microRNA-10b與胰腺癌吉西他濱耐藥的相關(guān)性及機(jī)制研究
發(fā)布時(shí)間:2018-07-03 16:07
本文選題:microRNA-b + 胰腺癌 ; 參考:《南京醫(yī)科大學(xué)學(xué)報(bào)(自然科學(xué)版)》2017年07期
【摘要】:目的:探討microRNA-10b(miR-10b)與胰腺癌細(xì)胞吉西他濱(gemcitabine,GEM)化療耐藥的相關(guān)性及可能機(jī)制。方法:RT-qPCR檢測吉西他濱相對耐藥的胰腺癌細(xì)胞株P(guān)ANC-1及吉西他濱相對敏感的CFPAC-1中miR-10b的表達(dá)情況;用不同濃度的吉西他濱作用于上述細(xì)胞,RT-qPCR檢測二者miR-10b的表達(dá)變化;CFPAC-1細(xì)胞轉(zhuǎn)染miR-10b mimics上調(diào)miR-10b表達(dá)量,CCK-8法及流式細(xì)胞儀檢測細(xì)胞對吉西他濱藥物敏感性的變化,Western blot檢測抗凋亡相關(guān)蛋白(如PI3K、p-Akt、Bcl-2、Survivin)的表達(dá),RT-qPCR檢測PTEN基因的表達(dá)變化。結(jié)果:PANC-1細(xì)胞中miR-10b的表達(dá)顯著高于CFPAC-1細(xì)胞,且上述兩種細(xì)胞中miR-10b的表達(dá)量與吉西他濱呈濃度梯度依賴;高表達(dá)miR-10b后CFPAC-1細(xì)胞對吉西他濱的敏感性下降,PI3K、p-Akt、Bcl-2、Survivin蛋白的表達(dá)升高,PTEN mRNA的表達(dá)水平降低。結(jié)論:miR-10b可能通過負(fù)性調(diào)控PTEN的表達(dá)水平,從而增加PI3K、p-Akt、Bcl-2、Survivin蛋白的表達(dá),減少凋亡來降低CFPAC-1對吉西他濱的敏感性,最終導(dǎo)致胰腺癌細(xì)胞對吉西他濱化療耐受。
[Abstract]:Aim: to investigate the relationship between microRNA-10b (miR-10b) and chemotherapeutic resistance of pancreatic cancer cell gemcitabine gem and its possible mechanism. Methods the expression of miR-10b in the relatively resistant pancreatic cancer cell line PANC-1 and the relatively sensitive CFPAC-1 was detected by RT-qPCR. Expression of miR-10b in the above cells treated with different concentrations of gemcitabine was detected by RT-qPCR. The expression of miR-10b mimics was up-regulated by CFPAC-1 cells. CCK-8 assay and flow cytometry were used to detect the sensitivity of the cells to gemcitabine. Western blot was used to detect the expression of miR-10b in CFPAC-1 cells. The expression of anti-apoptosis-related proteins (such as PI3KUp-AktC2Bcl-2survivin) was detected by RT-qPCR to detect the expression of PTEN gene. Results the expression of miR-10b in the cell line was significantly higher than that in the CFPAC-1 cell line, and the expression of miR-10b in the two cells was in a concentration-dependent manner. After overexpression of miR-10b, the sensitivity of CFPAC-1 cells to gemcitabine was decreased. Conclusion the expression of PTEN may be negatively regulated by 1: miR-10b, which may increase the expression of PI3KP- Aktnbcl-2survivin protein, reduce apoptosis and decrease the sensitivity of CFPAC-1 to gemcitabine, leading to the chemotherapeutic tolerance of pancreatic cancer cells to gemcitabine.
【作者單位】: 南京醫(yī)科大學(xué)附屬鼓樓臨床醫(yī)學(xué)院;啟東市人民醫(yī)院消化內(nèi)科;南京大學(xué)醫(yī)學(xué)院附屬鼓樓醫(yī)院消化科;
【基金】:[基金項(xiàng)目]江蘇省醫(yī)學(xué)領(lǐng)軍人才與創(chuàng)新團(tuán)隊(duì)(LJ201104)
【分類號】:R735.9
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