本文選題：循環(huán)腫瘤細(xì)胞　切入點(diǎn)：肝癌　出處：《第二軍醫(yī)大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

【摘要】：【研究目的】目前循環(huán)腫瘤細(xì)胞(circulating tumor cell,CTC)的檢測(cè)方法多受限于特異性或者準(zhǔn)確的細(xì)胞表面標(biāo)記,由于這些特異的細(xì)胞表面標(biāo)記只在很小的一部分細(xì)胞群體中表達(dá),因此,通過(guò)這類方法檢測(cè)到的CTC只占血液中循環(huán)的腫瘤細(xì)胞的很小一部分。在這篇文章中,我們發(fā)明了一種全新的CTC檢測(cè)技術(shù),該技術(shù)是基于檢測(cè)腫瘤細(xì)胞所共有的生物特性,即細(xì)胞核漿比,而不是生物標(biāo)記,如EpCAM等,因此,可以檢測(cè)到更大的CTC群體,提升CTC檢測(cè)的靈敏度,使檢測(cè)效果具有更高的穩(wěn)定性�！狙芯糠椒ā渴紫�,我們用量化成像流式Flow Sight檢測(cè)肝癌患者外周血中的循環(huán)細(xì)胞,發(fā)現(xiàn)了一個(gè)特殊的單核細(xì)胞群體,這個(gè)細(xì)胞群體為CD45-的細(xì)胞,并且相較于其他細(xì)胞,具有更高的細(xì)胞核漿比,具有分裂能力。我們認(rèn)為其為循環(huán)腫瘤細(xì)胞,并且定義為高核漿比細(xì)胞(high karyoplasmic ratios,HKR cells)。隨后,我們用10例HCC伴發(fā)鏡下癌栓(microvascular invasion,MVI)的病人、5例正常人的外周血進(jìn)行進(jìn)一步檢測(cè),以此來(lái)建立外周血HKR細(xì)胞數(shù)目與鏡下癌栓出現(xiàn)之間的量化關(guān)系,為了增加實(shí)驗(yàn)的準(zhǔn)確性,同時(shí)標(biāo)記了CD45,EpCAM,DAPI標(biāo)記。然后,我們隨機(jī)選取了12例正常人,12例非肝癌病人(肝硬化等),52例肝癌病人(18例MVI0期的病人,18例MVI1期的病人,16例MVI2期的病人)的外周血進(jìn)一步檢測(cè)建立的量化關(guān)系,并以此來(lái)確立應(yīng)用外周血中HKR細(xì)胞數(shù)目來(lái)預(yù)測(cè)MVI出現(xiàn)的模型。并且,利用受試者工作特征曲線(Reciever Operation Curves,ROC)評(píng)價(jià)了HKR細(xì)胞數(shù)目診斷MVI出現(xiàn)和診斷肝癌發(fā)生的靈敏度和特異度,同時(shí),我們對(duì)入組的腫瘤病人進(jìn)行了最長(zhǎng)為期一年的術(shù)后隨訪,以確定我們所建立的方法在判斷腫瘤預(yù)后方面的臨床意義。最后,我們隨機(jī)選取了42名隨機(jī)病人和52名肝癌病人的外周血,來(lái)進(jìn)一步檢測(cè)我們所建立模型的準(zhǔn)確性�！緦�(shí)驗(yàn)結(jié)果】在肝癌病人中,運(yùn)用我們的方法檢測(cè)到的外周血HKR細(xì)胞數(shù)目有56.2±23.8個(gè)/100,000個(gè)單核細(xì)胞,該值遠(yuǎn)大于非腫瘤組的病人(7.6±1.2/100,000)。在肝癌病人中,伴發(fā)MVI和不伴發(fā)MVI的病人,外周血中檢測(cè)到的HKR細(xì)胞的數(shù)目也是有顯著差異。通過(guò)受試者工作特征曲線分析得知,閾值是21.8/100,000,曲線下面積大于傳統(tǒng)的檢測(cè)方法(比如依賴CD45和EpCAM標(biāo)記法)。這些結(jié)果表明這種新型的CTC檢測(cè)技術(shù)相對(duì)于傳統(tǒng)的方法,具有更高的敏感性和可靠性�！窘Y(jié)論】在這項(xiàng)研究中,基于外周血CTCs的核漿比,我們開(kāi)發(fā)了一種利用成像流式細(xì)胞儀,檢測(cè)量化外周血中CTCs的方法,并對(duì)這一方法進(jìn)行了驗(yàn)證。該技術(shù)方法依賴于檢測(cè)所有腫瘤細(xì)胞所共有的細(xì)胞形態(tài)學(xué)特征-核漿比,而不是依賴特異性抗體或特異的細(xì)胞表面標(biāo)志物,相比于傳統(tǒng)的依賴細(xì)胞表面標(biāo)記的CTCs檢測(cè)技術(shù),其具有更高的靈敏度,這表明它是CTC的檢測(cè)和監(jiān)測(cè)的一種更有效的方法。
[Abstract]:[objective] at present, the detection of circulating tumor cells is limited by specific or accurate cell surface markers, because these specific cell surface markers are expressed in a small number of cell populations. In this article, we developed a new technique for CTC detection, which is based on the biological properties common to tumor cells. That is, the ratio of nucleus to cytoplasm, rather than biomarkers, such as EpCAM, so that a larger CTC population can be detected, the sensitivity of CTC detection can be increased, and the detection effect will be more stable. [research method] first of all, We used quantitative imaging flow Flow Sight to detect circulating cells in peripheral blood of patients with liver cancer, and we found a special monocyte population