本文關(guān)鍵詞： 細(xì)胞遺傳學(xué) Snail 成纖維細(xì)胞 乳腺癌 耐藥　出處：《中國科技論文》2017年18期 　論文類型：期刊論文

【摘要】：構(gòu)建穩(wěn)定表達(dá)Snail的成纖維細(xì)胞株并檢測其對乳腺癌細(xì)胞耐藥的影響,運(yùn)用pL-tdTomato-Neo和pL-tdTomato-mSnail轉(zhuǎn)染小鼠成纖維細(xì)胞3T3和L929,G418富集,單克隆挑選穩(wěn)定細(xì)胞株。采用實(shí)時熒光定量PCR和Western blotting檢測細(xì)胞內(nèi)Snail的表達(dá)。將Snail穩(wěn)定表達(dá)的成纖維細(xì)胞的培養(yǎng)上清作為條件培養(yǎng)基,按30%和60%的比例與乳腺癌細(xì)胞4T1共培養(yǎng),噻唑藍(lán)(MTT)檢測4T1對順鉑和阿霉素的IC_(50)。運(yùn)用實(shí)時熒光定量PCR檢測過表達(dá)Snail的成纖維中各種細(xì)胞因子的表達(dá)。結(jié)果表明:穩(wěn)定轉(zhuǎn)染細(xì)胞株胞體發(fā)出紅色熒光,胞內(nèi)Snail水平上調(diào),成功構(gòu)建了穩(wěn)定表達(dá)Snail的成纖維細(xì)胞株。MTT實(shí)驗發(fā)現(xiàn),4T1與過表達(dá)Snail的成纖維細(xì)胞共培養(yǎng)后,對順鉑和阿霉素的IC_(50)均升高。實(shí)時熒光定量PCR實(shí)驗發(fā)現(xiàn),Snail能誘導(dǎo)成纖維細(xì)胞多種細(xì)胞因子的表達(dá)增加。在成纖維細(xì)胞中,Snail可能通過調(diào)控多種細(xì)胞因子的表達(dá)參與乳腺癌細(xì)胞對化學(xué)藥物的抵抗。
[Abstract]:A fibroblast cell line expressing Snail stably was constructed and its effect on drug resistance of breast cancer cells was detected. Mouse fibroblasts 3T3 and L929 were transfected with pL-tdTomato-Neo and pL-tdTomato-mSnail and enriched with G418. Monoclonal selection of stable cell lines. Real-time fluorescence quantitative PCR and Western. Blotting was used to detect the expression of Snail. The culture supernatant of fibroblasts with stable expression of Snail was used as the conditioned medium. Breast cancer cells 4T1 were co-cultured in the proportion of 30% and 60%. Detection of 4T1 for cisplatin and adriamycin by MTT). The expression of cytokines in fibroblasts expressing Snail was detected by real-time fluorescence quantitative PCR. The results showed that the cell bodies of stable transfected cells emit red fluorescence. The level of intracellular Snail was up-regulated. A stable fibroblast cell line expressing Snail was successfully constructed. The expression of multiple cytokines in fibroblasts was increased by real-time fluorescence quantitative PCR assay. Snail may be involved in the resistance of breast cancer cells to chemical drugs by regulating the expression of many cytokines.
【作者單位】： 中山大學(xué)藥學(xué)院;
【基金】：高等學(xué)校博士學(xué)科點(diǎn)專項科研基金資助項目(20130171110056) 國家自然科學(xué)基金資助項目(81672943,81472643)
【分類號】：R737.9
【正文快照】： 乳腺癌是全球女性發(fā)病率最高的惡性腫瘤,也是我國發(fā)病率上升最快的惡性腫瘤之一[1]。然而,腫瘤耐藥往往導(dǎo)致乳腺癌化療失敗[2-3],腫瘤耐藥的機(jī)制及其分子調(diào)控網(wǎng)絡(luò)并未闡明,繼續(xù)探索腫瘤耐藥的機(jī)制為提高化療療效、攻克腫瘤提供理論依據(jù)。腫瘤微環(huán)境主要由大量的異質(zhì)性間質(zhì)細(xì)胞

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