本文選題：前列腺素E + 信號通路��； 參考：《中國老年學(xué)雜志》2017年15期

【摘要】：目的研究前列腺素(PGE2)激活Yes信號通路相關(guān)蛋白(YAP)正反饋回路促進(jìn)小鼠結(jié)腸炎后結(jié)腸再生及癌變的分子機(jī)制。方法將DLD1結(jié)腸癌細(xì)胞株或基因穩(wěn)定敲除細(xì)胞株,在有/無信號通路特異性抑制劑的條件下,采用重組PGE2或PBS孵育,采用免疫印跡法、免疫熒光法、定量PCR、轉(zhuǎn)錄和增殖法進(jìn)行比較分析。采用DSS法誘導(dǎo)C57/BL6小鼠結(jié)腸炎模型(對照鼠),同時(shí)構(gòu)建羥基前列腺素脫氫酶基因敲除鼠(15-PGFH-KO)、YAP基因敲除鼠(YAP-KO)、和雙基因敲除(DKO)小鼠,采用吲哚美辛抑制PGE2合成信號通路,檢測下游基因表達(dá)及結(jié)腸再生和癌變功能。結(jié)果 PGE2處理結(jié)腸癌細(xì)胞株導(dǎo)致YAP mRNA轉(zhuǎn)錄及蛋白表達(dá)水平升高,15-PGDH-KO組鼠結(jié)腸組織中PGE2的表達(dá)水平較對照敲除細(xì)胞(WT)組增長了2.5倍,吲哚美辛處理后15-PGDH-KO組鼠YAP的表達(dá)水平及下游靶基因水平均較WT鼠顯著升高。吲哚美辛處理顯著降低PG過氧化物合成酶(COX)-2及PGE受體4基因(EP4)的轉(zhuǎn)錄水平,轉(zhuǎn)染YAP持續(xù)激活突變體(5SA)可顯著增加COX-2和EP4的蛋白表達(dá)水平和轉(zhuǎn)錄水平,敲除YAP后上述現(xiàn)象消失。對照鼠口服葡聚糖硫酸鈉(DSS)導(dǎo)致結(jié)腸炎,注射PGE2促進(jìn)小鼠發(fā)生結(jié)腸再生;YAP敲除鼠并未發(fā)生結(jié)腸再生,同時(shí)口服DSS后迅速死亡;15-PGFH敲除鼠結(jié)腸組織再生比對照鼠更快,且體重恢復(fù)更快、結(jié)腸長度和結(jié)腸炎組織學(xué)得分更高。上述現(xiàn)象可以被注射吲哚美辛逆轉(zhuǎn)。YAP敲除鼠或15-PGFH敲除鼠誘導(dǎo)結(jié)腸炎后未發(fā)現(xiàn)自發(fā)性腫瘤,YAP/15-PGDH雙基因敲除鼠出現(xiàn)息肉并最終發(fā)展成原位癌�？诜胚崦佬量捎行Х乐闺p敲除鼠自發(fā)性腫瘤的形成。結(jié)論 PGE2可增加YAP的表達(dá)水平和轉(zhuǎn)錄活性,從而促進(jìn)COX-2及EP4的表達(dá)水平增加,這一信號回路可促進(jìn)擴(kuò)散的結(jié)腸癌細(xì)胞系增殖和結(jié)腸炎小鼠結(jié)腸組織再生,持續(xù)性激活進(jìn)一步可導(dǎo)致息肉和小鼠結(jié)腸腫瘤的形成。
[Abstract]:Aim to study the molecular mechanism of prostaglandin PGE2 (PGE2) activation of Yes signal pathway associated protein (YAP) positive feedback loop in promoting colon regeneration and carcinogenesis after colitis in mice. Methods DLD1 colon cancer cell line or gene stable knockout cell line was incubated with recombinant PGE2 or PBS in the presence or absence of signal pathway specific inhibitors. Quantitative PCR, transcription and proliferation were compared and analyzed. The colitis model of C57/BL6 mice was induced by DSS method (control mice). The hydroxyl prostaglandin dehydrogenase gene knockout mice, 15-PGFH-KOAP gene knockout mice, and double-gene knockout mice were constructed. Indomethacin was used to inhibit the PGE2 synthesis signal pathway. The downstream gene expression and colon regeneration and carcinogenesis were detected. Results the expression of PGE2 in colon tissue of 15-PGDH-KO group was increased by 2.5-fold compared with that of control group. After indomethacin treatment, the expression level of YAP and the downstream target gene level in 15-PGDH-KO group were significantly higher than those in WT group. Indomethacin treatment significantly reduced the transcription level of COX-2 and PGE receptor 4 gene (EP4). Transfection of YAP continuously activated mutants (p5SA4) significantly increased the protein expression and transcription level of COX-2 and EP4, which disappeared after YAP knockout. The colitis was induced by oral dextran sodium sulfate in control mice. The colonic regeneration was not observed in mice induced by PGE2 injection, and the colonic tissue regeneration of 15-PGFH knockout mice was faster than that of control mice after oral DSS. Weight recovered faster, colon length and colitis histological scores were higher. These phenomena can be reversed by injection of indomethacin. YAP knockout mice or 15-PGFH knockout mice induced colitis. Oral administration of indomethacin can effectively prevent the formation of spontaneous tumor in double knockout mice. Conclusion PGE2 can increase the expression level and transcription activity of YAP, thus promote the expression of COX-2 and EP4. This signal loop can promote the proliferation of diffuse colon cancer cell line and the regeneration of colonic tissue in colitis mice. Continuous activation may further lead to polyp and colon tumor formation in mice.
【作者單位】： 牡丹江醫(yī)學(xué)院第二附屬醫(yī)院消化內(nèi)科;
【分類號】：R574.62

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