摘要
我們已經(jīng)開(kāi)發(fā)出一種通過(guò)定位pH敏感的熒光染料檢測(cè)細(xì)胞表面的pH值，seminaphtharhodafluor（78）， 結(jié)合pH低插入肽（Phlip。據(jù)觀察，許多病變組織是酸性的，腫瘤 尤其如此。結(jié)合影響腫瘤細(xì)胞內(nèi)酸化，細(xì)胞泵出乳酸和質(zhì)子 維持細(xì)胞內(nèi)pH值，酸化細(xì)胞外空間。碳酸酐酶對(duì)細(xì)胞表達(dá)的表面有助于進(jìn)一步 酸化。因此，腫瘤細(xì)胞表面附近的pH值是預(yù)期要低，隨著距離 膜，所以體積測(cè)量pH值不會(huì)報(bào)告表面酸度。我們的新的表面pH值的測(cè)量工具是 癌細(xì)胞生長(zhǎng)在球體的驗(yàn)證，在小鼠體內(nèi)腫瘤模型，在切除腫瘤。我們發(fā)現(xiàn)，表面的pH敏感 細(xì)胞糖酵解活動(dòng)：pH降低高血糖和增加如果葡萄糖與nonmetabolized脫氧葡萄糖代替。 高轉(zhuǎn)移性癌細(xì)胞，pH測(cè)量表面6.7–6.8，當(dāng)外部的pH為7.4。 的足夠敏感單元在腫瘤誘導(dǎo)腹腔注射在體內(nèi)的變化來(lái)檢測(cè)0.2–0.3 pH值方法的葡萄糖。 在表面的pH高轉(zhuǎn)移性的腫瘤細(xì)胞內(nèi)發(fā)現(xiàn)約6.1–6.4，而在非轉(zhuǎn)移性腫瘤， 是6.7–6.9，可能創(chuàng)造更積極的方式來(lái)區(qū)分。其他生物的作用 表面酸度可能會(huì)發(fā)現(xiàn)，現(xiàn)在有針對(duì)性的測(cè)量是可能的。
本文由專(zhuān)注于提供生物科技服務(wù)的ELISA試劑盒廠(chǎng)商齊一生物收集整理
原文：
AbstractWe have developed a way to measure cell surface pH by positioning a pH-sensitive fluorescent dye, seminaphtharhodafluor (SNARF), conjugated to the pH low insertion peptide (pHLIP). It has been observed that many diseased tissues are acidic and that tumors are especially so. A combination of effects acidifies tumor cell interiors, and cells pump out lactic acid and protons to maintain intracellular pH, acidifying the extracellular space. Overexpression of carbonic anhydrases on cell surfaces further contributes to acidification. Thus, the pH near tumor cell surfaces is expected to be low and to increase with distance from the membrane, so bulk pH measurements will not report surface acidity. Our new surface pH-measurement tool was validated in cancer cells grown in spheroids, in mouse tumor models in vivo, and in excised tumors. We found that the surface pH is sensitive to cell glycolytic activity: the pH decreases in high glucose and increases if glucose is replaced with nonmetabolized deoxyglucose. For highly metastatic cancer cells, the pH measured at the surface was 6.7–6.8, when the surrounding external pH was 7.4. The approach is sensitive enough to detect 0.2–0.3 pH unit changes in vivo in tumors induced by i.p. injection of glucose. The pH at the surfaces of highly metastatic cells within tumors was found to be about 6.1–6.4, whereas in nonmetastatic tumors, it was 6.7–6.9, possibly creating a way to distinguish more aggressive from less aggressive tumors. Other biological roles of surface acidity may be found, now that targeted measurements are possible.
本文由專(zhuān)注于提供生物科技服務(wù)的ELISA試劑盒廠(chǎng)商齊一生物收集整理
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