本文選題：微小RNA- + 胃癌細(xì)胞株AGS��； 參考：《中國(guó)腫瘤生物治療雜志》2017年10期

【摘要】：目的:探討miRNA-24對(duì)胃癌AGS細(xì)胞增殖和凋亡的影響及其潛在的作用機(jī)制。方法:實(shí)時(shí)定量聚合酶鏈?zhǔn)椒磻?yīng)(q RT-PCR)檢測(cè)不同胃癌細(xì)胞株(AGS、MKN74、HGC27)和胃黏膜上皮GES-1細(xì)胞中miRNA-24的表達(dá)水平。建立miRNA-24過表達(dá)的AGS細(xì)胞株,采用CCK-8法檢測(cè)細(xì)胞增殖活力,流式細(xì)胞術(shù)檢測(cè)細(xì)胞周期和凋亡情況,Western blotting檢測(cè)細(xì)胞周期和凋亡相關(guān)cyclin D1、CDK2、Bcl-2、p-IκB-α/IκB-α和p-Rb/Rb蛋白的表達(dá)水平;雙熒光素酶報(bào)告基因分析法預(yù)測(cè)和驗(yàn)證miRNA-24可能的靶基因。結(jié)果:3株胃癌細(xì)胞中miRNA-24的表達(dá)均低于GES-1 cell。轉(zhuǎn)染miRNA-24 mimic(miR-24組)48 h后AGS細(xì)胞增殖活力顯著低于miR-NC組[(119.62±12.63)%vs(147.79±11.89)%,P0.05],并出現(xiàn)周期阻滯,且早期和晚期細(xì)胞凋亡率明顯較miR-NC組上升[早期凋亡率:(11.32±2.27)%vs(0.57±0.08)%;晚期凋亡率:(15.56±2.27)%vs(0.85±0.16)%,均P0.05]。轉(zhuǎn)染miRNA-24 mimic后,細(xì)胞中cyclin D1、CDK2、Bcl-2及p-Rb的蛋白表達(dá)水平均較miR-NC組顯著降低,p-IκB-α蛋白的表達(dá)水平較miR-NC組顯著上升。共轉(zhuǎn)染miRNA-24 mimic和miRNA-24可能作用靶點(diǎn)CARMA3基因過表達(dá)質(zhì)粒的miR-24+pc DNA-CARMA3組AGS細(xì)胞CARMA3蛋白表達(dá)較miR-24組明顯增加(1.74±0.09 vs 1.03±0.06,P0.05)。miR-24+pc DNA3-CARMA3組AGS細(xì)胞48 h增殖活力較miR-24組顯著升高[(137.85±15.34)%vs(102.31±11.23)%,P0.05];而miR-24+pc DNA3-CARMA3組AGS細(xì)胞早期凋亡率和晚期凋亡率均較miR-24組顯著降低[早期凋亡率:(4.24±0.56)%vs(11.32±2.27)%,P0.05;晚期凋亡率:(6.38±0.63)%vs(15.56±2.27)%,P0.05]。CARMA3過表達(dá)可部分逆轉(zhuǎn)miRNA-24對(duì)胃癌AGS細(xì)胞增殖及凋亡的作用。結(jié)論:miRNA-24可通過靶向CARMA3基因抑制胃癌細(xì)胞的增殖、促進(jìn)其凋亡。
[Abstract]:Aim: to investigate the effect of miRNA-24 on proliferation and apoptosis of gastric cancer AGS cells and its possible mechanism.Methods: Real-time quantitative polymerase chain reaction (RT-PCR) was used to detect the expression of miRNA-24 in different gastric cancer cell lines (AGSN MKN74, HGC27) and gastric epithelial GES-1 cells.The miRNA-24 overexpression AGS cell line was established. The proliferative activity was detected by CCK-8 assay, and the cell cycle and apoptosis were detected by flow cytometry. The expression levels of cyclin D1- CDK2pBcl-2mp-I 魏 B- 偽 / I 魏 B- 偽 and p-Rb/Rb protein were detected by flow cytometry (FCM).Double luciferase reporter gene analysis was used to predict and verify the possible target genes of miRNA-24.Results the expression of miRNA-24 in the 3 strains of gastric cancer was lower than that in GES-1 cells.The proliferative activity of AGS cells in miRNA-24 mimic(miR-24 group was significantly lower than that in miR-NC group 48 h after transfection [119.62 鹵12.63)%vs(147.79 鹵11.89 + P 0.05], and cell cycle arrest was found, and the early and late apoptosis rate was significantly higher than that in miR-NC group [early apoptosis rate: 11.32 鹵2.27)%vs(0.57 鹵0.08; late apoptosis rate: 15.56 鹵2.27)%vs(0.85 鹵0.16, P0.05].After transfection of miRNA-24 mimic, the expression levels of Bcl-2 and p-Rb in cyclin D1- CDK2 and p-Rb were significantly lower than those in miR-NC group. The expression level of p-I 魏 B- 偽 protein was significantly increased compared with miR-NC group.The expression of CARMA3 protein in AGS cells of miR-24 PC DNA-CARMA3 group was significantly higher than that of miR-24 group in miR-24 PC DNA-CARMA3 group (1.74 鹵0.09 vs 1.03 鹵0.06) P0.05N. MiR-24 PC DNA3-CARMA3 group was significantly higher than miR-24 group [137.85 鹵15.34)%vs(102.31 鹵11.23], and miR-24 PC DNA3-CARMA3 group was AGS significantly higher than miR-24 group.The early apoptosis rate and late apoptosis rate were significantly lower than those in miR-24 group [the early apoptosis rate was 4.24 鹵0.56)%vs(11.32 鹵2.27; the late apoptotic rate was 6.38 鹵0.63)%vs(15.56 鹵2.27P 0.05] .CARMA3 overexpression could partially reverse the effect of miRNA-24 on the proliferation and apoptosis of gastric cancer AGS cells.Conclusion: miRNA-24 can inhibit the proliferation and promote apoptosis of gastric cancer cells by targeting CARMA3 gene.
【作者單位】： 新鄉(xiāng)醫(yī)學(xué)院第一附屬醫(yī)院消化內(nèi)科;
【分類號(hào)】：R735.2

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本文編號(hào)：1770846