本文選題：食管腺癌 + Barrett’s食管�。� 參考：《天津醫(yī)科大學(xué)》2015年博士論文

【摘要】：目的①探討雌激素受體ERα、ERβ及其亞型在Barrett’s食管細胞系及食管腺癌細胞系細胞中的表達,并比較兩種細胞系的表達差異。②探討不同雌激素受體ERα和ERβ表達的食管腺癌細胞系對化療藥5-FU、順鉑、他莫昔芬的敏感性,并對兩組數(shù)據(jù)進行相關(guān)分析,以期發(fā)現(xiàn)化療敏感性與雌激素受體表達的關(guān)系。③探討5-FU、順鉑聯(lián)合他莫昔芬對不同雌激素受體表達細胞系細胞生長的影響。方法①對Barrett’s食管細胞系BAR-T、Ch TERT、Gi TERT、Qh TERT及食管腺癌細胞系OE19、OE33、SKGT4、OACP4C、FLO-1、OACM5.1進行培養(yǎng),培養(yǎng)至細胞融合度約70%時收割細胞、提取蛋白,用EZQ工具包測蛋白濃度。采用Western blotting方法測定雌激素受體ERα、ERβ及其亞型在不同食管細胞系細胞中的表達水平。②利用MTS法測定最佳的MTS觀察時間;建立各細胞系96小時生長曲線;根據(jù)細胞生長曲線決定96孔板每孔種植細胞的數(shù)量;對種植于96孔板OE19、OE33、SKGT4、OACP4C、FLO-1、OACM5.1等細胞系細胞分別予不同濃度5-FU、順鉑、他莫昔芬化學(xué)治療72小時,比較各細胞系對藥物的不同敏感性并將其結(jié)果與雌激素受體表達結(jié)果進行Spearman相關(guān)分析。③根據(jù)其雌激素受體表達的不同,我們選取雌激素受體ERβ高表達、ERα低表達的OE19及雌激素受體ERβ低表達、ERα高表達OACM5.1作為聯(lián)合化療對象。對選取的細胞系進行5-FU聯(lián)合他莫昔芬及順鉑聯(lián)合他莫昔芬聯(lián)合化學(xué)治療72小時,對其生長情況進行MTS檢測。結(jié)果①通過Western blotting檢測,Barrett’s食管細胞系細胞與食管腺癌細胞系細胞均不同程度地表達ERα或者ERβ,ERα表達以61k Da、90k Da亞型為主,ERβ以59k Da、51.5k Da及44.9k Da亞型為主。然而,與Brrett’s食管細胞系相比,食管腺癌細胞系細胞雌激素受體無論ERα或者ERβ表達水平均較高,而且在亞型ERβ44.9k Da表達上,兩組相比差異具有統(tǒng)計學(xué)意義(P0.05)。②最佳的MTS觀察時間為加MTS溶液后4小時內(nèi),以2小時或3小時觀察為最佳,而24小時則不適宜觀察。根據(jù)各細胞系96小時生長曲線,其每孔的適宜種植細胞數(shù)OE19、OE33、SKGT4應(yīng)為2000個每孔,OACP4C、FLO-1、OACM5.1應(yīng)為3000個每孔。各細胞系對5-FU、順鉑、他莫昔芬藥物的化療敏感性不同,對其不同的敏感性與雌激素受體表達行Spearman相關(guān)分析得知,在各細胞系中,5-FU、順鉑、他莫昔芬的敏感性與雌激素受體ERα61k Da、ERα90k Da、ERβ59k Da、ERβ51.5k Da、ERβ44.9k Da亞型表達無相關(guān)性,而雌激素受體亞型ERβ51.5k Da與他莫昔芬的敏感性呈負相關(guān)。③在雌激素受體ERβ高表達、ERα低表達細胞系OE19中,他莫昔芬能顯著增加順鉑對細胞的殺傷作用,而他莫昔芬聯(lián)合5-FU則無此作用。在雌激素受體ERβ低表達、ERα高表達細胞系OACM5.1中,他莫昔芬聯(lián)合順鉑或5-FU均不能增加其殺傷作用。結(jié)論①Barrett’s食管細胞系細胞與食管腺癌細胞系細胞均不同程度地表達ERα、ERβ及其亞型,然而食管腺癌細胞系細胞雌激素受體ERα、ERβ表達水平均較高,尤其是ERβ44.9k Da的表達具有統(tǒng)計學(xué)意義,提示我們可以將此亞型作為鑒別Barrett’s食管和食管腺癌的指標(biāo)之一。同時,我們在食管腺癌細胞系細胞觀察到ERα61k Da亞型的表達。②不同的食管腺癌細胞系細胞對5-FU、順鉑、他莫昔芬的化療敏感性是不同的。細胞對他莫昔芬的化療敏感性與雌激素受體ERβ51.5k Da的表達呈負相關(guān),提示雌激素受體某些亞型的表達與他莫昔芬化療敏感性密切相關(guān)。③在高雌激素受體β、低α表達的食管腺癌細胞系中,他莫昔芬與順鉑具有協(xié)同殺傷作用。在高雌激素受體α、低β表達的食管腺癌細胞系則無此作用。這提示我們,對于高雌激素受體β、低α表達食管腺癌患者可以嘗試他莫昔芬與順鉑聯(lián)合治療,以提高臨床療效。
[Abstract]:Objective: To investigate the expression of estrogen receptor ER alpha, ER beta and its subtypes in Barrett 's esophagus cell lines and esophageal adenocarcinoma cell lines, and to compare the differences in the expression of two cell lines. 2. The sensitivity of esophageal adenocarcinoma cell lines with different estrogen receptor ER alpha and ER beta expression to chemotherapy drug 5-FU, cisplatin and tamoxifen, and the number of two groups. To explore the relationship between chemosensitivity and the expression of estrogen receptor. (3) to explore the effect of 5-FU, cisplatin and tamoxifen on the growth of cell lines in different estrogen receptors. (1) Barrett 's esophageal cell line BAR-T, Ch TERT, Gi TERT, Qh TERT, and esophageal adenocarcinoma cell lines are OE19. OE33 -1, OACM5.1 were cultured to reaping cells, extracting protein and measuring protein concentration by EZQ toolkit. Western blotting method was used to determine the expression level of estrogen receptor ER alpha, ER beta and its subtypes in different esophageal cell lines. (2) the optimal MTS observation time was determined by MTS method, and each cell line 96 was established by MTS method. Hourly growth curve; determine the number of cells per hole of the 96 orifice according to the cell growth curve; the cell lines grown on the 96 orifice OE19, OE33, SKGT4, OACP4C, FLO-1, OACM5.1, and other cell lines were treated with different concentrations of 5-FU, cisplatin, and tamoxifen for 72 hours, and compared the different sensitivity of each cell line to the drug and the result and estrogen Spearman correlation analysis of the receptor expression results. (3) we