本文關(guān)鍵詞： NIR IR-80 良惡性細(xì)胞 甲狀腺癌 FNA　出處：《江蘇大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

【摘要】：目的:本研究首先根據(jù)乳腺正常上皮細(xì)胞株(MCF-10A)和乳腺癌細(xì)胞株(MCF-7和VDA-MB231)對近紅外(NIR)七甲川花菁染料IR-80不同攝取量導(dǎo)致的熒光強(qiáng)度差異性,篩選出染料濃度和拍攝參數(shù)等能夠鑒別良惡性細(xì)胞最佳實(shí)驗(yàn)條件;檢測IR-80染料孵育后甲狀腺癌和良性甲狀腺結(jié)節(jié)熒光強(qiáng)度,與傳統(tǒng)甲狀腺穿刺細(xì)胞學(xué)診斷結(jié)果比較,評估IR-80染色方法用于輔助鑒別甲狀腺結(jié)節(jié)細(xì)針穿刺(FNA)標(biāo)本良惡性可能性。方法:細(xì)胞系試驗(yàn):通過倍比稀釋配制四種濃度的IR-80染料應(yīng)用液(5μM、2.5μM、1.25μM和0.625μM),分別與人乳腺癌細(xì)胞株MCF-7、MDA-MB231和人正常乳腺上皮細(xì)胞株MCF-10A共同孵育,應(yīng)用熒光顯微鏡在NIR通道下(700-900nm)觀察每種細(xì)胞在0.5s、1s和2s曝光時(shí)間的熒光強(qiáng)度。然后用Image J圖像處理軟件計(jì)算平均熒光強(qiáng)度,分析NIR染料IR-80對腫瘤細(xì)胞的優(yōu)先聚集作用,篩選出最佳實(shí)驗(yàn)條件能夠區(qū)別良惡性細(xì)胞。甲狀腺FNA標(biāo)本研究:篩選2015年12月～2016年10月在南京軍區(qū)南京總醫(yī)院進(jìn)行甲狀腺結(jié)節(jié)細(xì)針穿刺診斷的病人標(biāo)本共120例,經(jīng)細(xì)胞學(xué)診斷陽性結(jié)果59例(甲狀腺癌59例),陰性結(jié)果61例(良性結(jié)節(jié))。利用細(xì)胞學(xué)試驗(yàn)篩選出的最佳條件觀察甲狀腺癌細(xì)胞和非癌細(xì)胞對IR-80染料的攝取情況,以細(xì)胞學(xué)分析為診斷金標(biāo)準(zhǔn),計(jì)算IR-80染色方法的靈敏度和特異度,利用ROC曲線評價(jià)IR-80染色方法的診斷價(jià)值。結(jié)果:細(xì)胞系試驗(yàn):當(dāng)曝光時(shí)間固定時(shí),MCF-7、VDA-VB231和MCF-10A三種細(xì)胞的NI R染色強(qiáng)度均隨NI R染料IR-80孵育濃度的降低而逐漸降低;當(dāng)IR-80孵育濃度固定時(shí),隨著曝光時(shí)間的增加,同一種細(xì)胞的NIR染料攝取增加不明顯;IR-80孵育濃度和曝光時(shí)間相同時(shí),乳腺癌細(xì)胞株MCF-7和MDA-MB231在NI R下的熒光強(qiáng)度高于正常乳腺上皮細(xì)胞MCF-10A。當(dāng)IR-80孵育濃度為1.25μM,不同曝光條件下觀察,腫瘤細(xì)胞(MCF-7和MDA-MB231)發(fā)出較強(qiáng)熒光,而非腫瘤細(xì)胞(MCF-10A)不被染色。篩選出IR-80應(yīng)用液濃度為1.25μM,曝光時(shí)間為1s為最佳實(shí)驗(yàn)參數(shù)鑒別良惡性細(xì)胞。甲狀腺細(xì)針穿刺(FNA)標(biāo)本研究:甲狀腺癌標(biāo)本中的平均熒光強(qiáng)度明顯強(qiáng)于對照組非甲狀腺癌細(xì)胞:甲狀腺癌細(xì)胞在NIR范圍清楚、均勻攝取熒光;良性標(biāo)本中的細(xì)胞包括良性甲狀腺濾泡上皮細(xì)胞、淋巴細(xì)胞和巨噬細(xì)胞等不發(fā)出熒光。IR-80染色方法檢測120例甲狀腺FNA標(biāo)本,有65例呈陽性結(jié)果,55例呈陰性結(jié)果,靈敏度為100%(59/59),特異度為90.1%(55/61);ROC曲線下面積為0.949,面積的標(biāo)準(zhǔn)誤為0.023,IR-80染色方法結(jié)果用于判斷FNA標(biāo)本良惡性有顯著意義(P=0.000)。結(jié)論:NIR七甲川花菁染料IR-80進(jìn)行活細(xì)胞染色時(shí),乳腺癌細(xì)胞攝取染料明顯高于正常乳腺上皮細(xì)胞,IR-80染料孵育濃度為1.25μM,曝光時(shí)間為1s可用于鑒別良惡性細(xì)胞。將細(xì)胞系試驗(yàn)篩選出的最佳參數(shù)用于甲狀腺FNA標(biāo)本時(shí),只有甲狀腺癌細(xì)胞被IR-80染色,而非癌細(xì)胞不被染色,說明IR-80具有優(yōu)先聚集腫瘤細(xì)胞作用,能夠鑒別甲狀腺FNA標(biāo)本良惡性。IR-80染色方法靈敏度和特異度較高,ROC曲線表明其診斷價(jià)值高,可能成為輔助鑒別臨床甲狀腺FNA標(biāo)本良惡性的新方法。
[Abstract]:Objective: This study first according to normal breast epithelial cell line (MCF-10A) and breast cancer cell lines (MCF-7 and VDA-MB231) on the near infrared (NIR) fluorescence intensity difference of seven methine cyanine dyes with different IR-80 intake leads to the selected dye concentration and shooting parameters to the optimal experimental conditions for differential diagnosis of benign and malignant cells; detection of IR-80 dye after incubation of thyroid carcinoma and benign thyroid nodule fluorescence intensity, compared with the traditional thyroid cytology diagnosis results, evaluation of IR-80 staining method for differential diagnosis of thyroid nodules by fine needle aspiration (FNA) specimens of benign and malignant possibility. Methods: cell lines by IR-80 dye test: application of liquid dilution with four concentration (5 M, 2.5 M, 1.25 M and 0.625 M, respectively) and human breast cancer cell lines MCF-7, MDA-MB231 and normal human breast epithelial cell line MCF-10A were incubated with fluorescence microscopy in NIR channel Under the observation of each cell (700-900nm) in 0.5s, 1s and 2S. The fluorescence intensity of exposure time to calculate the average fluorescence intensity and then use the Image J image processing software, analysis of NIR dye IR-80 on tumor cells preferentially