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慢病毒載體介導(dǎo)的miR-126穩(wěn)定高表達(dá)促進(jìn)胃癌AGS細(xì)胞的遷移和侵襲

發(fā)布時(shí)間:2018-03-14 06:04

  本文選題:胃腫瘤 切入點(diǎn):miR- 出處:《實(shí)用醫(yī)學(xué)雜志》2017年05期  論文類型:期刊論文


【摘要】:目的:構(gòu)建穩(wěn)定高表達(dá)miR-126的胃癌AGS重組細(xì)胞系,并研究miR-126在體外對(duì)該胃癌細(xì)胞系增殖、轉(zhuǎn)移能力的影響。方法:構(gòu)建慢病毒Lv-has-miR-126感染AGS細(xì)胞系,經(jīng)qRT-PCR檢測(cè)miR-126表達(dá)差異,確認(rèn)miR-126穩(wěn)定高表達(dá)重組細(xì)胞系構(gòu)建成功后,應(yīng)用CCK-8和平板克隆形成實(shí)驗(yàn)檢測(cè)細(xì)胞體外增殖能力變化;Transwell遷移及侵襲實(shí)驗(yàn)檢測(cè)細(xì)胞在體外的遷移、侵襲能力。結(jié)果:重組細(xì)胞系構(gòu)建成功,綠色熒光表達(dá)良好,qRT-PCR證實(shí)重組細(xì)胞miR-126的表達(dá)水平較對(duì)照細(xì)胞顯著增高(P0.05)。CCK-8增殖及克隆形成實(shí)驗(yàn)顯示上調(diào)miR-126后胃癌細(xì)胞生長(zhǎng)及克隆形成數(shù)無明顯差異;而在遷移及侵襲實(shí)驗(yàn)中miR-126高表達(dá)組細(xì)胞遷移及侵襲能力顯著高于對(duì)照組(P0.05)。結(jié)論:成功構(gòu)建了穩(wěn)定高表達(dá)miR-126的AGS重組細(xì)胞系,初步證實(shí)了miR-126具有促進(jìn)胃癌AGS細(xì)胞遷移與侵襲的作用,提示miR-126可能與胃癌的轉(zhuǎn)移密切相關(guān)。
[Abstract]:Objective: to construct a recombinant AGS cell line with stable and high expression of miR-126, and to study the effect of miR-126 on the proliferation and metastasis of gastric cancer cell line in vitro. Methods: AGS cell line infected with lentivirus Lv-has-miR-126 was constructed and the difference of miR-126 expression was detected by qRT-PCR. After the successful construction of the recombinant cell line with stable and high expression of miR-126, the changes of cell proliferation ability in vitro were detected by CCK-8 and plate clone formation assay and the migration in vitro was detected by transwell migration assay and invasion assay, respectively. Invasiveness. Results: recombinant cell lines were successfully constructed, The expression level of miR-126 in recombinant cells was significantly higher than that in control cells by green fluorescence RT-PCR. The results showed that there was no significant difference in the growth and clone formation of gastric cancer cells after up-regulation of miR-126. The migration and invasion ability of the cells in the high expression group of miR-126 was significantly higher than that in the control group (P 0.05). Conclusion: the recombinant AGS cell line with stable and high expression of miR-126 was successfully constructed. It is preliminarily confirmed that miR-126 can promote the migration and invasion of gastric cancer AGS cells, suggesting that miR-126 may be closely related to gastric cancer metastasis.
【作者單位】: 南方醫(yī)科大學(xué)南方醫(yī)院檢驗(yàn)科;
【基金】:廣東省自然基金項(xiàng)目(編號(hào):2016A030313525) 廣州市科技計(jì)劃項(xiàng)目(編號(hào):201607010015)
【分類號(hào)】:R735.2

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