select the high expression of estrogen receptor ER beta, low expression of ER alpha, low expression of ER alpha and low expression of ER beta in estrogen receptor, and ER alpha high expression OACM5.1 as a combined chemotherapy object. The growth situation of Finn was detected by MTS for 72 hours. Results (1) the expression of Barrett 's esophageal cell line cells and esophageal adenocarcinoma cell lines expressed ER alpha or ER beta in varying degrees by Western blotting. The expression of ER a was dominated by 61k Da, 90K Da subtype. Compared with the rrett 's esophageal cell line, the levels of both ER A and ER beta in the esophageal adenocarcinoma cell line were higher, and the difference between the two groups was statistically significant (P0.05) in the expression of the subtype ER beta 44.9k Da. The best time for MTS observation was to be best observed in 2 hours or 3 hours after the addition of MTS solution, and 24 small. According to the 96 hour growth curve of each cell line, the number of suitable planting cells per pore OE19, OE33, SKGT4 should be 2000 per pore, OACP4C, FLO-1, OACM5.1 should be 3000 per pore. The chemosensitivity of each cell line to 5-FU, cisplatin and tamoxifen is different, and its sensitivity to the expression of estrogen receptor is Spearman. The sensitivity of 5-FU, cisplatin and tamoxifen to the estrogen receptor ER alpha 61k Da, the ER alpha 90K Da, the ER beta 59K Da, the ER beta 51.5k, and the estrogen receptor subtype are negatively correlated with the sensitivity of tamoxifen. In cell line OE19, tamoxifen can significantly increase cisplatin's killing effect on cells, while tamoxifen combined with 5-FU without this effect. In the low expression of estrogen receptor ER beta, in the ER alpha high expression cell line OACM5.1, tamoxifen combined with cisplatin or 5-FU can not increase its killing effect. The cell line cells express ER alpha, ER beta and its subtypes in varying degrees. However, the expression level of estrogen receptor ER A and ER beta in the cell lines of the esophageal adenocarcinoma cell lines is higher, especially the expression of ER beta 44.9k Da, which suggests that we can use this subtype as one of the indicators to identify Barrett 's esophagus and esophageal adenocarcinoma. The expression of ER alpha 61k Da subtype was observed in the cells of the esophageal adenocarcinoma cell line. 2. The chemotherapeutic sensitivity of different esophageal adenocarcinoma cell lines to 5-FU, cisplatin and tamoxifen was different. The chemosensitivity of cells to tamoxifen was negatively correlated with the expression of the estrogen receptor ER beta 51.5k Da, suggesting the expression of some of the estrogen receptor subtypes and the expression of some of the estrogen receptor subtypes. Mooxifen is closely related to chemotherapy sensitivity. Tamoxifen has a synergistic killing effect with cisplatin in the high estrogen receptor beta and low alpha expression of esophageal adenocarcinoma cell lines. There is no effect in the high estrogen receptor alpha and low beta expression of the esophageal adenocarcinoma cell lines. This suggests that we have high estrogen receptor beta, low alpha expression of esophageal adenocarcinoma patients. We try to combine tamoxifen with cisplatin to improve the clinical efficacy.

【學(xué)位授予單位】：天津醫(yī)科大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2015
【分類號】：R735.1

【參考文獻】

相關(guān)期刊論文 前1條

1 An-Hui Wang;Yuan Liu;Bo Wang;Yi-Xuan He;Ye-Xian Fang;Yong-Ping Yan;;Epidemiological studies of esophageal cancer in the era of genome-wide association studies[J];World Journal of Gastrointestinal Pathophysiology;2014年03期

，

本文編號：1789711