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采用分子信标成像技术检测宫颈癌细胞中的生存素表达:宫颈癌诊断的新策略。

Detection of survivin expression in cervical cancer cells using molecular beacon imaging: new strategy for the diagnosis of cervical cancer.

机构信息

Department of Gynaecology and Obstetrics, First Affiliated Hospital of Medical School, Xi'an Jiaotong University, 710061 Xi'an, China.

出版信息

Eur J Obstet Gynecol Reprod Biol. 2011 Nov;159(1):204-8. doi: 10.1016/j.ejogrb.2011.06.038. Epub 2011 Jul 12.

Abstract

OBJECTIVE

Development of novel approaches for quantitative analysis of gene expression in intact tumour cells could provide new methods for the detection of cervical cancer. Molecular beacons (MBs) are single-stranded oligonucleotide probes that form stem-and-loop structures. Survivin, a member of the 'inhibitor of apoptosis' family of proteins, is highly expressed in cervical cancer. The aim of this study was to determine the feasibility of MBs targeting wild-type survivin in the diagnosis of cervical cancer.

STUDY DESIGN

MBs with oligonucleotide sequences complementing survivin mRNA and covalently linked with FITC or Cy3 were designed and synthesized. The specificity and sensitivity of survivin MBs were examined in human cervical cancer cell lines and smears from cervical cancer patients, and confirmed by reverse transcriptase polymerase chain reaction (RT-PCR), immunocytochemistry, Western blotting and the thinprep cytological test (TCT).

RESULTS

Both survivin MB-FITC and MB-Cy3 produced a strong fluorescent signal in cervical cancer cells, and the intensity was consistent with the results from RT-PCR, Western blotting and immunocytochemical staining. In the initial clinical cohort study, the sensitivity of survivin MBs was 61.37% (27/44) and the specificity was 72.72% (34/44); the sensitivities of Western blotting and TCT were 76.1% (32/42) and 68.19% (30/44), respectively. No significant difference in sensitivity was observed between MBs, Western blotting and TCT for different tumour grades and International Federation of Gynecology and Obstetrics (FIGO) stages.

CONCLUSIONS

Survivin MBs are specific and sensitive molecular probes for the detection of cervical cancer cells. They have great potential in the early detection and follow-up of patients with cervical cancer.

摘要

目的

开发新的方法用于定量分析完整肿瘤细胞中的基因表达,这可能为宫颈癌的检测提供新的方法。分子信标(MB)是形成茎环结构的单链寡核苷酸探针。凋亡抑制因子家族的成员 Survivin 在宫颈癌中高表达。本研究旨在确定针对野生型 Survivin 的 MB 用于宫颈癌诊断的可行性。

研究设计

设计并合成了与 Survivin mRNA 互补的寡核苷酸序列的 MB,并与 FITC 或 Cy3 共价连接。通过逆转录聚合酶链反应(RT-PCR)、免疫细胞化学、Western blot 和液基薄层细胞学检测(TCT),在人宫颈癌细胞系和宫颈癌患者的涂片上检查 Survivin MB 的特异性和敏感性,并进行了验证。

结果

Survivin MB-FITC 和 MB-Cy3 在宫颈癌细胞中均产生强烈的荧光信号,其强度与 RT-PCR、Western blot 和免疫细胞化学染色的结果一致。在初步的临床队列研究中,Survivin MB 的敏感性为 61.37%(27/44),特异性为 72.72%(34/44);Western blot 和 TCT 的敏感性分别为 76.1%(32/42)和 68.19%(30/44)。MB、Western blot 和 TCT 对不同肿瘤分级和国际妇产科联合会(FIGO)分期的敏感性无显著差异。

结论

Survivin MB 是检测宫颈癌细胞的特异性和敏感的分子探针。它们在宫颈癌患者的早期检测和随访中具有很大的潜力。

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