【摘要】：目的探討腺相關(guān)病毒介導(dǎo)的CDK2-shRNA對(duì)人黑色素瘤細(xì)胞A375凋亡,為黑色素瘤的防治提供技術(shù)支持。方法體外合成CDK2及轉(zhuǎn)錄終止序列,構(gòu)建CDK2-shRNA的腺相關(guān)病毒載體和包裝相應(yīng)的重組腺相關(guān)病毒,熒光顯微鏡檢測(cè)轉(zhuǎn)染和感染條件下CDK2-shRNA表達(dá)效果,MTT法檢測(cè)細(xì)胞活力,Annexin V-FITC/PI細(xì)胞凋亡試劑盒檢測(cè)細(xì)胞凋亡,免疫印跡檢測(cè)cyclin E和E2F1表達(dá)。結(jié)果轉(zhuǎn)染和感染的腺相關(guān)病毒介導(dǎo)的CDK2-shRNA在A375細(xì)胞均有表達(dá)且具有較低的細(xì)胞毒性;轉(zhuǎn)染和感染的CDK2-shRNA均促進(jìn)A375細(xì)胞發(fā)生凋亡;干擾CDK2可降低cyclin E和E2F1表達(dá)。結(jié)論腺相關(guān)病毒介導(dǎo)的CDK2-shRNA促進(jìn)黑色素瘤細(xì)胞A375凋亡。
[Abstract]:Objective to investigate the apoptosis of human melanoma cell line A375 mediated by adeno-associated virus (CDK2-shRNA), and to provide technical support for the prevention and treatment of melanoma. Methods CDK2 and transcriptional termination sequence were synthesized in vitro. The adeno-associated virus vector of CDK2-shRNA and the corresponding recombinant adeno-associated virus were constructed. The expression of CDK2-shRNA was detected by fluorescence microscope, the cell vitality was detected by MTT assay, the apoptosis was detected by Annexin V-FITC/PI apoptosis kit, and the expression of cyclin E and E2F1 was detected by immunoblotting. Results both transfected and infected adeno-associated virus-mediated CDK2-shRNA were expressed in A375 cells and had low cytotoxicity. Both transfected and infected CDK2-shRNA promoted apoptosis of A375 cells. Interference with CDK2 could decrease the expression of cyclin E and E2F1. Conclusion adeno-associated virus mediated CDK2-shRNA promotes apoptosis of melanoma cell line A375.
【作者單位】： 黑龍江中醫(yī)藥大學(xué);福建中醫(yī)藥大學(xué)附屬廈門第三醫(yī)院皮膚科;吉林大學(xué)中日聯(lián)誼醫(yī)院普通外科;
【基金】：廈門市衛(wèi)生局資助項(xiàng)目(3502Z20130190) 廈門市同安區(qū)科技局資助項(xiàng)目
【分類號(hào)】：R739.5

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