本文選題：核糖體蛋白S激酶 + 宮頸癌�。� 參考：《中國(guó)老年學(xué)雜志》2017年19期

【摘要】：目的探討RNA干擾核糖體蛋白S6激酶1(S6K1)對(duì)人宮頸癌細(xì)胞Hela侵襲遷移及相關(guān)基因表達(dá)的影響。方法根據(jù)文獻(xiàn)報(bào)道及siRNA設(shè)計(jì)原則合成靶向S6K1基因的siRNA,以陽(yáng)離子脂質(zhì)體Lipofectamine 2000為載體轉(zhuǎn)染對(duì)數(shù)生長(zhǎng)期Hela細(xì)胞(沉默組),同時(shí)設(shè)轉(zhuǎn)染不針對(duì)任何基因隨機(jī)序列的陰性對(duì)照組和轉(zhuǎn)染試劑的空白對(duì)照組。轉(zhuǎn)染48 h后采用實(shí)時(shí)定量聚合酶鏈反應(yīng)(qRT-PCR)檢測(cè)干擾效率,分別于轉(zhuǎn)染24、48、72 h后采用WST法評(píng)價(jià)靶向沉默S6K1對(duì)Hela細(xì)胞增殖的影響,采用Transwell小室法檢測(cè)經(jīng)S6K1沉默后對(duì)Hela細(xì)胞侵襲遷移的影響,采用Western印跡法檢測(cè)S6K1沉默后Hela細(xì)胞中人基質(zhì)金屬蛋白酶(MMP)-2、MMP-9和基質(zhì)金屬蛋白酶組織抑制因子(TIMP)-1、TIMP-2的蛋白水平。結(jié)果以空白對(duì)照組為參照(其S6K1 mRNA水平為1.000),陰性對(duì)照組的S6K1 mRNA水平為(1.027±0.034),差異無(wú)統(tǒng)計(jì)學(xué)意義(P0.05),而沉默組的S6K1 mRNA水平(0.158±0.012)低于其余兩組(P0.05),相對(duì)于空白對(duì)照組的沉默率為(92.4±3.7)%。與空白對(duì)照組和陰性對(duì)照組比較,沉默組轉(zhuǎn)染后的增殖率降低,遷移實(shí)驗(yàn)和侵襲實(shí)驗(yàn)中的穿膜細(xì)胞數(shù)均降低,MMP-2、MMP-9蛋白水平均降低,而TIMP-1、TIMP-2蛋白水平均升高(P0.05)。結(jié)論 RNA靶向沉默S6K1表達(dá)可抑制人宮頸癌Hela細(xì)胞侵襲遷移,可能與其抑制MMP-2/9表達(dá)及促進(jìn)TIMP-1/2表達(dá)有關(guān)。
[Abstract]:Objective to investigate the effects of RNA interfering ribosomal protein S6 kinase 1 (S6K1) on Hela invasion and migration and related gene expression in human cervical cancer cells. Methods siRNAs targeting S6K1 gene were synthesized according to literature reports and siRNA design principles. Cationic liposome Lipofectamine 2000 was used as vector to transfect logarithmic growth phase Hela cells (silencing group). Irradiation group and blank control group of transfection reagent. The interference efficiency was detected by real-time quantitative polymerase chain reaction qRT-PCR 48 h after transfection, and the effect of targeted silencing S6K1 on the proliferation of Hela cells was evaluated by WST assay after transfection for 48 h. Transwell chamber assay was used to detect the effect of S6K1 silencing on the invasion and migration of Hela cells, and Western blot was used to detect the protein levels of human matrix metalloproteinase MMP-2MMP-2MMP-9 and tissue inhibitor of matrix metalloproteinase-1 (TIMP-1) in Hela cells after S6K1 silencing. Results compared with the control group (the S6K1 mRNA level was 1.000g), the S6K1 mRNA level of the negative control group was 1.027 鹵0.034 (P 0.05), but the S6K1 mRNA level of the silencing group (0.158 鹵0.012) was lower than that of the other two groups (P 0.05), and the silencing rate was 92.4 鹵3.7 in comparison with the blank control group. Compared with the blank control group and the negative control group, the proliferation rate of the silencing group decreased, and the number of transmembrane cells in migration and invasion experiments decreased, while the level of TIMP-1 and TIMP-2 protein increased (P 0.05). Conclusion RNA targeting silencing of S6K1 expression can inhibit the invasion and migration of human cervical cancer Hela cells, which may be related to the inhibition of MMP-2/9 expression and the promotion of TIMP-1/2 expression.
【作者單位】： 承德醫(yī)學(xué)院附屬醫(yī)院婦科;南京醫(yī)科大學(xué)附屬南京市婦幼保健院醫(yī)學(xué)研究中心;
【基金】：國(guó)家自然科學(xué)基金(81302304)
【分類(lèi)號(hào)】：R737.33

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