【摘要】：【目的】對(duì)紫色桿菌素作用后的結(jié)腸癌細(xì)胞HT29進(jìn)行差異蛋白質(zhì)組學(xué)分析,探究其影響的代謝通路,揭示其抑制癌細(xì)胞生長(zhǎng)的作用機(jī)制,為新型抗癌藥物的開發(fā)提供一定的參考。【方法】以不同劑量的紫色桿菌素處理HT29 24、48、72 h,通過MTT試驗(yàn)以及透射電鏡分析其對(duì)結(jié)腸癌細(xì)胞的有效抑制劑量及抑制情況。在此基礎(chǔ)上,對(duì)提取的3個(gè)處理組的蛋白進(jìn)行同位素標(biāo)記,利用反相液相色譜(RP-LC)聯(lián)用串聯(lián)質(zhì)譜(AB SCIEX Triple TOF5600)獲得樣品中的多肽及其相對(duì)豐度信息,通過數(shù)據(jù)庫(NCBI.Human.protein database)搜索比對(duì),鑒定差異表達(dá)蛋白。利用GO分析、KEGG信號(hào)通路分析,解析紫色桿菌素抑癌作用代謝途徑�！窘Y(jié)果】紫色桿菌素對(duì)HT29的抑制作用呈一定的時(shí)間、劑量依賴性。當(dāng)紫色桿菌素對(duì)HT29的抑制率達(dá)50%時(shí),僅為陽性藥物5-Fu劑量的1/6,具有更明顯的抑制效果。電鏡下觀察顯示,隨著紫色桿菌素劑量增大,細(xì)胞內(nèi)部線粒體出現(xiàn)空泡結(jié)構(gòu),質(zhì)膜出泡;當(dāng)濃度達(dá)30 mg·L~(-1)時(shí),細(xì)胞膜消失,染色質(zhì)邊集。通過差異蛋白質(zhì)組學(xué)分析,共鑒定出4 258個(gè)蛋白,表達(dá)差異在2倍以上的蛋白共為757個(gè)。其中高劑量組處理差異蛋白數(shù)為492個(gè),低劑量組處理的差異蛋白數(shù)為112個(gè),陽性對(duì)照組差異蛋白數(shù)為336個(gè)。KEGG分析顯示這些差異蛋白共參與50條信號(hào)通路。其中有10條信號(hào)通路具有顯著性(P0.05),主要參與核糖體循環(huán)途徑、三羧酸循環(huán)途徑和RNA降解途徑等�！窘Y(jié)論】紫色桿菌素主要通過影響HT29細(xì)胞生命活動(dòng)中的蛋白轉(zhuǎn)錄和翻譯水平進(jìn)而發(fā)揮抑制作用。
[Abstract]:[objective] to analyze the differential proteome of colon cancer cell line HT29 treated with purple baculins, to explore the metabolic pathway of its effect, to reveal the mechanism of its inhibitory effect on the growth of cancer cells, and to provide some reference for the development of new anticancer drugs. [methods] the effective inhibitory dose and inhibition of HT29 on colon cancer cells were analyzed by MTT test and transmission electron microscope. [methods] HT29 was treated with different doses of purple bacteriocin for 48 h, 72 h, and its effective inhibitory dose and inhibition on colon cancer cells were analyzed by MTT test and transmission electron microscope. On this basis, the proteins of the three treatment groups were labeled by isotope, and the polypeptide and its relative abundance information were obtained by reversed-phase liquid chromatography (RP-LC) coupled with tandem mass spectrometry (AB SCIEX Triple TOF5600). The differentially expressed proteins were identified by database (NCBI.Human.protein database) search and comparison. GO analysis and KEGG signal pathway analysis were used to analyze the metabolic pathway of purple baculins in inhibiting cancer. [results] the inhibitory effect of purple baculins on HT29 was time-dependent and dose-dependent. When the inhibition rate of purple baculins on HT29 was 50%, it was only 1 鈮，

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