of CD45- cells with a higher nuclear / cytoplasmic ratio than other cells. It is considered to be a circulating tumor cell and is defined as a high karyoplasmic cytoplasmic ratio (karyoplasmic) of HKR cells. Subsequently, we examined the peripheral blood of 5 normal controls in 10 patients with HCC accompanied by microvascular invasionus. The quantitative relationship between the number of peripheral blood HKR cells and the appearance of tumor thrombus under microscope was established. In order to increase the accuracy of the experiment, CD45 HKR was labeled with EpCAMU DAPI. We randomly selected 12 non-HCC patients (52 liver cancer patients with liver cirrhosis, 18 patients with MVI0 stage, 18 patients with MVI1 phase and 16 patients with MVI2 phase) to establish a quantitative relationship. The model of predicting the occurrence of MVI was established by using the number of HKR cells in peripheral blood. Furthermore, the sensitivity and specificity of the number of HKR cells in the diagnosis of MVI and the occurrence of HCC were evaluated by using the operating characteristic curve of the subjects. We followed up oncology patients for up to one year to determine the clinical significance of the established method in judging the prognosis of the tumor. We randomly selected the peripheral blood of 42 random patients and 52 patients with liver cancer to further test the accuracy of our model. The number of HKR cells in peripheral blood detected by our method was 56.2 鹵23.8 / 100000 monocytes, which was much higher than that in non-tumor patients, 7.6 鹵1.2 / 1000000.In patients with liver cancer, patients with MVI and those without MVI, The number of HKR cells detected in peripheral blood was also significantly different. The threshold is 21.8 / 100000, and the area under the curve is larger than the traditional detection methods (such as relying on CD45 and EpCAM markers). These results show that this new CTC detection technique is relative to the traditional methods. [conclusion] in this study, based on the nucleo-cytoplasmic ratio of peripheral blood CTCs, we developed a method for quantifying CTCs in peripheral blood by imaging flow cytometry. This method depends on the nuclear / cytoplasmic ratio, which is common to all tumor cells, rather than on specific antibodies or specific cell surface markers. Compared with the traditional CTCs detection technique which relies on cell surface marker, it has higher sensitivity, which indicates that it is a more effective method for detection and monitoring of CTC.
【學(xué)位授予單位】：第二軍醫(yī)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R735.7

【參考文獻(xiàn)】

相關(guān)期刊論文 前1條

1 吳孟超;湯釗猷;劉彤華;叢文銘;步宏;陳杰;董輝;朱玉瑤;馮龍海;陳駿;;原發(fā)性肝癌規(guī)范化病理診斷指南(2015年版)[J];臨床肝膽病雜志;2015年06期

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