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siRNA干擾KLF8表達(dá)對(duì)鼻咽癌細(xì)胞上皮間質(zhì)轉(zhuǎn)化的作用

發(fā)布時(shí)間:2019-06-11 05:54
【摘要】:目的探討下調(diào)KLF8的表達(dá)對(duì)鼻咽癌CNE1-LMP1細(xì)胞侵襲能力的影響及其機(jī)制。方法通過(guò)脂質(zhì)體轉(zhuǎn)染法將KLF8 siRNA真核表達(dá)質(zhì)粒穩(wěn)定轉(zhuǎn)染至鼻咽癌細(xì)胞株CNE1-LMP1。Western blot和RT-PCR法檢測(cè)CNE1-LMP1細(xì)胞中KLF8、E-cadherin、N-cadherin蛋白和m RNA表達(dá)水平的變化。Transwell侵襲實(shí)驗(yàn)觀察siRNA后對(duì)CNE1-LMP1細(xì)胞侵襲能力的影響。結(jié)果 KLF8 siRNA轉(zhuǎn)染的CNE1-LMP1細(xì)胞株與其陰性對(duì)照NC-si組比較KLF8蛋白和mRNA表達(dá)水平均下調(diào),證實(shí)穩(wěn)轉(zhuǎn)細(xì)胞株建立成功。KLF8 siRNA可上調(diào)CNE1-LMP1細(xì)胞中E-cadherin的表達(dá),而下調(diào)N-cadherin的表達(dá)。Transwell侵襲實(shí)驗(yàn)顯示siRNA干擾CNE1-LMP1細(xì)胞侵襲能力下降。結(jié)論 KLF8在鼻咽癌CNE1-LMP1細(xì)胞中高表達(dá);通過(guò)siRNA下調(diào)KLF8的表達(dá)能干擾上皮間質(zhì)轉(zhuǎn)化(EMT)相關(guān)蛋白E-cadherin、N-cadherin的表達(dá),阻斷EMT過(guò)程,抑制CNE1-LMP1細(xì)胞侵襲轉(zhuǎn)移。
[Abstract]:Objective to investigate the effect of down-regulating the expression of KLF8 on the invasiveness of nasopharyngeal carcinoma (NPC) CNE1-LMP1 cells and its mechanism. Methods KLF8 siRNA eukaryotic expression plasmid was stably transferred into nasopharyngeal carcinoma cell line CNE1-LMP1.Western blot and RT-PCR by liposomes transfer method to detect KLF8,E-cadherin, in CNE1-LMP1 cells. The expression levels of N-cadherin protein and m RNA were changed. Transwell invasion assay was used to observe the effect of siRNA on the invasiveness of CNE1-LMP1 cells. Results the expression of KLF8 protein and mRNA in CNE1-LMP1 cell line transfected with KLF8 siRNA was down-regulated compared with that in negative control NC-si group, which confirmed that the stable cell line was established successfully. KLF 8 siRNA could up-regulate the expression of E-cadherin in CNE1-LMP1 cells. The expression of N-cadherin was down-regulated. Transwell invasion test showed that siRNA interfered with the invasive ability of CNE1-LMP1 cells. Conclusion the expression of KLF8 is highly expressed in nasopharyngeal carcinoma CNE1-LMP1 cells, and the down-regulation of KLF8 expression by siRNA can interfere with the expression of (EMT)-associated protein E 鈮,

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