aggregation, and selected the best experimental conditions to distinguish between benign and malignant thyroid cells. FNA study: from December 2015 to October 2016 were selected fine needle puncture in diagnosis of thyroid nodules in the General Hospital of Nanjing military region Nanjing patient samples were 120 cases, the positive cytological diagnosis results of 59 cases (59 cases of thyroid carcinoma (61 cases), negative results of benign nodules). The best conditions were screened by cytology test observation of IR-80 dye uptake in thyroid cancer and non cancer cells in cells, cytological analysis as the gold standard for diagnosis, calculate the sensitivity and specificity of IR-80 staining method, IR-80 staining method for evaluation of diagnostic value of using ROC curve values. Results: fine Cell test: when the exposure time is fixed, MCF-7 three, VDA-VB231 and MCF-10A cells NI R staining with NI R to reduce the strength of dye IR-80 incubating concentration decreased; when incubated with IR-80 concentration is fixed, with the increase of the exposure time, NIR dye uptake same cells did not increase significantly; IR-80 incubating concentration and exposure time, the fluorescence intensity of MCF-7 and MDA-MB231 in breast cancer cell line NI in R is higher than that of normal mammary epithelial cells when incubated with IR-80 MCF-10A. concentration of 1.25 M, were observed under different exposure conditions, tumor cells (MCF-7 and MDA-MB231) emit strong fluorescence, and non tumor cells (MCF-10A) don't be stained. Selected IR-80 application concentration is 1.25 M and the exposure time of 1s is the best experimental parameters in differential diagnosis of benign and malignant thyroid cells. Fine needle aspiration (FNA) specimens of thyroid carcinoma: the average fluorescence intensity was stronger than the control Group of non thyroid cancer cells: thyroid cancer cells in the range of NIR clear, uniform uptake of fluorescence; cells in benign specimens including benign thyroid follicular epithelial cells, lymphocytes and macrophages do not fluoresce.IR-80 staining method to detect 120 cases of thyroid FNA specimens, 65 cases showed positive results, 55 cases showed negative results, the sensitivity was 100% (59/59), the specificity was 90.1% (55/61); the area under the ROC curve was 0.949, the area of the standard error is 0.023. IR-80 staining results for judging FNA specimens of benign and malignant significant (P=0.000). Conclusion: NIR seven methine cyanine dyes IR-80 live cell staining, dye uptake in breast cancer cells significantly higher than that of normal mammary epithelial cells, IR-80 dye incubation concentration is 1.25 M and the exposure time of 1s can be used for differential diagnosis of benign and malignant cells. The optimal parameters of cell lines were screened for thyroid FNA specimens, only Thyroid cancer cells by IR-80 staining, and non cancer cells were not stained, indicating that IR-80 has priority to aggregation of tumor cells, can differentiate benign and malignant thyroid specimens of FNA.IR-80 staining method of sensitivity and specificity is high, ROC curve showed that the diagnostic value is high, may become auxiliary differential diagnosis of benign and malignant thyroid specimens of FNA new method.

【學(xué)位授予單位】：江蘇大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R736.1;R581

【相似文獻(xiàn)】

相關(guān)碩士學(xué)位論文 前1條

1 陳肖;近紅外七甲川花菁染料IR-80用于鑒別甲狀腺良惡性結(jié)節(jié)的實(shí)驗(yàn)研究[D];江蘇大學(xué);2017年

，

本文編號：1552627