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目的:建立更敏感、更准确的多标记逆转录-聚合酶链式反应(reverse transcriptase-polymerase chainreaction,RT-PCR)方法检测常规病理检查淋巴结阴性的宫颈癌淋巴结微转移。方法:选择行子宫根治术及盆腔淋巴结清扫术的18例早期宫颈癌患者162枚淋巴结,应用 RT-PCR、常规病理方法分别检测,RT-PCR的标记物为细胞角蛋白(CK)19 mRNA,人端粒酶逆转录酶(hTERT)mRNA,如其一项阳性则为淋巴结微转移。结果:1.逆转录-聚合酶链式反应:在所有的肿瘤组织均有 CK19 mRNA 及94.44%(16/18)的肿瘤组织中有 hTERT mRNA 表达。10枚正常淋巴结两者均无表达。盆腔淋巴结同时进行 CK19mRNA,hTERTmRNA 的 RT-PCR 及组织学检查,在组织学检查发现淋巴结转移的7枚中,CK19mRNA 均表达,其中6枚 hTERTmRNA 表达,在组织学未发现转移的155个淋巴结中,19枚 CK19mRNA 为阳性,18枚 hTERT mRNA 为阳性,在 CK19mRNA 和 hTERT mRNA 的 RT-PCR 方法中,共有4例22枚淋巴结发现转移。RT-PCR 方法检测宫颈癌的转移比组织学检查更为敏感(p<0.001)。结论:1.本研究表明:在早期宫颈癌淋巴结的微转移率可达14.19%(22/155);2.CK19 mRNA 及 hTERT mRNA 多标记的 RT-PCR 方法检测淋巴结的微转移比常规的病理检查更为敏感和准确。3.RT-PCR 方法在筛选组织学检查淋巴结阴性而有高度复发危险性的患者具有实用价值,并可指导治疗,判断预后。
OBJECTIVE: To establish a more sensitive and accurate reverse transcriptase-polymerase chain reaction (RT-PCR) method to detect lymph node micrometastasis in lymph node-negative cervical lymph nodes. Methods: Totally 162 lymph node specimens from 18 patients with early cervical cancer who underwent hysterectomy and pelvic lymphadenectomy were detected by RT-PCR and routine pathological methods respectively. The markers of RT-PCR were cytokeratin 19 mRNA, Human telomerase reverse transcriptase (hTERT) mRNA, if a positive lymph node micrometastasis. Reverse transcriptase-polymerase chain reaction: There was CK19 mRNA in all tumor tissues and hTERT mRNA expression in 94.44% (16/18) tumor tissues. None of the 10 normal lymph nodes were expressed. In the pelvic lymph nodes, CK19mRNA and hTERTmRNA were simultaneously detected by RT-PCR and histological examination. CK19mRNA was expressed in 7 of the lymph node metastases detected by histological examination. Of them, 6 hTERTmRNA was expressed in 155 lymph nodes without metastasis, Nineteen CK19 mRNAs were positive and 18 hTERT mRNAs were positive. In the RT-PCR method of CK19 mRNA and hTERT mRNA, a total of 22 cases of lymph node metastases were found. Detection of cervical cancer by RT-PCR was more sensitive than histology (p <0.001). The results of this study showed that the micrometastasis rate of lymph node in early cervical cancer was up to 14.19% (22/155) .2. The multi-labeled RT-PCR of CK19 mRNA and hTERT mRNA detected the lymph node micrometastasis more than the conventional pathology Check more sensitive and accurate. The RT-PCR method has practical value in the screening of patients with negative lymph node histology and high risk of recurrence, and can guide the treatment and prognosis.