clues for the follow-up functional studies. Methods Western-blot was used to detect the expression of XAPC7 in HBE135-E6E7 normal bronchial epithelial cell line, A549 lung adenocarcinoma cell line and NCI-H446 small cell lung cancer cell line. The expression of XAPC7 in HBE135-E6E7 and A549 cells were used to study the cell localization of the protein by using the anti-human XAPC7 monoclonal antibody. Finally, the localization and expression of XAPC7 protein in 34 cases of lung adenocarcinoma and corresponding paracancerous tissues were detected by immunohistochemistry. Results Western-blot showed that the expression of XAPC7 in NCI-H446 was significantly lower than that in HBE135-E6E7 (P <0.05), but not in A549 (P> 0.05). Confocal laser scanning microscopy showed that XAPC7 mainly located in the cytoplasm of A549 cells, rare in the nucleus, but in HBE135-E6E7 in addition to the cytoplasm, the nucleus also more distribution. Immunohistochemistry showed that XAPC7 was expressed in the cytoplasm and nucleus of lung adenocarcinoma and lung squamous cell carcinoma tissues. In 34 cases of lung adenocarcinoma, the expression of XAPC7 protein in the cytoplasm of cancer tissue was significantly higher than that in the para-cancerous tissue (Z = -4.341, P = 0.000) There was no significant difference in the expression of XAPC7 protein (Z = -0.167, P = 0.867). In addition, the expression of XAPC7 protein was not associated with age, gender, tumor size, tissue differentiation and lymph node metastasis (all P> 0.05). Conclusion XAPC7 is expressed in lung adenocarcinoma cell lines and mainly located in the cytoplasm. There was significant difference in the expression of cytoplasm between lung adenocarcinoma and para-cancerous tissue, which laid the foundation for the follow-up study.