印跡基因CDKN1C對(duì)滋養(yǎng)細(xì)胞生物學(xué)功能的影響
發(fā)布時(shí)間:2018-02-22 00:39
本文關(guān)鍵詞: 印跡基因 CDKNC 滋養(yǎng)細(xì)胞 出處:《廣東醫(yī)學(xué)》2017年07期 論文類型:期刊論文
【摘要】:目的研究印跡基因CDKN1C對(duì)人滋養(yǎng)細(xì)胞生物學(xué)功能的影響。方法利用小干擾RNA沉默CDKN1C基因的表達(dá),將人絨毛外滋養(yǎng)細(xì)胞(TEV-1細(xì)胞株)分為3組,空白組細(xì)胞未經(jīng)轉(zhuǎn)染處理,對(duì)照組細(xì)胞由不含siRNA的空白脂粒體進(jìn)行轉(zhuǎn)染,siRNA組細(xì)胞由包裹siRNA的脂粒體進(jìn)行轉(zhuǎn)染。實(shí)時(shí)熒光定量-PCR(qRTPCR)和免疫印跡(Western blot)檢測(cè)TEV-1細(xì)胞CDKN1C基因的表達(dá)水平,CCK-8檢測(cè)細(xì)胞增殖率,流式細(xì)胞術(shù)檢測(cè)細(xì)胞凋亡及細(xì)胞周期,Transwell細(xì)胞遷移實(shí)驗(yàn)檢測(cè)細(xì)胞侵襲和遷移能力。結(jié)果 CDKN1C基因沉默后48h TEV-1細(xì)胞早期和晚期凋亡率均降低,細(xì)胞增殖率升高;細(xì)胞周期G0/G1期比例下降、S期和G2/M期比例增高;細(xì)胞遷移和侵襲能力增強(qiáng)(P0.05)。結(jié)論印跡基因CDKN1C對(duì)人滋養(yǎng)細(xì)胞增殖、凋亡、細(xì)胞周期、遷移、侵襲等生物學(xué)功能有調(diào)控作用。
[Abstract]:Objective to study the effect of imprinted gene CDKN1C on the biological function of human trophoblastic cells. Methods Human extracellular trophoblast cell line (TEV-1 cell line) was divided into three groups by silencing the expression of CDKN1C gene by small interfering RNA. The cells in the blank group were not treated with transfection. In the control group, the cells were transfected by lipofectin coated with siRNA. The expression level of CDKN1C gene in TEV-1 cells was detected by real-time fluorescence quantification-PCRnqRTPCRand Western blotting. CCK-8 was used to detect the proliferation rate of TEV-1 cells. Flow cytometry was used to detect cell apoptosis and cell cycle transwell cell migration. Results the early and late apoptosis rate of TEV-1 cells was decreased and the cell proliferation rate was increased 48 hours after CDKN1C gene silencing. The proportion of G _ 0 / G _ 1 phase decreased in S phase and G _ 2 / M phase, and the cell migration and invasion increased (P 0.05). Conclusion the imprinted gene CDKN1C can regulate the biological functions of human trophoblastic cell proliferation, apoptosis, cell cycle, migration and invasion.
【作者單位】: 廣州醫(yī)科大學(xué)附屬第二醫(yī)院產(chǎn)科;中山大學(xué)附屬第一醫(yī)院口腔科;廣東省婦幼保健院醫(yī)學(xué)遺傳中心;廣東省人民醫(yī)院婦產(chǎn)科;中山大學(xué)附屬第一醫(yī)院胎兒醫(yī)學(xué)中心;
【基金】:廣東省自然科學(xué)基金資助項(xiàng)目(編號(hào):2016A030310176) 廣州市科技計(jì)劃項(xiàng)目(編號(hào):1563000549) 廣州醫(yī)科大學(xué)博士啟動(dòng)基金項(xiàng)目(編號(hào):2015C16)
【分類號(hào)】:R329.2
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