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逆转录聚合酶链反应(Rt-PCR)在口腔鳞状细胞癌患者切开活检后检测播散癌细胞中的应用。

Use of Rt-PCR in detecting disseminated cancer cells after incisional biopsy among oral squamous cell carcinoma patients.

作者信息

Ramani Pratibha, Thomas George, Ahmed Shaheen

机构信息

Department of Oral And Maxillofacial Pathology, Saveetha Dental College &Hospital, Chennai-77, Tamil Nadu, India.

出版信息

J Cancer Res Ther. 2005 Apr-Jun;1(2):92-7. doi: 10.4103/0973-1482.16708.

Abstract

PURPOSE

Tumour metastasis is the most clinically significant and enigmatic aspect of tumour behavior and is an unequivocal hallmark of malignancy. Until recent years little has been known about the transportation phase of vascular dissemination during biopsy, because of the technical difficulties in demonstrating circulating cancer cells.

AIMS

This study examined whether cancer cell dissemination results from incisional biopsy in the peripheral blood by using Cytokeratin 19(CK-19) as the marker for Reverse Transcriptase-Polymerase Chain Reaction (RT-PCR). In-house recipes without utilizing kits were employed to extract genomic and total RNA to make the procedure user friendly.

MATERIALS AND METHODS

The study population consisted of n=10 patients who were clinically diagnosed for oral squamous cell carcinoma and who had not undergone any previous biopsies. 5 patients who were to undergo incisional biopsies for benign conditions served as controls. 5 ml of blood aspirates were collected before and within 15 minutes after incisional biopsy. CK-19 gene and a positive control gene beta actin were isolated to confirm the primers. Using the total RNA, RT-PCR was performed for beta actin and Ck 19 gene expression.

RESULTS

Rt-PCR did show any expression for the CK-19 gene.

CONCLUSION

In conclusion there was no evidence of dissemination of cancer cells in our study and the patients are on a regular follow up for the past one and half years. But larger sample size should be examined to make the procedure a diagnostic tool for cancer metastasis.

摘要

目的

肿瘤转移是肿瘤行为中临床上最重要且最神秘的方面,是恶性肿瘤明确无误的标志。直到近年来,由于在证明循环癌细胞方面存在技术困难,对于活检期间血管播散的运输阶段了解甚少。

目的

本研究通过使用细胞角蛋白19(CK-19)作为逆转录聚合酶链反应(RT-PCR)的标志物,来检测切开活检是否会导致外周血中癌细胞播散。采用不使用试剂盒的自制方法提取基因组RNA和总RNA,以使该过程便于使用。

材料与方法

研究人群包括n = 10例临床诊断为口腔鳞状细胞癌且此前未接受过任何活检的患者。5例因良性疾病接受切开活检的患者作为对照。在切开活检前及活检后15分钟内采集5毫升血样。分离CK-19基因和阳性对照基因β-肌动蛋白以确认引物。使用总RNA对β-肌动蛋白和Ck 19基因表达进行RT-PCR。

结果

RT-PCR未显示CK-19基因有任何表达。

结论

总之,在我们的研究中没有癌细胞播散的证据,并且在过去一年半中对患者进行了定期随访。但应检查更大的样本量,以使该方法成为癌症转移的诊断工具。

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