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目的探讨X染色体灭活检测肿瘤细胞克隆来源在诊断头颈鳞癌颈淋巴结微转移中的作用。方法对20例临床N0M0头颈部鳞癌包括10例术后病理诊断明确的颈淋巴结微转移癌和10例可疑颈淋巴结微转移癌,通过组织显微切割和蛋白酶K消化技术获得肿瘤组织DNA,在限制性酶切和PCR扩增后观察肿瘤细胞雄激素受体在X染色体上的标志,明确X染色体灭活情况;通过比较原发癌和转移灶或可疑微转移灶细胞的检测结果鉴定转移部位细胞与原发癌肿瘤细胞的克隆同源性,进而对颈淋巴转移情况做出正确诊断。结果10例不同程度表达肿瘤细胞表面标志、病理诊断明确的颈淋巴结转移癌与其对应的原发癌均为单克隆来源,并且具有相同的X染色体灭活方式,提示两者之间具有相同的克隆来源,证明颈淋巴结转移癌来自原发癌;10例原发灶不同程度地表达表皮生长因子受体(epidermalgrowthfactorreceptor,EGFR)和角蛋白,但颈淋巴结内可疑微转移灶不表达EGFR和角蛋白的可疑颈淋巴结微转移癌,X染色体灭活分析发现6例可疑转移灶细胞与原发癌细胞具有克隆同源性,证实为颈淋巴结内微转移;其余4例两者间不具有相同的细胞克隆来源,排除淋巴结转移癌。结论采用X染色体灭活法检测肿瘤细胞的克隆来源对诊断头颈肿瘤颈淋巴结微转移具有很好的应用前景和潜在临床实用价值。
Objective To investigate the role of X chromosome inactivation in detecting micrometastasis of cervical lymph node in head and neck squamous cell carcinoma (NSCLC). Methods Twenty cases of clinical N0M0 head and neck squamous cell carcinoma including 10 cases of well-differentiated cervical lymph node micrometastasis and 10 cases of suspected cervical lymph node micrometastasis were obtained. Tissue DNA was obtained by tissue microdissection and proteinase K digestion. After restriction enzyme digestion and PCR amplification, the androgen receptor in tumor cells were observed on the X chromosome and the inactivation of X chromosome was clarified; the metastasis was identified by comparing the detection results of primary cancer and metastatic lymph nodes or suspicious micrometastasis cells Part of the cells and primary cancer cell cloning homology, and then make the correct diagnosis of cervical lymph node metastasis. Results 10 cases of tumor cells with different degrees of surface markers, pathologically diagnosed cervical lymph node metastasis and its corresponding primary cancer are monoclonal origin, and have the same way of X chromosome inactivation, suggesting that the same between the two clones It was proved that cervical cancer with lymph node metastasis originated from primary cancer. In 10 primary tumors, epidermal growth factor receptor (EGFR) and keratin were expressed to varying degrees, but suspicious micrometastases in cervical lymph node did not express EGFR and keratin Suspicious cervical lymph node micrometastases, X chromosome inactivation analysis found that 6 cases of suspicious metastatic cells and primary cancer cells have clonal homology confirmed as cervical lymph node micrometastasis; the other four cases do not have the same cell clone between the two Source, rule out lymph node metastases. Conclusion The use of X chromosome inactivation to detect the clonal origin of tumor cells has a good application prospect and potential clinical value in the diagnosis of cervical lymph node micrometastasis in head and neck cancer.