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  本文選題：HT細胞 + MDR�。� 參考：《基因組學與應用生物學》2017年04期

【摘要】：以人類急性早幼粒白血病HT9細胞為實驗對象,利用RNA干擾(RNA interference,RNAi)技術(shù)干擾MDR1基因表達,提高HT9細胞對紫杉醇的敏感性。構(gòu)建了3種靶向MDR1基因的重組干擾載體,穩(wěn)定轉(zhuǎn)染HT9細胞后,采用qRT-PCR和Western blotting方法,分別檢測MDR1基因的m RNA和蛋白表達水平,用MTT法檢測各轉(zhuǎn)染組細胞對紫杉醇的敏感性,流式細胞術(shù)檢測各組細胞藥物外排功能。結(jié)果顯示,成功構(gòu)建了3種靶向MDR1的重組干擾載體p3.1-1、p3.1-2和p3.1-3。3種重組干擾載體不同程度抑制HT9細胞中MDR1基因的表達,其中重組干擾載體p3.1-3對MDR1基因沉默效果最好,對m RNA和蛋白的沉默效率分別為73.80%和47.61%,與對照組相比,轉(zhuǎn)染p3.1-3重組干擾載體的細胞對紫杉醇的IC50由(1.26±0.17)μg/m L降至(0.46±0.07)μg/m L,逆轉(zhuǎn)率達到(63.48±5.91)%,并且藥物外排功能也明顯降低。以上實驗結(jié)果表明,3種靶向MDR1的重組干擾載體均能抑制MDR1基因表達,其中p3.1-3對MDR1基因干擾效果最好,并且能夠有效提高HT9細胞對紫杉醇的敏感性,降低細胞藥物外排功能。
[Abstract]:HT9 cells of human acute promyelocytic leukemia (AHL) were used to interfere with the expression of MDR1 gene by using RNA interference RNAi technique to enhance the sensitivity of HT9 cells to paclitaxel. Three kinds of recombinant interference vectors targeting MDR1 gene were constructed. After stable transfection of HT9 cells, the expression levels of m RNA and protein of MDR1 gene were detected by qRT-PCR and Western blotting methods, and the sensitivity of transfected cells to paclitaxel was detected by MTT assay. The efflux function of the cells was detected by flow cytometry. The results showed that three recombinant interference vectors p3.1-1p3.1-2 and p3.1-3.3 were successfully constructed to inhibit the expression of MDR1 gene in HT9 cells. Among them, p3.1-3 had the best silencing effect on MDR1 gene. The silencing efficiency of m RNA and protein were 73.80% and 47.61%, respectively. Compared with the control group, the IC50 of the cells transfected with p3.1-3 recombinant interference vector decreased from 1.26 鹵0.17 渭 g / mL to 0.46 鹵0.07 渭 g / mL, the reversal rate was 63.48 鹵5.91g / mL, and the efflux function of the drug was also significantly decreased. The results showed that all three recombinant interference vectors targeting MDR1 could inhibit the expression of MDR1 gene, among which p3.1-3 had the best effect on MDR1 gene interference, and could effectively enhance the sensitivity of HT9 cells to paclitaxel and reduce the drug efflux function of HT9 cells.
【作者單位】： 齊齊哈爾大學生命科學與農(nóng)林學院;
【基金】：黑龍江省自然科學基金(項目編號:C200624,C201241) 黑龍江省教育廳科學技術(shù)項目(項目編號:11511447,12511611);黑龍江省教育廳基本業(yè)務專項理工重點項目(項目編號:135109104) 齊齊哈爾大學2015年研究生創(chuàng)新科研項目(項目編號:YJSCX2015-024X)共同資助
【分類號】：R733.71

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