本文選題：外泌體 + 宮頸癌��； 參考：《現(xiàn)代婦產(chǎn)科進(jìn)展》2017年03期

【摘要】：目的:研究宮頸癌細(xì)胞Siha分泌的外泌體(Exosomes)能否介導(dǎo)上皮-間質(zhì)轉(zhuǎn)化(EMT),從而改變癌前細(xì)胞Ect1的體外侵襲能力。方法:差速超速離心法分離SihaExo,透射電鏡觀察形態(tài),Western blot檢測(cè)標(biāo)志蛋白表達(dá),共聚焦顯微鏡觀察PKH67染色外泌體的生物學(xué)活性。Siha-Exo和PBS分別與Ect1共培養(yǎng)48h后Western blot檢測(cè)上皮和間質(zhì)轉(zhuǎn)化標(biāo)志蛋白的表達(dá),Transwell侵襲實(shí)驗(yàn)檢測(cè)共培養(yǎng)后Ect1體外侵襲能力。結(jié)果:Siha-Exo為直徑30~100nm的盤(pán)狀囊泡,特異性表達(dá)標(biāo)志性蛋白CD63和CD81,能夠被受體細(xì)胞Ect1內(nèi)吞,具備生物學(xué)活性。Western blot結(jié)果顯示,與PBS組相比,Siha-Exo組下調(diào)Ect1細(xì)胞中上皮標(biāo)志蛋白E-Cadherin和β-Catenin的表達(dá),上調(diào)間質(zhì)標(biāo)志蛋白NCadherin和Vimentin的表達(dá)。Transwell侵襲實(shí)驗(yàn)顯示,與PBS組相比,Siha-Exo組穿透聚碳酯膜的Ect1細(xì)胞數(shù)明顯增多(P=0.01)。結(jié)論:Siha-Exo能夠介導(dǎo)Ect1發(fā)生EMT變化,提高其體外侵襲能力。
[Abstract]:Aim: to investigate whether exocrine exosomes secreted by cervical cancer cells (Siha) can mediate epithelial-interstitial transformation (EMT) and thus change the invasiveness of precancerous Ect1 in vitro. Methods: Siha Exo was isolated by differential ultracentrifugation, and the expression of marker protein was detected by Western blot. Confocal microscopy was used to observe the biological activity of exosomes by PKH67 staining. Siha-Exo and PBS were co-cultured with Ect1 for 48 h. Western blot was used to detect the expression of epithelial and interstitial transforming marker protein. Transwell invasion assay was used to detect the invasiveness of Ect1 after co-culture. Results: Siha-Exo was a disk-shaped vesicle with diameters of 30~100nm, which specifically expressed CD63 and CD81, and could be ingested by receptor cell Ect1. Western blot showed that the expression of E-Cadherin and 尾 -Catenin in Ect1 cells was down-regulated in Siha-Exo group compared with PBS group. Transwell invasion assay showed that compared with PBS group, the number of Ect1 cells in Siha-Exo group penetrating through polycarboxylate membrane was significantly higher than that in PBS group, and the number of Ect1 cells in Siha-Exo group was significantly higher than that in PBS group. Conclusion: Siha-Exo can mediate the change of EMT in Ect1 and improve its invasiveness in vitro.
【作者單位】： 南方醫(yī)科大學(xué)南方醫(yī)院婦產(chǎn)科;南方醫(yī)科大學(xué)南方醫(yī)院病理科;
【基金】：國(guó)家自然科學(xué)基金(No:81072132;81372871;81672589;JCYJ20160429161218745)
【分類號(hào)】：R737.33

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